Adv Physiol Educ 38: 355–365, 2014;
doi:10.1152/advan.00125.2013.
Using stimulation of the diving reflex in humans to teach integrative
physiology
Julia K. Choate, Kate M. Denton, Roger G. Evans, and Yvonne Hodgson
Department of Physiology, School of Biomedical Sciences, Monash University, Melbourne, Victoria, Australia
Submitted 4 November 2013; accepted in final form 5 September 2014
Choate JK, Denton KM, Evans RG, Hodgson Y. Using stimulation
of the diving reflex in humans to teach integrative physiology. Adv
Physiol Educ 38: 355–365, 2014; doi:10.1152/advan.00125.2013.—
During underwater submersion, the body responds by conserving O2
and prioritizing blood flow to the brain and heart. These physiological
adjustments, which involve the nervous, cardiovascular, and respira-
tory systems, are known as the diving response and provide an ideal
example of integrative physiology. The diving reflex can be stimu-
lated in the practical laboratory setting using breath holding and facial
immersion in water. Our undergraduate physiology students complete
a laboratory class in which they investigate the effects of stimulating
the diving reflex on cardiovascular variables, which are recorded and
calculated with a Finapres finger cuff. These variables include heart
rate, cardiac output, stroke volume, total peripheral resistance, and
arterial pressures (mean, diastolic, and systolic). Components of the
diving reflex are stimulated by 1) facial immersion in cold water
(15°C), 2) breathing with a snorkel in cold water (15°C), 3) facial
immersion in warm water (30°C), and 4) breath holding in air.
Statistical analysis of the data generated for each of these four
maneuvers allows the students to consider the factors that contribute
to the diving response, such as the temperature of the water and the
location of the sensory receptors that initiate the response. In addition
to providing specific details about the equipment, protocols, and
learning outcomes, this report describes how we assess this practical
exercise and summarizes some common student misunderstandings of
the essential physiological concepts underlying the diving response.
diving; bradycardia; heart rate; total peripheral resistance; reflex
control of arterial pressure
THIS REPORT DESCRIBES how undergraduate physiology students
can record, measure, and analyze changes in cardiovascular
variables during stimulation of the diving reflex in a 3-h
practical laboratory class. It describes the equipment and meth-
odology used, summarizes the physiological concepts that
could be taught before and after this practical class, and
provides learning objectives, a sample assessment question, a
class data set, and suggestions for using the practical as an
inquiry-based laboratory class.
Background
The diving response, also referred to as the diving reflex,
was initially described for the porpoise in 1941 (22). In
mammals, it is a sequence of physiological adjustments that
involve the nervous, cardiovascular, and respiratory systems
acting in a manner that promotes O2 conservation [through
decreased heart rate (HR) and cardiac output (CO)] and the
diversion of blood flow to vital organs such as the brain and
heart during underwater submersion (7, 17). In an undergrad-
Address for reprint requests and other correspondence: J. Choate, Dept. of
Physiology, School of Biomedical Sciences, Monash Univ., Clayton, VIC
3800, Australia (e-mail: Julia.Choate@monash.edu).
1043-4046/14 Copyright © 2014 The American Physiological Society
Downloaded from www.physiology.org/journal/advances (186.141.138.240) on October 21, 2019.
Sourcebook of Laboratory Activities in Physiology
uate physiology laboratory class, the diving reflex can be
stimulated experimentally by engaging students in breath hold-
ing exercises, in combination with facial immersion in water
(20). Variables such as water temperature and the area of facial
stimulation (e.g., intranasal, forehead, and/or cheek) can be
altered to investigate the different triggers of the diving reflex
and to observe specific cardiovascular adjustments (8, 35). The
physiology practical exercise reported in this article uses the
Finapres finger cuff to record cardiovascular variables (14, 21)
and is based on the “simulated human diving” teaching labo-
ratory exercise previously described by Hiebert and Burch
(20), in which only the effects on HR were examined. The
Finapres finger cuff, together with Beatscope software, allows
continuous and simultaneous recording and calculation of
multiple cardiovascular variables. These include HR, arterial
blood pressure (BP) [diastolic BP (DBP), systolic BP (SBP),
and mean arterial pressure (MAP)], CO, stroke volume (SV),
and total peripheral resistance (TPR; 14, 21). The unique
aspect of the diving reflex is that it involves the simultaneous
effects of both sympathetic and parasympathetic nerve activa-
tion on the cardiovascular system to alter HR, CO, arterial
pressure, and TPR (17, 28). This profile of responses contrasts,
for example, with the arterial baroreceptor reflex, which in-
volves reciprocal changes in sympathetic and parasympathetic
activity. By comparing these reflexes, students can learn that
the autonomic responses to physiological challenges can be
patterned in different ways.
Learning Objectives
After completing this practical laboratory class and writing
the associated scientific report, students should be able to:
1. Collect, analyze, and plot the cardiovascular data resulting
from the maneuvers in which components of the diving reflex
are stimulated.
2. Discuss the cardiovascular responses (for CO, HR, TPR,
MAP, DBP, SBP, and SV) observed when components of the
diving reflex are stimulated by facial immersion in cold water,
breathing with a snorkel in cold water, facial immersion in
warm water, or breath holding in air (apnea).
3. Describe the neuronal pathways that mediate the cardio-
vascular responses observed when the diving reflex is stimu-
lated and explain the physiological mechanisms underlying
these.
4. Communicate the background, results obtained (including
statistical analysis), and conclusions drawn from the practical
exercise in the form of a scientific paper.
Activity Level
This activity is suitable for undergraduate science, biomed-
ical science, or medical and allied health students, but it could
355
Sourcebook of Laboratory Activities in Physiology
356 STIMULATING THE DIVING REFLEX IN HUMANS
also be used in a simplified form (in which only HR and/or BP
are measured) for high school students of biology. We run this
practical class for a third-year physiology science class (Clin-
ical and Experimental Cardiovascular Physiology course) of
⬃300 students (in groups of ⬃40 students) of 19 –28 yr old.
Prerequisite Student Knowledge or Skills
Prequisite student knowledge. Before doing this activity,
students should have a basic understanding of the human
cardiovascular, respiratory, and autonomic systems. The spe-
cific physiological concepts required are listed below:
• CO is determined by the rate at which the heart contracts
(HR) and the volume of blood ejected with each cardiac
contraction (SV).
• MAP of the systemic circulation is altered by changes in CO
and TPR.
• The peripheral nervous system consists of cranial, spinal,
somatic motor, and autonomic nerves.
• Short-term control of cardiac function (SV and HR) and
TPR, and thus MAP, is in part achieved by the central
nervous system processing incoming (afferent) neuronal
information from a range of sensory receptors. These sen-
sory receptors include arterial baroreceptors, chemorecep-
tors, and atrial and pulmonary volume receptors. Changes in
the activity of afferent nerves alters the outgoing (efferent,
autonomic) neuronal activation from the cardiovascular cen-
ters in the medulla oblongata and thus the activity of post-
ganglionic autonomic nerves that innervate effector organs
(the heart) and tissues (blood vessels).
• Increased activation of parasympathetic nerves (in the va-
gus) and decreased activation of sympathetic nerves inner-
vating the sinoatrial node slows HR by decreasing the rate
of the spontaneous generation of cardiac pacemaker action
potentials.
• Differential activation of sympathetic nerves innervating
arterioles leads to regional enhancement of the contraction
of vascular smooth muscle (i.e., arteriolar constriction).
• Breathing requires the activation of the diaphragm, intercostal,
and accessory thoracic skeletal muscles via somatic motor
neurons projecting from the respiratory center in the medulla
oblongata of the brain stem.
• Apnea is the cessation of breathing, which can be voluntarily or
reflexly induced.
• Arterial chemoreceptors respond to changes in PO2 (PaO2) and
PCO2 (PaCO2). Central chemoreceptors are activated by acidifi-
cation of blood (pH), which occurs when PaCO2 increases. Apart
from effects on respiratory drive, activation of arterial and
central chemoreceptors also alters cardiovascular function
through activation of the autonomic nervous system.
Prequisite student skills. Students should know how to do
the following:
• Use the recording software (start and stop recording, add
comments to the recording trace, save the file, and extract
and enter the data into an Excel spreadsheet).
• Calculate the mean and SD and plot data using the appro-
priate software [e.g., Excel and Graphpad Prism (18)].
• Calculate (for each variable) the percent change from the
baseline values in response to each test condition.
Advances in Physiology Education • doi:10.1152/advan.00125.2013 • http://advan.physiology.org
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• Perform statistical analysis of these data. [For instructors:
this component could be simplified if the statistical analysis
was performed and provided to the students, but this is
something that students can easily complete if they are
provided with the appropriate instructions].
• Interpret the results of the statistical analysis of class data
(i.e., what does P ⫽ 0.01 mean?).
• Source and cite research articles.
Time Required
The entire experimental protocol, which is composed of four
maneuvers in which components of the diving reflex are
stimulated, can be completed during a 3-h practical class.
METHODS
Each of our practical classes have ⬃40 students, and the practical
is run 8 times (we had 317 students in 2013). Students work in 10
groups of 4 students/group, where each group member is assigned one
of the following team roles: 1) a voluntary experimental subject who
completes the four experimental maneuvers; 2) a computer operator
who obtains the recordings of the data; 3) a data manager who extracts
and calculates the values of the specific cardiovascular variables from
the record of the data; or 4) a team manager who manages the
experimental protocol, including acting as a time keeper, and coordi-
nates the other group members.
Equipment and Supplies
The photograph in Fig. 1 shows how some of the equipment for this
practical class is organized on the laboratory bench. Each student
group is provided with the following equipment and supplies:
• A Finapres finger cuff recording system, Finometer MIDI, with
Beatscope software (14, 21)
• A 10-liter plastic basin containing ⬃7.5 liters of tap water at room
temperature
• A thermometer
• A timer (in min:s)
• A plastic nose clip
• A snorkel (no mask) or a plastic breathing tube (vinyl, internal
diameter: 10 mm) (a snorkel is preferable to a plastic breathing
Fig. 1. An experimental subject immersing his face in 7.5 liters of water
(15°C) placed in a 10-liter plastic basin. The subject breathes through a plastic
tube (held out of the water with their left hand). The Finapres finger cuff is
placed on the index finger of the right hand. A time keeper taps the subject on
the shoulder every 10 s and states the time out loud. A cloth towel is positioned
nearby for drying the face once the experimental maneuver is complete.

