331

REVIEW ARTICLE
Sperm atogenesis and Aging in the Human
LARRY JOHNSON
From the Department of Cell Biology and
Anatomy, the University of Texas Health
Science Center at Dallas,
Dallas, Texas
Key words: aging, human, spermatogenesis, sperm pro-
duction, germ cell degeneration.
J Androl 1986; 7:331-354.
The effect of age on human organs is becoming
more important as a higher percentage of people live
longer. The production of sufficient fertile spermat-
ozoa by the testis of aging men is increasingly impor-
tant as couples wait until later in life to have children
and as more aging men are available to become par-
ents. Furthermore, knowledge of testicular function
in aging men may be important in disorders such as
benign prostatic hyperplasia and impotence.
Reprint requests: Larry Johnson, Ph.D., Department of Cell
Biology, the University of Texas Health Center, 5323 Harry
Hines Boulevard, Dallas, Texas 75235.
M ost of our original studies reviewed here were supported in
part by NIH Grants AG 2260 (W . B. Neaves, P.!.) and HD 16773
(L. Johnson, P.!,) and United States Air Force contract C-0616 (R.
M . Lebovitz, P.!.).
Submitted for publication April 2, 1986; accepted for publica-
tion M ay 20, 1986.
The consensus of reports on the endocrine func-
tion of the testis in older men is that testosterone
production is reduced with age (Horton et al, 1975;
Ishimara et al, 1977; Harman and Tsitouras, 1980;
Sparrow et al, 1980; Dai et al, 1981; Harman et al,
1982; Yoshida et al, 1982; W inters and Troen, 1982;
Bremner et al, 1983; Davidson et al, 1983; Johnson et
al, 1984c; Neaves et al, 1984; W arner et al, 1985). In a
summary of 14 recent papers based on 417 old and
384 young adult men, Neaves et al (1984) found that
serum testosterone concentration was 14% lower for
the older men. Furthermore, many researchers
(Harman and Tsitouras, 1980; Nankin et al, 1981;
Harman et al, 1982; M urono et at, 1982; W inters and
Troen, 1982) agree that the testis has a reduced
ability to secrete testosterone in response to gonado-
tropin stimulation in aged men. Although the testis
in older men has been studied (Blum, 1936; M acLeod
and Gold, 1953; M olnar, 1965; Sasano and Ichijo,
1969; Bishop, 1970; Homonnai et al, 1982; Nieschlag
et al, 1982), only recently have the quantitative
aspects of spermatogenesis, expressed in terms of
sperm production rates, been evaluated in aging men
(Amann, 1981; Johnson et al, 1984b, 1984c, 1984d;
1986b; Neaves et al, 1984).
The quantitative studies revealed that daily sperm
production is significantly lower in older (50-80
years) than in younger (20-48 years) men with sim-
ilar testicular weights (Johnson et al, 1984b) or lower
332 Journal of Andrology . November/December 1986 Vol. 7
Fig. 1. Scanning electron micrograph of testicular parenchyma in the stallion. Seminiferous tubules (ST) and tails of developing
spermatids (TS) are evident. Bar length is equal to 50 m. (Johnson et al, 1978).
testicular weights (Neaves, et al, 1984; Johnson et al,
1984c). M oreover, using novel techniques, Johnson
et al (1981, 1983) determined the steps in spermato-
genesis where the age-related reduction in daily
sperm production occurs (Johnson et al, 1984b,
1984c; Johnson and Neaves, 1986). Likewise, age-
related differences in characteristics of seminiferous
tubules (Johnson et al, 1986b), Leydig cells (Kaler and
Neaves, 1978; Neaves et al, 1984), non-Leydig inter-
stitial cells (Neaves et al, 1985), myoid cells (Johnson
et al, 1986b) and Sertoli cells (Johnson et al, 1984d)
have been detected. Age-related changes in spermato-
genesis also occur in bulls (M ilk M arketing Board,
1950, 1967), rabbits (Ewing et al, 1972), mice (Gosden
et al, 1982), rats (Purvis et al, 1980; Johnson and
Neaves, 1983), and horses (Johnson and Neaves,
1981; Johnson and Thompson, 1983).
Evaluation of Spermatogenesis
in the Human Testis
The Journal of Andrology published an excellent
review on the evaluation of spermatogenesis based
on seminal characteristics (Amann, 1981). To pre-
vent duplication, this review considers only evalua-
tion of spermatogenesis by examination of testicular
tissue.
Human spermatogenesis has been evaluated by
the general appearance of seminiferous tubules (Bas-
corn and Osterud, 1925; Nelson, 1953; M annion and
Cottrell, 1961; Johnsen, 1970; Agger and Johnsen,
1978). The general appearance of seminiferous tu-
bules, even as observed by scanning electron micros-
copy (Johnson et al, 1978), can predict only an
impression of the activity of spermatogenesis (Fig. 1).
Likewise, the general appearance of tubules will not
allow detection of minor changes in spermatogene-
sis.
Differential cell counts have been employed to
quantitatively evaluate spermatogenesis in humans
(Roosen-Runge, 1956; M ancini et al, 1965). Differen-
tial cell counts have been expressed as the number of
germ cells per unit length of seminiferous tubular
circumference (Steinberger and Tjioe, 1968; Zuker-
man et al, 1978), number of germ cells per Sertoli cell
No.6 SPERM ATOGENESIS AND AGING IN THE HUM ANS Johnson
333
(Rowley and Heller, 1971; Skakkebaek and Heller,
1973; Zukerman et at, 1978), or number of germ cells
per tubule cross section (Silber and Rodriguez-Rigau,
1981; Silber, 1984). Results obtained by each of these
approaches were positively correlated with the con-
centration of spermatozoa in ejaculated semen
(Zukerman et at, 1978; Silber, 1984). Counting the
number of spermatids with elongated nuclei per tu-
bular cross section in testicular biopsies is particu-
larly suited for the clinical setting, where ease of cell
recognition, low time requirement, and good correla-
tions with sperm concentration in the ejaculate are
important (Silber, 1984).
Human spermatogenesis also has been expressed
in terms of daily sperm production. Daily sperm pro-
duction, a quantitative expression of spermatogene-
sis, is the total number of spermatozoa produced per
day (Kennelly and Foote, 1964; Amann 1970a). Daily
sperm production can be measured by cannulation of
the rete testis (Lino and Barden, 1972; Amann et al,
1974) or calculated from the number of testicular
germ cells of a given type corrected for the theoreti-
cal yield and life span of the cell types counted
(Amann, 1970a).
Clermont (1963) described the cellular associations
of germ cells constituting the six different stages of
the cycle of the seminiferous epithelium in humans.
