The heart rates of rats aged 4, 7, and 12 days were recorded for control (CG), undernourished
(UG), control-handled (CHG), and undernourished-handled (UHG) groups after exposure to amyl
acetate and menthol. Pregnant dams were undernourished and received 50% of a balanced diet
from gestational day (G) 6 to G12, 60% from G13G18, and 100% from G19G21. On postpartum
days 012, pups remained for 12 h with a normal and 12 h with a nipple-ligated dam. The pups
were handled 5 min daily on days 112 of age to ameliorate the stress from undernourishment.
Basal heart rate, heart rate habituation, and the discrimination to a second odor were evaluated.
Body weight of UG and UHG rats was significantly reduced. Basal heart rate gradually increased
with age, especially in the CHG, without changes in the UHG; habituation was present and odor
discrimination was modified by the diet and handling. These data indicate that olfactory
discrimination is present at birth, and undernourishment does not impair but handling modifies its
functionality.
Keywords: undernourishment, handling, olfactory discrimination, habituation, rats
Introduction
In the nest environment, newborn rats may respond to
odors of the mother and nesting cage by reducing their
motor activity or by approaching their mother in a
maze in preference to either a non-lactating female or
a male. Olfactory experience may be critical for
suckling, protection, and warmth; later in development when walking emerges, the olfactory channel
plays a critical role as a basis to refine odor
discrimination and individual or species recognition
by pairing an olfactory stimulus with tactile, electrical
Correspondence to: M. Salas PhD, Departmento de Neurobiologa del
Desarrollo y Neurofisiologa, Instituto de Neurobiologa, Universidad
Nacional Autnoma de Mxico, Campus UNAM Juriquilla, Quertaro,
Qro. 76230, Mxico
Tel: +52 555623 4059; Fax: +52 555623 4001;
E-mail: masal@servidor.unam.mx
Received 11 October 2009, revised manuscript accepted 14 February 2010
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Statistical analysis
Experimental data were analyzed with the Systat
Statistical Package v7.0. To compare score differences
between the various experimental groups on physical
growth and the basal heart beat, heart beat habituation
to different odors, and olfactory cross-discrimination
between different odors, the following separate statistical
analyses were used: (i) body weight score differences were
compared in a four-way ANOVA, 2 (dietary conditions)
2 (handling condition) 3 (ages) 2 (sexes); (ii) the
basal heart rate values taken during the last 10 s before
odor presentation in each experimental group were
compared in a four-way ANOVA, 2 (dietary conditions)
2 (handling condition) 3 (ages) 2 (sexes); (iii) the
heart beat habituations to each odor were compared in a
four-way ANOVA, 2 (dietary conditions) 2 (handling
condition) 3 (ages) 2 (sexes); and (iv) score differences in olfactory cross-discrimination were compared
with a four-way ANOVA, 2 (dietary conditions) 2
(handling condition) 3 (ages) 2 (sexes). Additionally,
post hoc Fisher, least square differences (LSD) tests were
conducted for significant differences between groups at
each developmental age. A probability of 0.05 was
considered statistically significant.
Results
Effects of the age, nutritional and handling schedules
on the body weight
As shown in Figure 2, the body weight of subjects
gradually increased with age. The ANOVA comparisons
yielded significant affects of the dietary conditions (F(1,72)
= 542.80; P < 0.0001), handling condition (F(1,72) = 4.93;
P < 0.02), and age (F(2,72) = 736.02; P < 0.0001), but not
of sex (F(1,72) = 0.41; P = 0.52). Moreover, there were
significant interactions between diet and age (F(2,72) =
53.14; P < 0.0001), age and handling condition (F(2,72) =
13.36; P < 0.0001), and sex and handling condition (F(1,72)
= 5.33; P < 0.02). Post hoc comparisons at each developmental age showed significant body weight reductions (P
< 0.05) in the UG and UHG groups compared to their
controls. Furthermore, the UHG exhibited consistently
lower body weights compared to the CHG of rats.
Animals of the CG and CHG did not differ significantly
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Figure 4 Habituation of the heart beat response to amyl acetate (Odor 1) or menthol (Odor 2) in 4, 7, and 12-day-old rats in
non-handled and handled groups. The cardiac response to both odors initiates with an accelerated response
followed by a gradual deceleration except in the UHG at 4 and 12 days of age. *S1 vs S3, **S2 vs S3, S1 vs S2,
indicate significant differences (P < 0.05)
Discussion
The data showed that body weight was consistently
reduced in the UG and UHG rats when compared to
their controls, and that the body weight of UHG pups
was significantly reduced at 4 and 7 days of age, with a
small increase at postnatal day 12 when compared to
the UG of pups; there were differences between the
CG and CHG animals. The current findings agree
with previous studies showing that perinatal
undernourishment consistently interferes with the
physical development of newborns and adults, and
that food deprivation, which is usually associated with
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Conclusions
The olfactory discrimination between maternal and
novel, non-maternal odors is already functional
during the first 12 postnatal days in both UG and
CHG rats. By contrast, it was inconsistent in the CG
and UHG pups suggesting that, during this period of
life, reciprocal granule and mitral inhibitory processes
are emerging, and undernutrition associated to
handling and possible altered attentional mechanisms
interfere the olfactory discrimination by unclear
mechanisms.9,65,71 Undernourishment and handling
can increase this sensory ability, but the combination
of the two factors as an unexpected effect is consistent
with the idea that ontogenetic changes of the
underlying mechanisms are involved. However,
further studies are needed to determine the
neurophysiological levels altered by the treatments.
Acknowledgements
This work was partly supported by DGAPA/UNAM,
IN210903, IN207307. CONACyT No. 184899 and
DGAPA/UNAM No. 50410914 to MR. The authors
thank Dr Dorothy D. Pless for editorial assistance and
P. Galarza and R. Silva for collecting data.
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