STIMULATING THE DIVING REFLEX IN HUMANS
tube. If a plastic breathing tube is used, it must be flexed or bent to
prevent the end of the tube going under the water)
• Paper or cloth towels
• Ice and warm tap water
• Excel and Graphpad Prism software on the desktop computer for
compiling and plotting the data and performing tests of signif-
icance (18) [free software called “R” can also be used for these
processes (31)]
What can you do if a Finapres recording system is not available?
There are other systems available for recording CO. PhysioFlow and
BIOPAC Systems NICO100C (with AcqKnowledge software) hemo-
dynamic monitors use impedance cardiography to provide noninva-
sive and continuous measurements of CO (29). It should be noted that
most of these systems (including the Finapres recording system) are
expensive.
HR can be calculated using palpation or with a different HR
recording system, such as a Polar monitor (30). Inexpensive digital
HR monitors are available from many sports stores.
Arterial BP and HR can be noninvasively and continuously re-
corded with a BIOPAC systems NIBP100D finger cuff (6, 16).
Inexpensive digital BP monitors (e.g., Omron) can be purchased from
a pharmacy (26). These provide measurements of arterial BP (MAP,
SBP, and DBP) and pulse rate. AD Instruments has a system that
records SBP and HR (1). A manual sphygmomanometer could also be
used. When used for the experimental protocol we describe here,
students should be made aware that these digital and manual BP
monitors (which do not record BP continuously) can take ⬃30 s to
generate values. This becomes problematic if you are recording a
response that is relatively transient.
Use of Voluntary Human Subjects for the Diving Reflex Protocol
The use of human subjects for these practical laboratory experi-
ments may require ethical approval from your institution’s human
research committee. Adopters of this activity are responsible for
obtaining permission for human or animal research from their home
institution. For a summary of Guiding Principles for Research Involv-
ing Animals and Human Beings, please see www.the-aps.org/mm/
Publications/Ethical-Policies/Animal-and-Human-Research. At Mo-
nash University, Human Research Ethics approval is not required for
undergraduate teaching involving the participation of students. How-
ever, students are told that their participation as experimental subjects
in the diving reflex protocol is voluntary and that they can withdraw
at any time.
Instructions
Step 1. Allocate the team roles for each member of your group (i.e.,
voluntary experimental subject, computer operator, data manager, and
team manager). (For instructors: in our classes, we do not specify the
sex of the subject, principally so that each group of 4 students can
select the person most capable of performing the protocol. However,
Advances in Physiology Education • doi:10.1152/advan.00125.2013 • http://advan.physiology.org
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Sourcebook of Laboratory Activities in Physiology
357
this could be taken into consideration, and we do discuss this as a
potential limitation of the study.)
Step 2. To dissect out the various components of the diving reflex,
you will investigate the effects of the following sequential experimen-
tal maneuvers on cardiovascular variables: 1) facial immersion in cold
water (15°C; manuever 1), 2) breathing with a snorkel in cold water
(15°C; manuever 2), 3) facial immersion in warm water (30°C;
manuever 3), and 4) breath holding in air (manuever 4). The same
subject is used for all of the experimental maneuvers (see Fig. 2). The
subsequent steps specify how to complete the experimental protocol.
Step 3. The experimental subject remains sitting, with elbows
resting on the laboratory bench, for all of the experimental maneuvers.
They should not alter their position (i.e., stand) during the protocol as
this will lead to altered baroreceptor function. Encourage them to be
relaxed and situate them so that they are unable to see the computer
trace of the cardiovascular recordings.
Step 4. Place ⬃7.5 liters of tap water into the plastic basin. Adjust the
temperature of this water to 15°C by adding warm tap water or ice to the
water. Please note that the room temperature during the experimental
protocol is maintained at 21 ⫾ 1°C.
Step 5. Fit the Finapres finger cuff onto the index finger of the
dominant hand of the experimental subject. Calibrate the finger
cuff and fix the height indicator into place on the subject’s chest at
the level of the heart. Enter the demographic information about the
experimental subject into the computer (name, age, weight, and
sex), using Beatscope software. This software is set up to capture
recordings and calculations of the following cardiovascular vari-
ables: HR, CO, MAP, DBP, and SBP. [For instructors: in our
practical class, students calculate SV and TPR (the Finapres
system can do this, but performing these calculations will help
students understand the relationships among MAP, CO, HR, SV,
and TPR). A description of the Finapres technology can be found
in a previous teaching publication (21)]. Assign an appropriate
name to this file and continuously record cardiovascular variables
for the experimental protocols. You will be adding comments to
this recording as you complete the protocols.
Step 6. With the subject in the relaxed position (elbows on bench
with the basin in front of them), record cardiovascular variables for at
least 5 min. Label this as “test run” on the computer recording. The
experiment protocol in which components of the diving reflex are
stimulated can now be performed (see step 2 and Fig. 2). Before
commencing the four maneuvers, complete a test run for facial
immersion in water so that the subject can become familiar with the
experience. This is an important step as it will reduce the effects of
stress on the cardiovascular recordings when the actual experiment is
carried out. The subject puts their face in the water for ⬃5 s, with the
water covering the forehead, eyes, and nose (see Fig. 1). Before facial
immersion in water, the subject should take a moderate inspiration,
making sure that they do not perform a Valsalva maneuver [i.e.,
exhaling against closed airways, leading to increased thoracic pressure
and reduced venous return (37)]. After this trial facial immersion, the
Fig. 2. Protocol schema outlining the se-
quence events followed during the four ma-
neuvers that are used to stimulate compo-
nents of the diving reflex in the same subject
during the practical class.
Sourcebook of Laboratory Activities in Physiology
358 STIMULATING THE DIVING REFLEX IN HUMANS
subject rests for 5 min to allow values to return to a steady baseline
before proceeding with maneuver 1.
Step 7. To commence maneuver 1, record cardiovascular variables
for at least 5 min to obtain baseline values. The experimental subject
then places their face into the cold water (15°C) for 30 s. Label the