This classification scheme still is used today. The
kinetics of spermatogenesis in humans were revealed
by autoradiographic studies in which the duration of
the cycle of the seminiferous epithelium was deter-
mined (Heller and Clermont, 1964). By adding the
IDaily sperm production is the total number of spermatozoa pro-
duced by paired testes each day. This is usually calculated from
the number of spermatids in the testis or the number of sperma-
tozoa leaving the testis (Amann, 1970a).
Potential daily sperm product ion is the potential total number of
spermatozoa produced by paired testes each day, assuming no
significant degeneration subsequent to the specific cell type being
counted. Specific cell types counted may include type B spermato-
gonia, primary spermatocytes, or secondary spermatocytes (John-
son et al, 1983).
Efficiency of spermatogenesis is a relative measure of efficiency of
sperm production obtained by dividing daily sperm production or
potential daily sperm production by the paired parenchymal
weight. This value facilitates comparisons among species that
have varied testicular weights (Amann, 1970a) and allows com-
parisons of sperm production rates at different matrrational
steps during spermatogenesis (Johnson et al, 1983, 1984a; John-
son, 1985).
Daily sperm oulpul is the total number of spermatozoa released
daily by paired testes. This can be estimated by the number of
spermatozoa obtained by cannulation of the excurrent ducts of
the testis, spermatozoa recovered from the urine or the ejaculate
(Amann, 1970a). Daily sperm output has been reevaluated in
humans following stabilization of extragonadal sperm reserves
(Johnson, 1982).
durations of each stage of the cycle in which a given
cell type occurs, the life span of that cell type can be
determined. Given the life span and theoretical yield
of specific cell types, sperm production rate obtained
by counting numbers of germ cells in both testes can
be expressed on a daily basis (Kennelly and Foote,
1964; Swierstra, 1967; Amann and Lambiase, 1969;
Amann, 1970a). The theoretical yield is calculated by
2, where n equals the number of cell divisions
between a specific cell type and spermatozoa. If the
theoretical yield is above one (the value for sperma-
tids), the expression of sperm production is called
potential daily sperm production (Johnson et al,
1983).
It is only recently that daily sperm production or
potential daily sperm production has been estimated
for humans (Amann and Howards, 1980; Johnson et
al, 1980a, 1980b; 1981, 1983, 1984b, 1984c, 1984d;
1986b; Amann, 1981; Neaves et at, 1984; Johnson
and Neaves, 1986). Calculating daily sperm produc-
tion per g decapsulated testicular parenchyma facili-
tates comparisons among species as it is a measure of
efficiency of spermatogenesis (of sperm production)
regardless of testicular size (Amann, 1970a; Amann
et al, 1976). Humans (Amann, 1981; Johnson et at,
1981) have a lower efficiency (6 X 106/g) of spermato-
genesis than other species including non-human
primates (23 X 106/g), rats (20-24 X 106/g), rabbits
(26-28 X 106/g) and horses (19 X 106/g) (Amann,
1970a; 1981; Amann et at, 1976; Robb et al, 1978;
Johnson et at, 1980a, 1984a; Johnson and Neaves,
1983; Johnson and Thompson, 1983; Lebovitz and
Johnson, 1983; Johnson, 1985a).
The long duration of spermatogenesis (74 days) in
humans (Heller and Clermont, 1964) and a lower
number of germ cells per unit volume of parenchyma
(density) compared with other more efficient species
(Figs. 2 and 3) contribute to the low efficiency of
spermatogenesis in humans. The human testis also
has lower percentages of seminiferous tubule, sem-
iniferous epithelium, and germ cell nuclei than the
testis from rats or horses (Fig. 2). Using spermatids
with a spherical nucleus (round spermatids) to depict
the density of germ cells in the stage of the cycle in
which spermiation occurs, the human testis is com-
pared with the rat and horse testis (Fig. 3). The
reduced density of spermatids with a spherical nu-
cleus in the human testis compared with other spe-
cies can be seen in regions of tubules where these
germ cells are relatively abundant. Unlike seminifer-
ous tubules from rats or horses, tubules in humans
are not consistent in the abundance of germ cells
UsEt.W EROUS
TUBULES
,-
sEM*EROUS ROUM SPERMATI
EPm#{128}W M MJc1El
RAT HORSE HUMAN
N=3 N3 N=3
334 Journal of Andrology November/December 1986 Vol. 7
> -
I
0
z
LU
0
U-
0
I-
z
Ui
0
LU
0
1
Fig. 2. Comparison of the relative percentage of testicular parenchyma occupied by seminiferous tubules, seminiferous epithelium, and
spherical nuclei of round spermatids (Golgi and cap phases) in rats, horses, and humans. (Johnson et al, 1980; Johnson, 1985a).
within a specific stage of the cycle. Figure 4 illustrates
the relative paucity of spermatids with a spherical
nucleus in several seminiferous tubules at the same
stage of the cycle (that of spermiation) in the same
human testis as in Figure 3. Other stages of the cycle
often have missing generations of germ cells charac-
teristic of that specific stage. Low numbers and the
lack of all generations of germ cells in each stage of
the cycle contribute to the relatively low number of
germ cells in the human testis, which in turn con-
tributes to the low efficiency of spermatogenesis. It is
unclear why the human testis differs from that of
other species in the composition and duration of sper-
matogenesis.
Values for daily sperm production for a species are
similar to the number of spermatozoa ejaculated
daily (daily sperm output; Amann, 1970a) for that
species. Daily sperm output is the total number of
spermatozoa released daily by paired testes (Amann,
1970a; 1981). This is true for boars (Swierstra, 1968),
rabbits (Amann, 1970b), bulls (Amann et al, 1974),
horses (Gebauer et al, 1974) and dogs (Olar et al,
1983). Even in humans (Fig. 5), daily sperm produc-
tion is similar to daily sperm output (Johnson, 1982).
In the latter comparison, daily sperm output was
estimated in nine men (32 1 year) by averaging the
number of spermatozoa in the last three of five daily
ejaculates. The first ejaculates were excluded since
the extragonadal sperm reserves had to be stabilized
by depletion of epididymal spermatozoa (Amann,
1970a). It was found that two daily ejaculates were
sufficient to stabilize the extragonadal sperm reserves
in men following depletion (frequent ejaculations) or
build up (sexual rest for 6 days) of these epididymal
sperm reserves (Johnson, 1982). Daily sperm produc-
tion was estimated from 10 men of similar age (35 3
years) from counts of spermatids in the testis (John-
son et al, 1981). Although daily sperm production is
inherently higher than daily sperm output (Amann,
1970a), the similarity between them gives validity to
the concept of daily sperm production as a direct
measure of spermatogenesis in humans (Johnson et
al, 1981; Johnson, 1982).
Degeneration of germ cells during spermatogene-
sis has been estimated by comparing daily sperm
production or potential daily sperm production based
on given germ cell types at different maturational
steps in spermatogenesis in bulls (Kennelly and
Fig. 3. Comparison of seminiferous epithelium in the a) rat, b) horse, and c) human in the stage of the cycle when spermiation occurs.
Golgi and cap phase spermatids with spherical nuclei called round spermatids (RS) are more abundant in the rat and horse than in the
human. In the case of the human, this tubule was selected for the relative abundance of spermatids with spherical nuclei in relation to other
tubules in the same stage of the cycle (compare with Fig. 4). Bar length equals 10 sm.
#{149}
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1/
-
p #{149}
Fig. 4. Comparison of seminiferous epithelium and Leydig cells from the same human testis depicted in Fig. 3. Specimens were a) fixed
within 20 mm of castration orb) fixed after 12 hours of incubation at room temperature in an air-tight vial. Quality of fixation of germ cells
(especially pachytene old primary spermatocytes lOPS]) is reduced with incubation time. The size of cell boundaries and nuclei of these
germ cells do not appear to be affected. However, the cytoplasm is contracted from the cell boundary. Leydig cells (LC) appear to be less
affected with incubation time than do primary spermatocytes. Bar length equals 10 zm.
I
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337
UYJGER ADILT MEN
P1-89
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OLD#{128}R ADILT M EN
p1-43
RIG4T TESTIS
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W EIGHT OF TUNICA PERCENT TUNIC
ALBUGINEA OF W HOLE TESTIS
No.6
200- ___
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DAILY SPERM PRODUCTION VS
DAILY SPERM OUTPUT IN HUMANS
Fig. 5. Comparison between daily sperm production estimated
by evaluation of testes in 10 men aged 26 to 53 yrs (Johnson eta!.,
1981) and daily sperm output determined by counting the
number of spermatozoa in the last 3 of 5 daily ejaculations in 9
similarly aged men (Johnson, 1982).
Foote, 1964), rats (Johnson et at, 1984a), horses
(Johnson, 1985a) and humans (Barr et al, 1971; John-
son et al, 1981, 1983, 1984c; Johnson and Neaves,
1986).
Age-related Differences
in the Human Testis
Our research has used autopsy specimens from
individuals who died of heart attack or traumatic
injury (i.e. automobile accident or gun shot wound).
Study subjects included only men with no history of
serious illness (other than coronary arteriosclerosis)
and no extended hospitalization prior to death.
Details on the subjects apparent good health prior to
death, ethnic origin, cause of death, body weight,
blood alcohol concentration, and ratio of body weight
to height have been described (Neaves et al, 1984;
Johnson et al, 1986b). Postmortem changes reduce
the quality of fixation and cause contraction of the
cytoplasm in primary spermatocytes (Fig. 4). How-
ever, we have been unable to show an effect of post-
mortem time on daily sperm production (Johnson et
al, 1984b).
Age-related differences in testicular weight and
testicular volume are inconsistent features of aging
in humans. However, testicular weight does not
increase after age 20 to 30 years in humans. Compar-
ing data for 89 men 21 to 50 years old and 43 men 51
Fig. 6. Effect of age on the weight of the tunica albuginea and
the percent tunic of the whole testis in younger adult (21-50 yr)
and older adult (51-80) men. (Johnson et al, 1984b).
to 80 years old, testicular weights were similar for
both the right (18.9 0.5 and 19.2 0.9 g, respec-
tively) or left (17.2 0.5 and 17.5 0.9 g, respec-
tively) testis (Johnson et al, 1984b). This is consistent
with similar testicular volumes reported for young
and old men (Kothari and Gupta, 1974). However,
the weight of the tunica albuginea and the percent-
age of total testicular weight represented by the tu-
nica albuginea were greater (P < 0.01) for the older
men (Fig. 6). Subsequent studies on 15 men aged 20
to 48 years and 15 men aged 50 to 76 years revealed
an age-related difference (P< 0.01) in the volume of
the testicular parenchyma (40.2 2.3 ml and 28.2
2.4 ml, respectively) per man (Neaves et al, 1984).
The age-related increase in the weight of the tunica
sometimes may be sufficient to mask differences in
parenchymal weight. Our recent study (Johnson et
al., 1986b), based on 28 men 20 to 48 years old and 28
men 50 to 90 years old, revealed age-related differ-
ences (P< 0.01) in both testicular weight and paren-
chymal weight (Table i). Age-related differences in
parenchymal weights in the left, right, or paired tes-
tis in younger adult (20-48 years) and older adult men
(50-90 years) are illustrated in Figure 7. Others have
reported that changes in testicular volume are not a
consistent characteristic of aging (Sterns et al, 1974;
Nieschlag et al, 1982).
The composition of the human testis is modified
with age. Comparing 28 men 20 to 48 years and 28
SPERM ATOGENESIS AND AGING IN THE HUM AN. Johnson
U)
.4
U)
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=
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z
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a-
D YOU*GER ADCLT M EN OLDER ADCLT MEN
N50 N44
40- ..L
.1
20-
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L P1-28
OLD#{128}R
L:I P1-28
338 Journal of Andrology . November/December 1986 Vol. 7
SEMINIFEROUS
TUBULES
SEMINIFEROUS
EPITHELIUM
Fig. 7. Effect of age on parenchymal
(decapsulated testicular tissue) weight of
the left, right, or total (paired) testes in
younger adult (20-48 yrs) and older adult
(50-90 yrs) men. (Johnson et al, 1984b).
men 50 to 90 years old (Johnson et al, 1986b), the
percentage of seminiferous epithelium was reduced
(P < 0.01) in older adult men (Fig. 8). However, no
significant difference was found in volume density
(percentage of parenchymal volume occupied by a
given component) of seminiferous tubules or tubular
lumen. The volume density of boundary tissue
(myoid cells and associated extracellular components)
increased (P< 0.01) with age (Fig. 9), but the volume
density of myoid cells in the boundary tissue was
similar in younger and older men (Johnson et al.,
1986b). Partly due to age-related differences in pa-
renchymal weight, the volume of seminiferous tu-
bules per man was less (P < 0.01) in the older men
(Table 1). Tubular diameter was similar but tubular
length per man was reduced (P< 0.01) in older adult
men (Fig. 10). The thickness of the boundary tissue,
percent boundary tissue in the parenchyma, and
number of myoid cells per cross section increased (P
<0.01) in older adult men (Fig. 9). W here there was
tubule hyalinization in the human testis, an increased
deposition of collagen fibers was reported (Soder-
stom, 1986). The age-related thickening of the boun-
dary tissue in our study was not due to an increase in
boundary tissue itself, but resulted from a reduction
(P< 0.01) in tubule length per man without a reduc-
tion in the total volume of boundary tissue per man
(Table 1). These conclusions, based on examination
>-
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z
Lii
0
Li
0
H-
z
Lii
0
lx
Lii
0
COMPOSITION (VOLUME DENSITY)
Fig. 8. Effect of age on the percentage
of the testis occupied by seminiferous
tubules (P> 0.05) and seminiferous epi-
thelium (P < 0.01) in younger adult (20-
48 yrs) and older adult (50-90 yrs) men.