computer recording as “stimulated diving reflex maneuver 1 - cold
water.” This represents the control maneuver for the experiment.
Step 8. A time keeper taps the subject on the shoulder every 10 s
and states the time out loud, allowing the subject to keep track of the
time and to remain calm (see Fig. 1).
Step 9. The subject removes their face from the water. Use a cloth
or paper towel to dry the subject’s face. Label the computer recording
as “recovery” and continue recording for 5 min or until the cardio-
vascular variables stabilize.
Step 10. Repeat steps 7–9 for maneuver 2 (breathing with a snorkel
in cold water), manuever 3 (facial immersion in warm water), and
manuever 4 (breath holding in air). For maneuver 3, the water
temperature in the basin should be increased to 30°C by the addition
of warm water; for maneuver 4, the subject holds their breath for 30
s (with their head out of water but still leaning over the basin). Again,
for maneuver 4, a Valsalva maneuver should not be performed. The
subject rests for at least 5 min between each of the maneuvers in
which they stimulate components of the diving reflex (see Fig. 2).
Step 11. Once the four experimental maneuvers are completed, stop
the recording and save the file. The data manager in the team can now
extract values for each of the cardiovascular variables (HR, CO, MAP,
DBP, and SBP) at 1, 3, and 5 min during the relevant baseline period
and at 0, 15 and 30 s after commencing each of the four maneuvers in
the protocol. The experimental team should work together to calculate
SV and TPR and enter all of the cardiovascular variables into a class
spreadsheet (using Excel) in preparation for plotting the data and
performing statistical analyses. A sample data collection sheet for
manuever 1 is shown in Table 1. [For instructors: the class data set
should be scrutinized at this stage to look for data calculation or entry
errors. In addition, occasionally subjects show a marked stress re-
sponse (increased HR) for each condition except the breath holding in
air. It is better to remove these subjects from the subsequent analysis
and to explain the rationale for doing this to the students. A simpli-
fication that could be used is to analyze only MAP and not SBP and
DBP].
Step 12. Using the class data, average the first two time points
during the baseline period (1 and 3 min) and the time points 15 and
30 s after commencing the experimental maneuver for each exper-
imental subject and for the four maneuvers in the protocol. We do
not use the values at the 5-min mark of the baseline period or the
values immediately after commencing the experimental maneuver
(0 s) because we find that most students display an anticipatory
response, which can dilute the effect. The rationale for this is
explained to the students. Determine the mean and SD for each
cardiovascular variable for the class data [For instructors: it is best
to use SD because it provides a measure of dispersion that, unlike
Table 1. A sample data collection sheet showing the formatting and data for a single experimental subject that has
undergone facial immersion in cold water (i.e., maneuver 1)
SBP, mmHg DBP, mmHg MAP, mmHg
Baseline
1 min 129 69 88
3 min 132 71 88
5 min 132 73 89
Maneuver 1
0s 146 79 99
15 s 168 91 110
30 s 168 92 114
SBP, systolic blood pressure; DBP, diastolic blood pressure; MAP, mean arterial pressure; HR, heart rate; SV, stroke volume; CO, cardiac output; TPR, total
peripheral resistance.
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SE, is not dependent of the number of observations on which it is
based.]
Step 13. We use Graphpad Prism for statistical analysis. There are
two steps to the statistical analysis (steps a and b).
STEP A. Perform repeated-measures ANOVA with factors of treat-
ment (the four experimental maneuvers) and time (before and during
the stimulations of the diving reflex) and their interaction followed by
Student’s paired t-tests with a Bonferroni correction to account for
multiple comparisons. [For instructors: ask your students to explain
the characteristics of this data set that make it appropriate for use of
repeated-measures ANOVA (i.e., all four maneuvers were performed
on the same experimental subject) and the critical differences between
repeated-measures ANOVA and conventional ANOVA (i.e., the po-
tential inclusion of both between-subjects and within-subjects factors
and the potential for use of corrections to protect against the increased
risk of type 1 error arising from compound asymmetry). Convention-
ally, post hoc tests are only performed if the results of preliminary
ANOVA shows some level of statistical significance (conventionally,
P ⱕ 0.05). Students should be made aware of these scientific con-
ventions, the reasoning that underlies them, and what a P value
actually represents].
Post hoc analysis allows comparison of the baseline values for each
protocol, to determine if resting values have changed across time
(acclimatization). [For instructors: this reinforces the importance of
well-controlled experiments and introduces the concept of acclimati-
zation to experimental conditions across a study. This information
may help the students to explain why expected reductions in the
magnitude of the responses might not be observed].
Post hoc analysis also determines which variables significantly
changed from baseline in response to each protocol, to establish if the
maneuvers used to stimulate the diving reflex had effects (i.e., facial
immersion in cold water, breathing with a snorkel in cold water, facial
immersion in warm water, and breath holding in air).
STEP B. Perform repeated-measures one-way ANOVA, using the
data expressed as percent changes from baseline, to determine how
altering the test conditions (i.e., maneuvers 2– 4) changes the response
compared with facial immersion in cold water (i.e., maneuver 1, the
control). [For instructors: encourage students to consider the variabil-
ity in the data and why expressing data as percent changes reduces this
variability].
If the results of ANOVA indicate significant differences be-
tween mean values, use Dunnett’s post hoc test to determine
whether the magnitudes of the effects of each of the partial stimuli
(maneuvers 2– 4) differed from those to facial immersion in cold
water (maneuver 1, the control). [For instructors: you may choose
to advise students to reject the null hypothesis, i.e., that the
magnitudes of the responses to each of the partial stimuli (maneu-
vers 2– 4) did not differ from that to facial immersion in cold water
(maneuver 1) if P values from the Dunnett’s test are ⱕ0.05 (or
some other critical value you might choose). Alternatively, you
HR, beats/min SV, ml CO, l/min TPR mmHg·l⫺1·min⫺1
80 84 6.6 14
78 85 6.5 14
86 83 7.0 14
95 80 7.5 14
66 81 5.3 22
68 84 5.7 23
 Sourcebook of Laboratory Activities in Physiology
STIMULATING THE DIVING REFLEX IN HUMANS 359