(Johnson et al, 1986b).
UYOUNGER
3OLDER
2e
1
BCUIARY TISS.E PEREHT M1ER OF KYO SELLS
T1IOPESS U14) BO(1IARY 715511 PER CROSS SECTION
No.6 SPERM ATOGENESIS AND AGING IN THE HUM AN . Johnson 339
* From Johnson et al, 1986b.
tMean SEM.
Fig. 9. Effect of age on boundary tissue
thickness (P < 0.01), percent boundary
tissue (P < 0.01), and number of myoid
cells per cross section (P < 0.01) in
younger adult (20-48 yrs) and older adult
(50-90 yrs) men. (Johnson et al, 1986b).
TABLE 1. Age-related Variation in Sem iniferous Tubules and Daily S perm Production in Men*
Testicular Component
Age Group
Significance
20-48 yr
(n = 28)
50-90 yr
(n = 28)
W eight
Body (kg)
Paired testes (g)
Paired testicular parenchyma (g)
Volume per man (ml)
Seminiferous tubule
Boundary tissue
Lumen
Seminiferous epithelium
77.6 3.3t
46.8 1.8
40.7 1.7
23.81.0
3.5 0.2
3.5 0.4
16.9 0.7
79.2 3.7
37.8 2.0
31.4 1.8
18.01.2
3.4 0.2
3.2 0.4
11.40.9
NS
P <0.01
P <0.01
P<O.O1
NS
NS
P <0.01
Myoid cell
Volume density of boundary tissue (%) 32.1 2.2 28.9 2.3 NS
Total volume per man (iiI) 1109 51 970 51 NS
Volume of average individual cell (Il) 1197 54 1322 66 NS
Number of cells
Sertoli cells (106)
Per g parenchyma
Perman
Spermatids with round nucleus
Per g parenchyma
Perman
Spermatid with round nucleus to Sertoli cell ratio
23.9 1.1
97761
53.5 3.4
2226194
2.3 0.2
16.9 1.0
55151
34.1 2.9
1075104
2.1 0.2
P< 0.01
P<0.O1
P <0.01
P<O.O1
NS
Daily sperm production (106)
Per g parenchyma
Per man
6.0 0.4
250 22
3.8 0.3
121 12
P <0.01
P <0.01
UYOUNGER
P1=28
MOLDER
N28
ri#{149}
TUBULAR
LENGTH/MAN (M)
AGE, years
340 Journal of Andrology November/December 1986 Vol. 7
Degeneration of germ cells has a pivotal role in the
TUBULAR
DIAMETER (UM)
Fig. 10. Effect of age on tubular diameter (P>0.05) and tubu-
lar length per man (P < 0.01) in younger adult (20-48 yrs) and
older adult (50-90 yrs) men. (Johnson et al, 1986b).
of the whole testis instead of biopsies, are in contrast
to earlier, more subjective, studies that concluded
that the age-related thickening of boundary tissue
resulted from deposition of new layers of connective
tissue (Engle, 1942; M olnar, 1965).
Age-related changes in sperm production in hu-
mans are evident in a higher percentage of azoo-
spermia in semen from older men (Blum, 1936),
tubules devoid of spermatids (Sasano and Ichijo,
1969), tubules with dislocated type A spermatogonia
(Holstein et al, 1984), or multinucleated spermato-
cytes and spermatids (Holstein and Eckmann, 1986).
Honore (1978) noted that age-related changes in the
human testis included tubular sclerosis, focal mono-
nuclear orchitis, and dilation of the rete testis. Bishop
(1970) reported a thinning of the spermatogenic
epithelium, decreased diameter of tubules, and even-
tual obliteration of the tubular lumen in some old
men. Lower seminiferous epithelial volume in older
men is associated with lower (P < 0.01) daily sperm
production per man (Table 1; Fig. 11; Johnson et al,
1986b).
Combining the reported data from different stud-
ies (Amann and Howards, 1980; Johnson et al, 1980a,
1980b), Amann (1981) reported a trend for an age-
related difference in daily sperm production in men.
This trend has been confirmed by subsequent studies
(Johnson et al, 1984b; 1984c; 1984d; 1986b; Neaves et
al, 1984). The efficiency of spermatogenesis (daily
sperm production/gram parenchyma; Amann, 1970a)
in older men is less (P< 0.01) than in younger adult
men (Table 1). M oreover, the low efficiency of sper-
AGE vs SPERM PRODUCTION
Fig. 11. Effect of age on daily sperm production per gram
parenchyma (efficiency of spermatogenesis) in 132 men. Regres-
sion line (y = 7.7-0.065X; P = -0.33) is drawn with the 95%
confidence limit. (Johnson et al, 1984b).
matogenesis is uniformly distributed in each testis
(Johnson et al, 1984b) and in the cranial, equatorial,
and caudal thirds of the testis of young or older men
(Johnson et al, 1984c). Comparisons of the efficiency
of spermatogenesis between the right and left testis
and amount of daily sperm production in different
regions can be seen in Figure 12. Serum concentra-
tions of LH and FSH were positively correlated with
age and negatively correlated with daily sperm pro-
duction (Johnson et al, 1984c).
Age-related differences in the number of germ
cells or daily sperm production in the testis of older
(36-month-old) rabbits (Ewing et al, 1972), mice
(Gosden et al, 1982) and rats (Johnson and Neaves,
1983) have been reported. In horses, daily sperm
production reaches its peak at about 4to 5 years of
age and remains at this level until at least 20 years of
age (Johnson and Neaves, 1981; Johnson and Thomp-
son, 1983). W hat happens to sperm production after
20years of age in stallions remains unknown. The
fact that some males retain their fertility even with
the advancement of age is indicated by fertility in
bulls 19 years of age (Bishop, 1970) and a successful
paternity in a 94-year-old man (Seymour et al, 1935).
Degeneration of Germ Cells
During Spermatogenesis
N=1O YOUNG ADULT MEN
Fig. 12. Daily sperm production per g
parenchyma in the cranial, equatorial,
and caudal regions of paired testes from
men aged 20 to 48 yrs. (Johnson et al,
1984c).
No.6 SPERM ATOGENESIS AND AGING IN THE HUM AN. Johnson 341
CRANIAL EQUATORIAL CAUDAL
quantitative aspects of spermatogenesis (Huckins,
1978; Johnson et al, 1983; Johnson, 1985a). However,
the mechanisms of degeneration, its etiology, and
approaches for prevention remain unclear.
Three approaches have been used to quanitify the
amount of germ cell degeneration. The extent of
germ cell degeneration at different steps of spermato-
genesis has been determined by counting degenerat-
ing germ cells of different types (Oakberg, 1956;
Clermont, 1962; Russell and Clermont, 1977; Huck-
ins, 1978) and by calculating ratios of specific types of
germ cells (Roosen-Runge, 1955; 1973; Oakberg,
1956; Clermont, 1962; Amann, 1962; Barretal, 1971;
Huckins, 1978; Hochereau-de Reviers, 1981; W ing
and Christensen, 1982). The identity of degenerating
cells and the fact that degenerating cells are removed
quickly limits the usefulness of the first approach. A
third approach, which directly estimates the impact
of degeneration at given steps in spermato genesis on
daily sperm production, is to compare rates of sperm
production based on various types of developing
germ cells (Swierstra, 1967; Amann, 1970a). The last
approach (Table 2) has been employed to estimate the
effect of microwave irradiation of rats on sperm pro-
duction from type B spermatogonia to the most
mature maturation-phase spermatids (Lebovitz and
Johnson, 1983; Johnson et at, 1984a) as well as sea-
sonal differences in germ cell degeneration from type
B spermatogonia to spermatids in stallions (Johnson,
1985a). In the adult man, this approach has revealed
U,
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0
cr>-
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IX
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a-
significant degeneration of germ cells during sperma-
togenesis (Fig. 13; Table 2) and age-related differen-
ces in the types of germ cells that degenerate
(Johnson et al, 1983, 1984c). All three methods of
evaluating germ cell degeneration have revealed con-
siderable germ cell loss and have led to the concept
that degeneration of a percentage of developing
germ cells is a normal phenomenon of spermatogene-
sis (Roosen-Runge, 1955; 1973; Clermont, 1962;
Barr et al, 1971; Russell and Clermont, 1977; Huck-
ins, 1978; Johnson et at, 1983).
Germ cell degeneration has been associated with
three critical steps in spermatogenesis. During sper-
matogonial mitoses, 25% of the potential progeny
from type A spermatogonia are not produced in mice
(Oakberg, 1956), 11% are lost in Sherman rats
(Clermont, 1962), and 75% are lost in adult Sprague-
Dawley rats (Huckins, 1978). Ortavant (1958) found
greater degeneration of spermatogonia in rams fol-
lowing long day illumination. M eiotic divisions ac-
count for a 13% loss of potential production in mice
(Oakberg, 1956), 2% in Sprague-Dawley rats (Roo-
sen-Runge, 1955; Johnson et al, 1984a), 27% loss in
Sherman rats (Clermont, 1962), 25% in rabbits
(Swierstra and Foote, 1963), and 6% and 15% losses
in stallions during the breeding and non-breeding
seasons, respectively, (Johnson, 1985a). Ortavant
(1958) found 40 to 50% fewer spermatids in rams
following long day illumination. Losses due to degen-
eration during spermiogenesis are undetermined in
342 Journal of Andrology . November/December 1986 Vol. 7
TABLE 2. The Effect of Germ Cell Degeneration at Different Steps in Spermatogenesis
on Sperm Production in Adult Rats, Horses, and Humans*
Criterion
Species
Rat Horse Human
Parenchymal weight (9) 1.60 0.04 151 8 16 1
Potential daily sperm production/g parenchyma (106)t
Type B spermatogonia
Young primary spermatocytes
Old primary spermatocytes
Secondary spermatocytes
Daily sperm production/g (106)
Young spermatids
Old spermatids
20 2
20 2
24 1
-
20 1
25 1
32
21 1
22 f
-
20 1
19 1z
12 2
10 l
12 2
6 f
6 1z
Potential daily sperm production/testis (106)t
Type B spermatogonia
Young primary spermatocytes
Old primary spermatocytes
Secondary spermatocytes
Daily sperm production/testis (106)
Young spermatids
Old spermatids
32 4
33 2
37 1
-
31 1
41 1
4000 2OO
2100 20(f
2000 200z
-
1700 200
1700 2002
216 34
172 22
210 47
100 17z
96 14
*Data (mean SEM) compiled from our studies on 14 adult (>4009) rats (Lebovitz and Johnson, 1983; Johnson et al, 1984a), 28
horses in the breeding season (Johnson, 1985a), and 10 humans ages 26 to 53 years (Johnson et al, 1984c).
tPotential daily sperm production is the predicted number of spermatozoa possible when based on numbers of designated cell
types younger than spermatids.
Means in columns with different superscripts are different (P <0.05).
2Evaluated in the contralateral testis for the rat and horse.
Sherman rats (Clermont, 1962) and mice (Oakberg,
1956) and 6% or less in stallions (Johnson, 1985a).
Bulls (Amann, 1962) and adult (>400 g) Sprague-
Dawley rats (Lebovitz and Johnson, 1983; Johnson et
al, 1984a) appear to have no significant germ cell loss
during spermiogenesis.
There was no significant degeneration of germ
cells between type B spermatogonia and secondary
spermatocytes (Table 2). The lack of details on sper-
matocyte formation in humans prevents evaluation
of germ cell degeneration during proliferation of
spermatogonia. A significant 36% to 45% loss by
degeneration (Barr et al, 1971; Johnson et al, 1983)
occurs during meiotic divisions in humans. The
amount of potential loss of sperm production late in
meiosis is significantly and negatively correlated (r =
-0.86) with daily sperm production per gram paren-
chyma in humans (Johnson et at, 1983). The majority
of the germ cell degeneration detected late in meiosis
occurred during the second meiotic division (Johnson
et al, 1981, 1984c). The percentage of germ cell degen-
eration during postprophase of meiosis was not
correlated with age or serum concentrations of
either LH or FSH (Johnson et al, 1984c). It is not
known why humans have a greater degeneration
rate late in meiosis than other species (Table 2; Fig.
13). Based on the magnitude of cell loss late in meiosis
in humans (Johnson et al, 1983), daily sperm produc-
tion almost could be doubled if this loss was elimi-
nated. In adult humans, there is little degeneration of
spermatids during spermiogenesis (Table 2; Johnson
et al, 1981).