10 between the responses to maneuvers 2– 4 compared with those to

maneuver 1, data from 8 –10 experimental subjects are required. For
example, with a sample size of eight and a 10% SD (␣ ⫽ 0.05), a 15%
Recovery mean difference in HR can be detected with 80% power. It is also
useful to generate a larger data set over time, to demonstrate the
expected responses more clearly. Data should be excluded from the
analysis if the subject’s HR and CO progressively increased during
the stimulated diving reflex and did not return to baseline during the
Cardiac recovery phase. These responses are presumed to result from the
Output effects of stress and therefore do not reflect the diving reflex. In a class
l/min with 10 experimental subjects, we find that the data from 1 or 2
subjects can be removed without seriously affecting the statistical
Baseline outcomes. Over the years, during the running of this class, we have
removed an average of 1.3 (SD 0.8) subjects (n ⫽ 16 classes, range:
0 –2 subjects) from a data set due to these stress effects.
Safety Considerations
Facial Immersion in cold water
0 Appropriate Occupational Health and Safety procedures are fol-
250 lowed in the laboratory. Students who have cardiovascular disease, a
previous history of fainting, are pregnant, have any kind of infection,
or are taking any medication that could alter cardiovascular responses
should be excluded as experimental subjects.
Care must be taken to prevent the spread of infection. Between
Arterial practical classes (and thus experimental subjects), the snorkel and
Pressure plastic basin are cleaned with Sonidet (a bacteriostatic cleaning fluid
SBP designed for the medical field) and the liquid antiseptic Dettol. In
mmHg
and addition, the benches are wiped down with 70% (vol/vol) ethanol and
Dettol, and the inner lining of the Finapres finger cuff is cleaned with
Heart
MAP
alcohol swabs.
Rate
bpm DBP
RESULTS
HR
Effects of Stimulating the Diving Reflex on Cardiovascular
Variables

0 The raw data trace shown in Fig. 3 shows that facial

immersion in cold water (maneuver 1) causes HR and CO to
decrease and arterial pressures (MAP, DBP, and SBP) to
Time (hours:minutes:seconds)
increase. These changes do not develop immediately but be-
Fig. 3. A Finapres recording showing the effects of facial immersion in cold come more pronounced across the 30 s of the maneuver. Also
water (15°C) on cardiac output (CO), heart rate [HR; in beats/min (bpm)] and shown in this experimental recording is an initial increase in
arterial pressure [mean arterial pressure (MAP), diastolic blood pressure
(DBP), and systolic blood pressure (SBP)]. HR (and thus CO), which is presumed to be an anticipatory
response involving “central command activation” (11). Finally,
during the recovery phase, the cardiovascular variables return
could use this as an opportunity to discuss the meaning of P and the to baseline fairly rapidly.
advantages of considering its absolute value in evaluating the
strength of evidence against the null hypothesis (9, 10)].
Step 14. Use the statistical analysis of the class data as the basis for Table 2. Effects of the stimulation of the diving reflex on SV,
a scientific report. SBP, and DBP
Troubleshooting Percent Percent Change Percent Change
Stimulated Diving Protocol Change in SV in SBP in DBP
Some students who have limited swimming or snorkeling experi-
ence may find facial immersion in water a difficult task to accomplish. Facial immersion in cold
Furthermore, unfamiliarity with the maneuvers can contribute to water (maneuver 1) ⫺6 (19) 27 (14)††† 40 (23)†††
Breathing with a snorkel
excessive anticipatory anxiety (i.e., elevated HR). At the beginning of in cold water
the class, a teacher or practical class technician should demonstrate (maneuver 2) 1 (13) 7 (7)** 10 (11)**
the four different maneuvers for stimulating the diving reflex to the Facial immersion in
class. Also, as described above, the subject should complete a test run warm water
of the first maneuver; otherwise, the difference between maneuvers 1 (maneuver 3) 14 (26) 10 (7)*†† 15 (13)*†
and 2 could be due to a training effect. Breath holding in air
We always discuss the concepts of sample size and variability with (maneuver 4) 4 (16) 4 (10)** 7 (16)*
the students. The class data presented in this report represent 10 Data are expressed as means (SD) of percent changes from baseline values
experimental subjects from a single practical class (see Table 2 and for each maneuver. *P ⱕ 0.05 and **P ⬍ 0.01 for maneuvers 2– 4 vs.
Fig. 4). To generate a data set with sufficient power to detect manuever 1 (control group); †P ⱕ 0.05, ††P ⬍ 0.01, and †††P ⬍ 0.001
responses to the various experimental stimuli as well as differences compared with baseline values.

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Sourcebook of Laboratory Activities in Physiology
360 STIMULATING THE DIVING REFLEX IN HUMANS

CW T WW A CW T WW A
0 0

% change in CO
% change in HR
-10 -10

-20 † -20
**
††† *** ††
-30 ††† -30
*
Fig. 4. Effects of facial immersion in cold water (CW; * *
15°C), breathing with a snorkel in cold water (T), facial -40 -40
immersion in warm water (WW; 30°C), and breath
††† †††
holding in air (A) on HR, CO, MAP, and total peripheral -50 -50
resistance (TPR). Data are expressed as mean percent
changes from the corresponding values during the base-
line period with SD (n ⫽ 10 students). †P ⱕ 0.05, ††P ⬍
0.01, and †††P ⬍ 0.001 compared with baseline values; 60 ††† 200
*P ⱕ 0.05, **P ⬍ 0.01, and ***P ⬍ 0.001 for maneu-
†††

% change in TPR
% change in MAP
vers 2– 4 vs. maneuver 1 (facial immersion in cold water, 150
control).
40
* 100 **
** ††
* ††
†
20 ** †††
50 **

0 0
CW T WW A CW T WW A

The averaged experimental data from 10 subjects from a
single practical class are shown in Table 2 and Fig. 4. Facial
immersion in cold water (maneuver 1) reduced HR and CO
and increased TPR, MAP, SBP, and DBP (P ⱕ 0.05 for all
variables). Significant changes in cardiovascular variables
were observed in response to the other three maneuvers, in
which components of the diving reflex were stimulated.
Breathing with a snorkel in cold water (maneuver 2) reduced
HR and CO and increased TPR, MAP, SBP, and DBP (all
P ⱕ 0.05). Facial immersion in warm water (maneuver 3)
reduced HR and increased TPR, MAP, SBP, and DBP (all
P ⱕ 0.05), but the fall in CO did not reach statistical
significance (P ⫽ 0.2). Breath holding in air (maneuver 4)
caused a small, but statistically significant, fall in HR (P ⱕ
0.05) but did not affect any other variable. SV was not
significantly affected by any of the maneuvers (Table 2).
Dunnett’s post hoc test was then performed to compare the
effects of maneuvers 2– 4 (i.e., the partial stimuli) with those of
maneuver 1 (the control). The results provide evidence that the
responses to maneuvers 2– 4 were blunted compared with those to
maneuver 1 (facial immersion in cold water). For maneuvers 2– 4,
HR, TPR, SBP, DBP, and MAP responses were all less than those
to manuever 1 (all P ⱕ 0.05). The fall in CO was attenuated in
response to facial immersion in warm water and breath holding in
air (both P ⱕ 0.05). However, the fall in CO was not significantly
less in response to breathing with a snorkel in cold water com-
pared with facial immersion in cold water (P ⫽ 0.08). The change
in SV was similar for all maneuvers (Table 2).
Interpretation of Class Data
The statistical analysis of our stimulated diving reflex ex-
periment supports the proposition that facial immersion in cold
water stimulates neuronal pathways involved in the diving
reflex. The other maneuvers (i.e., breathing with a snorkel in
cold water, facial immersion in warm water, and breath holding
in air) were used to test the effects of activation of subsets of
these pathways (Fig. 5).
Evidence from human and animal experimental studies in-
dicates that the diving response is mediated by a network of
sensory, central, and autonomic neurons that control the respi-
ratory and cardiovascular systems (7, 17, 20, 27). This makes
practical classes, in which the diving reflex is stimulated, ideal for
teaching integrative physiology to students of undergraduate

Fig. 5. A–D: schematic illustrating the sensory inputs and neuronal pathways involved in the cardiovascular and respiratory responses during the four maneuvers
that stimulate components of the diving reflex. Solid lines represent neuronal excitation, and dashed lines represent neuronal inhibition. The thickness of the lines
is indicative of the relative strength of the activation (i.e., thicker lines represent stronger activation). RVLM, rostral ventrolateral medulla; IML, intermediolateral
column; PaO2, arterial PO2; PaCO2, arterial PCO2. A: during facial immersion in cold water (maneuver 1, shown in gray), due to activation of temperature and
touch-sensitive receptors on the face, apnea, and chemoreceptors, large falls in HR and CO are induced via parasympathetic activation, and increases in TPR
due to sympathetic activation are observed. B: in response to breathing through a snorkel in cold water (maneuver 2, shown in green), in which the contribution
of apnea and subsequent activation of chemoreflexes is removed, the integrated efferent output is reduced, resulting in an attenuation of the cardiovascular
responses that make up the dive response. C: facial immersion in warm water (maneuver 3, shown in red), in which input from facial temperature-sensitive
receptors is significantly reduced, also attenuates the signal driving the dive response, demonstrating the importance of water temperature in the initiation of the
dive response. D: finally, if the only stimulus is apnea and the resultant activation of chemoreflexes, illustrated by breath holding in room air (maneuver 4, shown
in blue), the integrated efferent output and cardiovascular responses are modest compared with those to facial immersion in cold water. Thus, on its own, apnea
does not elict a powerful dive response, but it does reinforce the impact of facial immersion in cold water, given the attenuated response to breathing through
a snorkel in cold water.

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 Sourcebook of Laboratory Activities in Physiology
STIMULATING THE DIVING REFLEX IN HUMANS 361

A B

C D

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Sourcebook of Laboratory Activities in Physiology
362 STIMULATING THE DIVING REFLEX IN HUMANS
science (physiology) because it encourages them to consider all
of the pathways involved:
• The various sensory inputs that detect and trigger the re-
sponse
• The afferent pathways conveying information to the central
nervous system
• Integration by the central nervous system
• The efferent pathways to the effector organs
• The corresponding responses of the effector organs
Cardiovascular responses to stimulation of the diving reflex.
The cardiovascular responses induced by activation of the
diving reflex include reductions in HR and CO and increases in
TPR and MAP (2, 7, 17, 24, 28). All of these cardiovascular
effects have been illustrated during facial immersion in cold
water in our practical class and in experiments involving
human divers (28).
Since there is little evidence of a change in SV during real
diving or when the diving reflex is stimulated, the reduction in
CO must result from the associated bradycardia (17, 28, 32).
Although not observed in the class data set presented, in the
initial classes that were run, we often observed a small, but
significant, decrease in SV with breath holding in air, which
would contribute to the reduction in CO. This small reduction
in SV during breath holding was likely due to a Valsalva
maneuver by the experimental subjects. That is, this maneuver
increases intrathoracic and right atrial pressures, leading to
decreased venous return and decreased SV, via the Frank-
Starling mechanism (37). We now provide students with spe-
cific instructions at the beginning of the class about how to
avoid performing a Valsalva maneuver. As shown in the
presented class data, this has resulted in the SV remaining
unaffected during stimulation of the diving reflex.
Students should come to the practical class with the knowl-
edge that the changes in the cardiovascular variables recorded
during each experimental maneuver result from the effects of
sympathetic nerve activation on the arterioles and from
changes in both sympathetic and parasympathetic nerve acti-
vation on the heart. To write their scientific report, they will
need to investigate the neuronal pathways that underlie these
autonomic effector responses.
Neuronal pathways that underlie the cardiovascular re-
sponses to stimulation of the diving reflex. Facial temperature
and touch-sensitive receptors, chemoreceptors, baroreceptors,
pulmonary stretch receptors, and atrial receptors project via
afferent nerves to the nucleus tractus solitarius (NTS) in the
medulla oblongata (17, 27). Activity in these afferent path-
ways, together with those arising from central chemoreceptors,
can be altered when the diving reflex is stimulated or during
real diving. As shown in the schematics in Fig. 5, from the
NTS, neurons project within the medulla oblongata to the
cardiac parasympathetic center (nucleus ambiguus), the vaso-
motor center [rostral ventrolateral medulla (RVLM)], and the
respiratory center (17, 27). For sympathetic control of the
cardiovascular system during the diving reflex, neurons in the NTS
are activated. These neurons project via interneurons to the
excitatory (glutamatergic) neurons in the RVLM. The RVLM
controls sympathetic nerve activation via bulbospinal neurons
that project to sympathetic preganglionic neurons via the in-
termediolateral column of the spinal cord (17, 27). For para-
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sympathetic control of the cardiovascular system during the
diving reflex, neurons in the NTS are activated, leading to the
activation of preganglionic parasympathetic neurons that proj-
ect directly from the nucleus ambiguus (in the medulla oblon-
gata) and then synapse with postganglionic parasympathetic
neurons within intrinsic cardiac ganglia. These postganglionic
neurons mainly innervate the sinoatrial node and atrioventric-
ular node, and their activation decreases HR and slows the rate
of atrioventricular impulse conduction (17, 20, 25, 32).
Apnea can be induced voluntarily, in which case signals are
generated by the cerebral cortex and sent via the hypothalamus,
or reflexly (involuntary), via activation of temperature-sensi-
tive receptors on the face (17, 27). In either case, the signals,
induced by apnea, are relayed to the dorsal respiratory group in
the NTS, leading to excitation of the RVLM and nucleus
ambiguus and increasing autonomic activation of the heart and
blood vessels (see Fig. 5) (3, 17). The central neuronal path-
ways involved in chemoreceptor-mediated respiratory re-
sponses are not well understood (27). However, peripheral
chemoreceptors are more sensitive to changes in PaO2 and
central chemoreceptors respond to the acidification of blood
due to increased PaCO2 (17, 27).
During diving, a decrease in the activity of pulmonary
stretch receptors could contribute to the development of the
diving bradycardia (4, 5). The pulmonary stretch receptors
would be important only if lung volume is changing (34). In
this practical class, there is usually no change in lung
volume during the stimulated diving protocols, with apnea
held at a moderately increased lung volume. However, it is
possible that some experimental subjects may breathe out
into the water. Lung volume can influence the absolute
changes in HR and BP during facial immersion in water.
However, differences in lung volume (60% compared with
85% of vital capacity) have little impact on the pattern or
timing of the cardiovascular responses (3).
Altering the sensory inputs. During facial immersion in cold
water (maneuver 1 for our laboratory class), the dive reflex is
initiated by facial temperature and touch-sensitive receptors
and apnea. Afferent impulses then project via cranial nerves
(mainly the trigeminal) to the NTS, leading to 1) inhibition of
the medullary respiratory center, 2) activation of the RVLM
in the vasomotor center, and 3) activation of the nucleus
ambiguus in the cardiac parasympathetic center (17, 27). All
three of these neuronal pathways will be operating at the same
time (see Fig. 5A) (27). In addition, baroreceptor input will be
temporarily overridden.
First, inhibition of the medullary respiratory center overrides
the autonomic centers in the medullary rhythmicity area (the
ventral and dorsal respiratory groups), causing the cessation
of breathing and inhibition of the diaphragm, intercostal and
accessory thoracic muscles, thus preventing the aspiration of
water (15, 17).
Second, excitation of the RVLM causes increased activation
of sympathetic nerves innervating blood vessels, elevating
TPR and MAP (23). This activation is regionally selective,
with increased sympathetic activation to nonessential organs,
including the gut and skin. Increased muscle sympathetic nerve
activity induces vasoconstriction of muscle blood vessels (12).
It has been shown experimentally in human subjects that facial
immersion in cold water increases muscle sympathetic nerve
activity, with apnea alone producing much smaller increases (12).