In summary, the effect of germ cell degeneration
on daily sperm production (Table 2; Fig. 13) was
negligible in adult (>400 g) rats subsequent to type B
spermatogonia, while adult horses had losses toward
the end of spermatocyte formation and younger
adult humans had great losses after the end of mei-
osis (subsequent to secondary spermatocytes).
The reasons germ cells degenerate are unclear.
Oakberg (1956) and Clermont (1962) proposed that
the degeneration of germ cells may be a mechanism
for eliminating cells with abnormal chromosomes.
However, the greater loss of certain types of sperma-
togonia and the relative constancy in the magnitude
of degeneration are not consistent with simple selec-
tion to eliminate chromosomal abnormalities (Huck-
ins, 1978). Alternatively, degeneration might be a
mechanism to limit germ cells to the number that can
be sustained by the Sertoli cell population (Huckins,
1978). Indeed, the number of type A spermatogonia
per testis has been correlated with numbers of Ser-
Fig. 13. Potential daily sperm produc-
tion per g parenchyma evaluated at dif-
ferent steps in spermatogenesis in the
rat, horse, and human. Cell types on
which potential daily sperm production
per g or daily sperm production per g was
based included: type B spermatogonia
(BS), preleptotene plus leptotene (young
primary) spermatocytes (YPS), pachytene
plus diplotene (old primary) spermato-
cytes (OPS), secondary spermatocytes
(SS), Golgi and cap phase spermatids
(young spermatids) (YS), and matura-
tion-phase spermatids (old spermatids)
(OS). Adult (>400 g) rats experienced no
significant loss during these different
steps in spermatogenesis (Lebovitz and
Johnson, 1983; Johnson et al, 1984a).
Adult horses had early losses in spermato-
genesis (end of spermatocyte formation)
with no subsequent losses (Johnson,
1985a). Younger adult humans had sig-
nificant losses during postprophase of
meiosis after secondary spermatocytes
(Johnson et al, 1984c). Specific values and
significance levels are given in Table 2.
I
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40 -
30 -
1
T
20 - f J
10 -
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RAT HORSE HUMAN
I I I I
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D YOIM aaLT 4 OLD(R DLT Il
P37 P1-34
Fig. 14. Effect of age on the number of Sertoli cells per testis in
younger adult (20-48 yrs) men and older adult (50-85 yrs) men.
(Johnson et al, 1984d).
No.6 SPERM ATOGENESIS AND AGING IN THE HUM AN. Johnson 343
toli cells in rats, rams, bulls (Hochereau-de Reviers
and Courot, 1978) and stallions (Johnson and Thomp-
son, 1983; Johnson, 1985a).
Given that the population of type A spermatogo-
nia in stallions increases two-fold in the breeding
season and that there is no offsetting increase in
degeneration between type B spermatogonia (Table
2) and spermatids, each Sertoli cell would have to
accommodate 60% more germ cells (Johnson, 1985a).
The ratio of all types of germ cells to Sertoli cells
would increase from 28:1 (Johnson, 1986) to 44:1 in
the breeding season.
Our studies with humans support the premise that
degeneration has a role in regulating the number of
germ cells to that which can be sustained by germ
cell-Sertoli cell interactions (Huckins, 1978). Age-
related reduction in daily sperm production in hu-
mans (Table 1) is associated with an age-related loss
of Sertoli cells (Fig. 14); the number of primary sper-
matocytes or spermatids per Sertoli cell was not
affected by age (Table 1; Johnson et at, 1984d).
Age-related Difference in Germ
Cell Degeneration in Humans
Details of spermatogonial division during sper-
matocyte formation are lacking for humans. Hence,
precise identification of different subtypes of sper-
matogonia that degenerate during spermatogonial
mitosis is impossible. However, three types of sper-
matogonia (Fig. 15) have been identified (Clermont,
1963), and comparisons have been made between the
numbers of each type as a function of age. Based on
the number of spermatogonia per gram parenchyma,
there was no significant difference between men
aged 20 to 48 years and men aged 52 to 90 years
(Johnson and Neaves, 1986). However, there was a
trend toward more type A dark and type A pale
spermatogonia and fewer type B spermatogonia in
the older men. W hile the number of type A dark
spermatogonia per man was not significantly less in
the older men, the numbers of type A pale per man
and type B per man were significantly less in the
older men.
OF SERTOLI CELLS PER TESTIS (MILLIONS)
200 490 690
344 Journal of Andrology - November/December 1986 Vol. 7
The age-related difference in daily sperm produc-
tion largely resulted from germ cell degeneration
during the prophase of meiosis in older adult men
(Fig. 16; Johnson and Neaves, 1986). There was no
difference (P> 0.05) in the number of young primary
spermatocytes per g parenchyma between younger
and older adult men. Only the older men had a signif-
icant reduction in potential daily sperm production
between young and old primary spermatocytes.
Hence, the degeneration of late leptotene, zygotene,
or pachytene primary spermatocytes in older adult
men was the major source of the age-related differ-
ence in daily sperm production. It should be pointed
out, however, that the number of type A pale sper-
matogonia per man as well as the number of each
subsequent cell type was less (P < 0.01) in the older
men. W hile this is consistent with some age-related
difference occurring during spermatocyte formation
in humans, the major age-related effect was during
prophase of meiosis.
The difference in numbers of pachytene primary
spermatocytes in older men was subsequently re-
flected (Fig. 13) at the level of spermatids with a
spherical nucleus (Johnson et al, 1984c, 1986b) and
maturation-phase spermatids (Johnson et al, 1984b).
Both age groups experienced significant degenera-
tion at the end of meiosis (Johnson et al, 1984c). Since
these losses were proportional between age groups,
it did not contribute to age-related differences in
daily sperm production.
The difference in the number of spermatids with a
spherical nucleus per man between younger and
older adult men (older men have 66% or less than
that of younger men; Johnson et al, 1984c) is similar
to the difference in the number of maturation-phase
spermatids per man (older 71% of younger men;
Johnson et al, 1984b). Thus, there appears to be no
age-related difference in degeneration during sper-
miogenesis in humans. W hen daily sperm production
based on spermatids with a spherical nucleus was
compared with daily sperm production based on the
number of maturation-phase spermatids or on the
total number of all types of spermatids in the same
men, no degeneration was found during spermato-
Fig. 15. Types A dark (Ad) and A pale (Ap) spermatogonia, preleptotene and leptotene primary spermatocytes, collectively called young
pimary spermatocytes (YPS), pachytene plus diplotene primary spermatocytes, collectively called old primary spermatocytes (OPS), Golgi
an i cap phase spermatids, collectively called young spermatids (YS), and Sertoli cells (SC) as observed in 1.0-him toluidine blue-stained
Epon sections of human seminiferous tubule (Stage III) following vascular perfusion with glutaraldehyde and fixation with 0s04. Bar
length is 10 zm.