STIMULATING THE DIVING REFLEX IN HUMANS
These experiments demonstrate the importance of the facial re-
ceptors and trigeminal afferents in sympathetically mediated car-
diovascular responses during stimulation of the diving reflex.
Third, the simultaneous excitation of the nucleus ambiguus
leads to the activation of cardiac parasympathetic nerves and
the reduction in HR, often referred to as “diving bradycardia”
(17, 19, 32).
Breathing with a snorkel in cold water (maneuver 2 for our
laboratory class; see Fig. 5B) elicits activation of the facial
receptors, without apnea. We found that this stimulus caused
smaller changes in cardiovascular variables (HR, MAP, DBP,
SBP, and TPR, with a trend for a reduction in the fall in CO)
than facial immersion in cold water with breath holding (ma-
neuver 1). This supports the idea that the apnea and/or its
consequences (hypoxia and hypercapnia) contribute(s) to the
cardiovascular responses to facial immersion in cold water but
that an attenuated response can be driven by facial temperature
and touch-sensitive receptors alone.
We found that facial immersion in warm water (maneuver 3)
resulted in smaller changes in cardiovascular variables (HR,
CO, MAP, DBP, SBP, and TPR) than facial immersion in cold
water (maneuver 1). This suggests that water temperature is an
important stimulus for driving the dive response (Fig. 5C) and
is consistent with experimental evidence showing that there is
greater activation of facial temperature-sensitive receptors
when they are exposed to water at lower (10 –15°C) than
higher temperatures (17).
To extract the relative contributions of facial immersion in
water and breath holding to the cardiovascular responses dur-
ing the diving reflex, we compared the responses to breath
holding during facial immersion in cold water (maneuver 1)
versus breath holding in air (maneuver 4, i.e., without facial
immersion). Our results show that breath holding in air only
caused a minor fall in HR (⬃7 beats/min) compared with facial
immersion in cold water (⬃27 beats/min), with no other
detectable cardiovascular response (Fig. 4). This observation
demonstrates that apnea alone is not a primary driving force for
the dive reflex (Fig. 5D) but that apnea does reinforce the
response to facial immersion in cold water (Fig. 5A).
In conclusion, these experiments demonstrate that humans,
like aquatic animals, exhibit a dive response. It was established
that facial immersion in cold water and apnea, together, pro-
duced the greatest dive response. Thus, exposure of the face to
cold is a major stimulus. Furthermore, apnea on its own does
not drive the dive response but is a secondary reinforcing
stimulus, at least under the experimental conditions of facial
immersion in cold water.
Evaluation of Student Work
Each student group prepares a written practical report in the
format of a condensed scientific paper consisting of an Intro-
duction (maximum: 200 words), Results (no limit), and a
Discussion (maximum: 1,000 words). [For instructors: if you
have a smaller class, it is feasible for students to submit
individual reports.] They are provided with detailed instruc-
tions on the writing of a scientific report. The instructions
cover both the approach they should take and the format of the
scientific report. These have been developed over many years
by various members of staff of the Faculties of Medicine and
Science of Monash University. For the Introduction, students
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Sourcebook of Laboratory Activities in Physiology
363
are directed to start broadly and then to finish with specific
statements of the experimental aims and hypotheses. They are
provided with detailed instructions on the presentation of their
scientific results. For the Discussion, they are advised to start
with their major findings, and the conclusions drawn from
these, in the context of their aims and hypotheses. They should
follow this with arguments that link the findings and conclu-
sions. Specifically for the Discussion, the students are given the
following subheadings to incorporate:
1. Main findings
2. Population characteristics (are the baseline values within
the normal range for this population?)
3. Facial immersion in cold water (To describe the response
and discuss, based on the literature, the afferent and efferent
pathways involved and how these contribute to survival)
4. Each of the other test conditions (breathing with a snorkel
in cold water, facial immersion in warm water, and breath
holding in air) should be discussed in turn to, by subtraction,
identify the different components of the reflex and their im-
portance to the response to facial immersion in cold water
5. Limitations (a paragraph discussing the limitations of the
study and how they might impact on the results)
6. Conclusion (a paragraph about the physiological signifi-
cance of the experiment)
Students are provided with two starter references for their
report: Hiebert and Burch (20) and Foster and Sheel (17).
These instructions are guides to ensure that all major points are
covered. Students are allowed latitude in the order in which the
information is presented.
An essay question on the diving response is included on the
end-of-semester examination. This is not compulsory (i.e.,
students are given a range of essay topics to choose from). In
the past 3 yr, 10% (2012), 35% (2013), and 55% (2014) of the
students answered the diving response question, with average
marks of 75% (SD 21, 2012), 73% (SD 21, 2013), and 81%
(SD 5, 2014). An example of a diving response essay question,
for which 40 min is allowed for completion, is given below:
Describe the cardiovascular reflex responses to cold water
face immersion. Include the critical physiological variables
involved in the response:
1. The stimuli that elicit the response
2. The receptors and afferent and efferent pathways involved
3. The changes that occur in cardiovascular variables
If the conditions were altered as the subject immersed their
face in warm water, while breathing through a snorkel, how do
you think the responses would be altered? Why?
Misconceptions
Analysis of the students’ scientific reports and exam essays
revealed some common misunderstandings of the diving re-
sponse. These are summarized below, together with selected
student quotes. Many of these misunderstandings reflect lan-
guage skills rather than knowledge deficiencies. Furthermore,
some students use jargon (especially under exam conditions)
rather than scientific language.
Misconception 1. Misunderstanding of the anatomy and ac-
tions of the autonomic nervous system during the diving response:
Stimulation of chemoreceptors activates the sympathetic and
parasympathetic nervous systems, which causes blood vessels
to constrict.
Sourcebook of Laboratory Activities in Physiology
364 STIMULATING THE DIVING REFLEX IN HUMANS
This student did not understand that only sympathetic nerve
activation alters arteriolar diameter.
Trigeminal and mechanoreceptor stimulation increases sym-
pathetic and parasympathetic input to the vagus nerve.
Only parasympathetic nerves are within the vagus.
Misconception 2. The involvement, locations, and functions
of multiple types of sensory receptors (i.e., arterial chemore-
ceptors, facial temperature, and touch-sensitive receptors) dur-
ing the diving simulations:
The touch receptors on the forehead will elicit a cardiovas-
cular responseѧvia cold water acting on the carotid arch
This student has confused receptor location and type; the temper-
ature-sensitive receptors are on the face and the baroreceptors are
in the aortic arch and carotid sinus.
Skin receptors on the face relay messages to the brainѧ
What are skin receptors?
These nerves detect the decrease in temperature and trigger
the chemoreceptors to send information to the respiratory
centerѧ
This student had changes in temperature activating chemore-
ceptors.
Immersing your face in water causes the chemoreceptors on
the face to become stimulated.
Arterial chemoreceptors are activated during apnea because of
the evolving hypoxemia. This student might think that there are
facial chemoreceptors or that facial chemoreceptors sense tem-
perature.
Misconception 3. Some students did not seem to understand
the locations and functions of the arterial baroreceptors. Baro-
receptors respond to changes in arterial stretch/pressure by
altering their rate of firing, not by changes in receptor number:
The wetness of the cold water and pain causes the barore-
ceptors to respondѧ
ѧarterial receptors are decreased.
Misconception 4. Some students do not understand the
difference between apnea and hypoxia and that these two
features of the diving reflex are detected by separate mecha-
nisms. In the diving reflex, the voluntary apnea inhibits the
medulla respiratory center from which arise inspiratory neu-
rons that innervate the diaphragm and inspiratory intercostal
muscles. Hypoxemia, which occurs some time after the apnea
has been initiated, is detected via peripheral chemoreceptors in
the carotid body. During prolonged apnea, arterial O2 levels
decrease, whereas CO2 levels increase, and in a normal person,
these increasing arterial CO2 levels will stimulate breathing
after 1–2 min.
ѧwhen apnea (decreased oxygen) is present baroreceptors
begin to fire.
Inquiry Applications
This practical class is a methods level inquiry class, for
which students are provided with the experimental design and
questions. It serves as an introduction to experimental design,
data analysis, and the use of tests of statistical significance as
well as in the use of the Finapres recording system. Subsequent
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to this practical class, we ask students to design their own
experiments, using the Finapres technology. Thus, they rapidly
move to a guided inquiry level. In one example of an inquiry
level physiology practical class, we ask them to design and
complete an experiment to determine the acute effects of
“energy drinks” on the cardiovascular system.
This diving response practical class could easily become an
open inquiry practical class, with students generating the ques-
tions and designing the experiments. For example, students
could generate questions or experiments that are aimed at
determining the specific sensory factors that contribute to the
cardiovascular changes during the diving response. Some po-
tential questions and experiments are summarized below:
Question 1: where are the sensory receptors on the face?
Students could try exposing different parts of the face (i.e., the
forehead, nose, or cheeks) to water (see Ref. 35). For example,
a previous study (8) has demonstrated that nasal irrigation
leads to bradycardia. They could also determine whether the
diving response during facial immersion is attenuated if only
one side of the face is exposed or if a nasal clip is worn.
To determine whether exposure of the forehead to cold water
is essential for the diving response, students could compare
facial immersion with and without a face mask.
Question 2: could the activation of skin receptors on non-
facial parts of the body contribute to the diving response?
Students could compare immersion of the face in water with
immersion of some other body part, such as a hand, arm, or
foot (20). Campbell et al. (8) compared immersion of the body
with immersion of the torso (and not the face). This could be
logistically tricky in the practical laboratory setting as a bath-
tub or swimming pool may be required! There is also the
possibility that posture (i.e., leaning over the basin vs. lying
down) could affect the response, so this could also be investi-
gated.
Question 3: does training (swimming/diving/exercise) en-
hance the diving response and/or remove the anticipatory
response? The bradycardia with breath holding is smaller in
untrained than trained breath-hold divers, suggesting that there
is a training effect (13, 36). Students could divide the practical
class into swimmers/divers versus nonswimmers/divers and
test the effect of swimming/diving experience on the diving
response. They could also take into account the fitness level of
the participant (i.e., trained athletes vs. students who do no
regular exercise).
Question 4: are facial sensory receptors sensitive to tem-
perature or (water) pressure? The temperature of the ambient
air can also alter diving bradycardia (33). To dissect out the
relative contributions of the pressure or temperature of the
water on the diving response, students could alter the room
temperature. If you are fortunate, you may have access to a
temperature-controlled room. Students could also use “dry”
face cloths of known temperatures.
ACKNOWLEDGMENTS
This practical laboratory class would not be possible without the technical
support of the physiology practical laboratory staff (Scott Clarke, Rebecca
Flower, Svetlana Stanojkovic, Kushani Weerakoon, Kavitha Yattukulan). The
funding to purchase the Finapres systems was provided by the Office of the
DVC Education.
DISCLOSURES
No conflicts of interest, financial or otherwise, are declared by the author(s).