E - CILT tI iW ILT tl
T
i-I
Fig. 16. a) Effect of age on the number
of various germ cells per g (P> 0.05 on all
types compared). b) Effect of age on
potential daily sperm production per g
parenchyma. Significant differences be-
tween age groups were found for old
primary spermatocytes (pachytene plus
diplotene) and young spermatids (Golgi
and cap phases). In the younger age group,
the value for young spermatids was lower
(P< 0.05) than values for other cell types.
In only the older men, there was a signifi-
cant reduction in potential daily sperm
production between young (preleptotene
and leptotene) and old primary sperma-
tocytes. It is this degeneration or loss of
late leptotene, zygotene, or pachytene
primary spermatocytes that largely ex-
plains age-related differences in daily
sperm production per g parenchyma.
(Johnson and Neaves, 1986).
ri
ADAC APaLE S
SP!R45.TOCY1U a
.1.
T
B a-a
SP46TOGOI6 P4RY
SPE4ATOCYTE5 5P4ATOCYTES
YOU.
1s
No. 6 SPERM ATOGENESIS AND AGING IN THE HUM AN . Johnson 345
I,,
z
0
-J
-J
I
I
C,
L&I
a-
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-J
-J
Lii
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z-s
2(/)
I-z
O=j
LIi>
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z
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=
-:
Z(D
LiJ
Ow
a-a-
genesis (Table 2; Fig. 13; Johnson et al, 1981).
Age-related Difference in Numbers of
Nongerminal Testicular Cells
The age-related decline in the response of the tes-
tis to hCG (Nankin et a!, 1981; M urono et a!, 1982)
may be related to the reduction in the number of
Leydig cells in older men. Negative relationships
between age and numbers or mass of Leydig cells
have been reported (Teem, 1935; Sargent and M c-
Donald, 1948; Tillinger, 1957; Harbitz, 1973; Kaler
and Neaves, 1978; Neaves et a!, 1984; Neaves and
Johnson, 1985). Since there are reductions in the
b
volume density of Leydig cells in the parenchyma,
and the volume or number of Leydig cells per gram
parenchyma is reduced in older men (Neaves et at,
1984), there is a reduction in the total number of
Leydig cells per man. In contrast, Kothari and Gupta
(1974) and Honore (1978) reported hyperplasia of
Leydig cells with advancing age in humans. Compar-
ing species, Ewing et al. (1979) found that Leydig cell
mass was not related to the amount of testosterone
produced during in vitro perfusion. However, Zirkin
et at (1980) found that the percentage of smooth
endoplasmic reticulum in the Leydig cell almost
totally explained the variation in the in vitro produc-
Fig. 17. The annual cycle of the Leydig
cell population in stallions revealed by
evaluation of each 3-month period for
one year. The numbers of Leydig cells per
g parenchyma were similar (P > 0.05)
among the four 3-month time periods.
The number of Leydig cells per testis for
May to July was higher (P < 0.05) than
August to October or February to April
and higher (PJ <I 0.01) than in November
to January. W hile the numbers of Leydig
cells per testis were similar (P > 0.05)
between February to April and August to
October and between August to October
and November to January, values for
February to April were significantly (P <
0.05) higher than those in November to
January.
U)
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Ui
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0
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0
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Id
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I
346 Journal of Andrology - November/December 1986 Vol. 7
NOV-JAN FEB-APR MAY-JU.. AUG-OCT
tion of testosterone in the testis maximally stimu-
lated with LH. In rats and horses, the numbers of
Leydig cells were not reduced in aged males (Kaler
and Neaves, 1981; Johnson and Neaves, 1981; John-
son and Thompson, 1986). In addition, the size of the
Leydig cell population cycles annually regardless of
age in adult horses (Fig. 17; Johnson and Thompson,
1986).
The number of Leydig cells is correlated (r = 0.76; P
<0.01) with daily sperm production in horses (John-
son and Thompson, 1986), and a seasonal increase in
the number of Leydig cells precedes increases in daily
sperm production (Johnson, 1985b). However, there
is no relationship between number of Leydig cells
and daily sperm production in humans (Neaves et a!,
1984; Neaves and Johnson, 1985). Even attempts to
enhance the likelihood of a significant relationship
between Leydig cell number and daily sperm produc-
tion by evaluating only older men (50-80 years), who
should have lost the ability to compensate for the
paucity of Leydig cells, failed to yield a significant
relationship in humans (Neaves and Johnson, 1985).
In humans, age-related changes in the ultrastruc-
ture of Leydig cell nuclei have been reported (Yasu-
zumi et a!, 1967), and a detailed description of Leydig
cell ultrastructure in aged men has been described
(M on et al, 1982). Earlier light microscopic studies of
Leydig cells in aging men (Lynch and Scott, 1950)
revealed decreased frequency of Leydig cells with
small lipid droplets and increased numbers of pig-
mented Leydig cells. However, the ultrastructure of
Leydig cells present in aging men can appear quite
normal (Fig. 18).
Age-related reduction in the number of Leydig
cells (Neaves et a!, 1984), was not associated with a
corresponding increase in the number of non-Leydig
interstitial cells in the same men (Neaves et a!, 1985).
Apparently, Leydig cells degenerate rather than
dedifferentiate into mesenchymal cells (Sniffen, 1950;
Neaves et a!, 1985). M yoid cells occupy the same
percentage of the boundary tissue in younger and
older men and there is a comparable total volume of
these cells per man (Table 1; Johnson et a!, 1986b).
The number of myoid cells per man tended to
decrease with age; however, significant differences
in number were not found by either of the methods
employed to calculate myoid cell numbers (Fig. 19).
Neither method indicated an increase in the number
of myoid cells. Likewise, the volume of a single myoid
cell is similar in younger and older men. Age-related
thickening of the tubular boundary tissue was not
due to an increase in boundary tissue itself (young
adult men had 3.5 0.2 ml and older adult men had
3.4 0.2 ml) but resulted from a disproportionate
reduction in tubular length (Fig. 10) compared with
the reduction in the number of myoid cells (Fig. 19).
The total volume of boundary tissue per man was not
increased with age (Johnson et a!, 1986b).