STIMULATING THE DIVING REFLEX IN HUMANS
AUTHOR CONTRIBUTIONS
Author contributions: J.K.C., K.M.D., R.G.E., and Y.H. conception and
design of research; J.K.C. and K.M.D. analyzed data; J.K.C. and R.G.E.
interpreted results of experiments; J.K.C. and K.M.D. prepared figures; J.K.C.
and Y.H. drafted manuscript; J.K.C., K.M.D., R.G.E., and Y.H. edited and
revised manuscript; J.K.C., K.M.D., R.G.E., and Y.H. approved final version
of manuscript; K.M.D. and R.G.E. performed experiments.
REFERENCES
1. AD Instruments. AD Instruments homepage (online). http://www.adin-
struments.com [18 September 2014].
2. Andersson JPA, Liner MH, Fredsted A, Schagatay EKA. Cardiovas-
cular and respiratory responses to apneas with and without face immersion
in exercising humans. J Appl Physiol 96: 1005–1010, 2004.
3. Andersson JP, Liner MH, Runow E, Schagatay EK. Diving response
and arterial oxygen saturation during apnoea and exercise in breath-hold
diver. J Appl Physiol 93: 882– 886, 2002.
4. Andersson JP, Schagatay EK. Effects of lung volume and involuntary
breathing movements on the human diving response. Eur J Appl Physiol
77: 19 –24, 1998.
5. Angell-James JE, Daly MB. Cardiovascular responses in apnoeic as-
phyxia: role of arterial chemoreceptors and the modification of their
effects by a pulmonary vagal inflation reflex. J Physiol 201: 87–104, 1969.
6. BIOPAC Systems. BIOPAC Systems hompage (online). http://www.
biopac.com/ [18 September 2014].
7. Blix AS, Folkow B. Cardiovascular adjustments to diving in mammals
and birds. Comprehens Physiol: 917–945, 2011.
8. Campbell LB, Gooden BA, Horowitz JD. Cardiovascular responses to
partial and total immersion in man. J Physiol 202: 239 –250, 1969.
9. Drummond GB, Tom BDM. Statistics, probability, significance, likeli-
hood: words mean what we define them to mean. Adv Physiol Educ 35:
361–364, 2011.
10. Drummond GB, Vowler SL. Not different is not the same as the same:
how can we tell? Adv Physiol Educ 36: 261–264, 2012.
11. Elstad M, Nådland IH, Toska K, Walløe L. Stroke volume decreases
during mild dynamic and static exercise in supine humans. Acta Physiol
(Oxf) 195: 289 –300, 2009.
12. Fagiu J, Sundlöf G. The diving response in man: effects on sympathetic
activity in muscle and skin nerve fascicles. J Physiol 377: 429 – 443, 1986.
13. Ferretti G. Extreme human breath-hold diving. Eur J Appl Physiol 42:
254 –271, 2001.
14. Finapres Medical Systems. Finometer MIDI (online). http://www.
finapres.com/Products/Finometer-MIDI [18 September 2014].
15. Folgering H, Olivier O. The diving response depresses ventilation in
man. Bull Eur Physiopath Respir 2: 143–147, 1985.
16. Fortin J, Marte W, Grüllenberger R, Hacker A, Habenbacher W,
Heller A, Wagner Ch Wach P, Skrabal F. Continuous non-invasive
blood pressure monitoring using concentrically interlocking control loops.
Comp Biol Med 36: 941–995, 2006.
17. Foster GE, Sheel AW. The human diving response, its function, and its
control. Scand J Med Sci Sports 15: 3–12, 2005.
Advances in Physiology Education • doi:10.1152/advan.00125.2013 • http://advan.physiology.org
Downloaded from www.physiology.org/journal/advances (186.141.138.240) on October 21, 2019.
Sourcebook of Laboratory Activities in Physiology
365
18. GraphPad Software. GraphPad Prism (online). http://www.graphpad.
com/scientific-software/prism/ [18 September 2014].
19. Hayashi N, Ishiharam M, Tanaka A, Asumi T, Yoshida T. Face
immersion increases vagal activity as assessed by heart rate variability.
Eur J Appl Physiol 76: 394 –399, 1997.
20. Hiebert SM, Burch E. Simulated human diving and heart rate: making
the most of the diving response as a laboratory exercise. Adv Physiol Educ
27: 130 –145, 2003.
21. Hodgson Y, Choate JK. Continuous and non-invasive recording of
cardiovascular parameters with the Finapres finger cuff enhances under-
graduate student understanding of physiology. Adv Physiol Educ 36:
20 –26, 2012.
22. Irving L, Scholander PF, Grinnel SW. The respiration of the porpoise,
Tursiops truncatus. J Cell Comp Physiol 17: 145–167, 1941.
23. Lindholm P, Lundgren CEG. The physiology and pathophysiology of
human breath-hold diving. J Appl Physiol 106: 284 –292, 2009.
24. Marabotti C, Scalzini A, Cialoni D, Passera M, L’Abbate A, Bedini R.
Cardiac changes induced by immersion and breath-hold diving in humans.
J Appl Physiol 106: 2293–297, 2009.
25. McAllen RM, Salo LM, Paton JFR, Pickering AE. Processing of central
and reflex vagal drives by rat cardiac ganglion neurons: an intracellular
analysis. J Physiol 589: 5801–5818, 2011.
26. Omron. Omron Blood Pressure Monitors (online). http://omronhealthcare.
com.au/omron-blood-pressure-monitors/ [18 September 2014].
27. Panneton WM. The mammalian diving response: an enigmatic reflex to
preserve life? Physiology 28: 284 –297, 2013.
28. Perini R, Gheza A, Moia C, Sponsiello N, Ferretti G. Cardiovascular
time courses during prolonged immersed static apnoea. Eur J Appl Physiol
110: 277–283, 2010.
29. PhysioFlow. PhysioFlow (online). http://www.physioflow.com/ [18 Sep-
tember 2014].
30. Polar World. Polar World (online). http://www.polarworld.com.au/col-
lections/heart-rate-monitors [18 September 2014].
31. R. The R Project for Statistical Computing (online). http://www.r-project.
org/ [18 September 2014].
32. Rozloznik M, Paton JF, Dutschmann M. Repetitive paired stimulation
of nasotrigeminal and peripheral chemoreceptor afferents cause progres-
sive potentiation of the diving bradycardia. Am J Physiol Regul Integr
Comp Physiol 296: R80 –R87, 2009.
33. Schagaty E, Holm B. Effects of water and ambient air temperatures on
human diving bradycardia. Eur J Appl Physiol 73: 1– 6, 1996.
34. Schelegle ES, Green JF. An overview of the anatomy and physiology of
slowly adapting pulmonary stretch receptors. Res Physiol 125: 17–31,
2001.
35. Schuitema K, Holm B. The role of different facial areas in eliciting
human diving bradycardia. Acta Physiol Scand 132: 119 –120, 1988.
36. Tocco F, Crisafulli A, Melis F, Porru C, Pittau G, Milia R, Concu A.
Cardiovascular adjustments in breath-hold diving: comparison between
divers and non-divers in simulated dynamic apnoea. Eur J Appl Physiol
112: 543–554, 2011.
37. Yale SH. Antonio Maria Valsalva (1666 –1723). Clin Med Res 3: 35–38,
2005.