Sertoli cells are the somatic component of the semi-
niferous epithelium and are critical for the structural
support, nutrition and protection of germ cells (Dym
and M adhwa Raj, 1977; Feig et a!, 1980). W hile it is
generally accepted that the number of Sertoli cells is
stable in the adult (Clermont and Perey, 1957; Stein-
berger and Steinberger, 1971; Hochereau-de Reviers,
1981), there are seasonal fluctuations in the number
of Sertoli cells in stallions (Fig. 20; Johnson and
Thompson, 1983; Johnson and Nguyen, 1986; John-
No.6 SPERM ATOGENESIS AND AGING IN THE HUM AN- Johnson
347
Fig. 18. Electron micrographs of Leydig cells in a 66-yr-old man. These cells appear to be normal, with an abundance of smooth
endoplasmic reticulum (SER) and mitochondria (M). Also, the Golgi complex (GC) and lysosomal bodies (L) are present. Length of each bar
equals 2 m.
son, 1986) and an age-related decline in the Sertoli
cell population in humans (Table 1; Figs. 14 and 21;
Johnson et a!, 1984d). In stallions, there are more
Sertoli cells when daily sperm production is high;
there also is a slight but significant increase in the
ratio of germ cells to Serto!i cells in the breeding
season (Johnson and Thompson, 1983; Johnson,
1985b, 1986). In humans, the age-related decline in
D
1
PUW PER R0SS SEC11ON PERCENT P&CL
MC MC
n m
z
<1
w
Fig. 19. Effect of age on the number of
myoid cells, as calculated by two ap- ...J
proaches, in younger adult (20-48 yrs) ..J
and older adult (50-90 yrs) men. The LI-I
reduction in the number of myoid cells in 0
older men was not significant when based
on calculations including the number per -
cross section and tubular length. The 0
reduction in older men calculated from
the primary data of percent nuclei in the
parenchyma and the diameter (or volume)
of nuclei was significant (P <I 0.05). 0
(Johnson et al, 1986c).
LU
z
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NOV-JAN FEB-APR MAY-JUL AUG-OCT
the number of Sertoli cells is not associated with a
change in the ratio of germ cells to Sertoli cells (Table
1; Johnson et a!, 1984d). If the germ ce!l:Serto!i cell
ratio is an indication of the function of individual
Sertoli cells, these data may imply a seasonal increase
in Sertoli cell function in stallions but no decline in
the function of individual Sertoli cells in aging men.
Sertoli cells are evenly distributed in seminiferous
tubules throughout the human testis regardless of
age (Johnson et a!, 1984d). The average human Ser-
1.
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toli cell supports relatively few germ cells (Johnson et
a!., 1984d) compared with other species (Fig. 22;
W ing and Christensen, 1982; Johnson and Thomp-
son, 1983; Russell and Peterson, 1984; Johnson,
1986). Russell and Peterson (1984) found that the
elongated spermatid:Sertoli cell ratio averaged 6 to
14 for the rat, hamster, gerbil, guinea pig, rabbit, vole
and monkey; all had greater ratios than did humans
(Fig. 22). This paucity of germ cells (spermatocytes
and spermatids) accommodated by average individ-
ual Sertoli cells in humans is not reduced further
with advancing age (Table 1; Johnson et a!, 1984d). In
men 20 to 85 years of age, daily sperm production
was significantly correlated with the number of Ser-
toli cells per testis (Fig. 23). Ultrastructure of Sertoli
cells in aging men with active spermatogenesis (Fig.
24) is similar to that of younger men (Holstein and
Roose-Runge, 1981; Johnson et a!, 1984d) and to
males with 5a-reductase deficiency (Johnson et al,
1986a).
There are similarities between the stallion testis in
the nonbreeding season and the testis in aging men.
Both experience significant germ cell degeneration
during postprophase of meiosis (Johnson, 1985a;
Table 2) and both have significantly low numbers of
Leydig cells (Neaves eta!, 1984; Johnson and Thomp-
son, 1986) and Sertoli cells (Johnson, 1986; Figs. 14,
21). In conclusion, age-related changes in humans
include a reduction in the numbers of Leydig cells,
non-Leydig interstitial cells, myoid cells and Serto!i
cells. The decline in the number of Sertoli cells is
associated with a reduction in daily sperm produc-
tion.
A
AGE (years)
348 Journal of Andrology . November/December 1986 Vol. 7
Fig. 21. The relationship between number of Sertoli cells and
age in 71 men aged 20 to 85 years. (Johnson et al, 1984d).
Fig. 20. Number of Sertoli cells found
in 41 to 50 adult horses during each 3-
month period throughout one year. The
number of Sertoli cells per g was greater
(P< 0.05) from May to July than in other
periods. The number per testis was
greater from May to July than from
August to October or February to April
(P < 0.05) and than from November to
January (P< 0.01). (Johnson and Nguyen,
1986).
SPERM ATOGENESIS AND AGING IN THE HUM AN. Johnson
U
No. 6
Fig. 22. Ratio of the number of pri-
mary spermatids per Sertoli cell in the
rat, horse, and human. Data on rats was
taken from W ing and Christensen (1982),
horses from Johnson (1986), and humans
from Johnson et al (1984d).
E10L0
349
W flV4AT$
1
RAT HORSE HUMAN
Y = 18.8 + O.2X, rho = +0.70
Fig. 23. The relationship between the
number of Sertoli cells per man and daily
sperm production per man in 71 men
aged 20 to 85 years. (Johnson et al,
1984d).
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Possible Etiology of Age-related
Changes in Human Spermatogenesis
Sasano and Ichijo (1969) and Suoranta (1971) used
techniques of microangiography to illustrate the
relationship between age-related degeneration of
seminiferous tubular or Leydig cells and vascular
degeneration. Vascular degeneration was revealed
by microvascular lesions and diminished blood supply.
Regadera et al (1985) reported that arteriosclerosis
was involved in the decline of testicular function with
age. They found that elderly men with severe ath-
eromatosis had testicu!ar parenchyma that appeared
fibrosed with sc!erosed tubules. Hatakeyama et a!.
(1979) attributed some types of focal tubular atrophy
to locally limited ischemia. W e have had greater diffi-
culty obtaining good fixation by vascular perfusion of
testes from old men than from younger men obtained
from autopsy. However, it is not clear if vascular
#{246} 5#{212}0 idoo
NUMBER OF SERTOLI CELLS (mt I I tons)
Fig. 24. Electron micrograph of seminiferous epithelium in a 66-yr-old man. The Sertoli cells (SC) appear to be normal in the adluminal
compartment (Panel A) with a Golgi complex (CC) present in this view. Junctional complexes (JC) between adjacent Sertoli cells above the
basal compartment (Panel B), rod-like mitochondria (M), profiles of smooth endoplasmic reticulum (SER), rough endoplasmic reticulum
(RER), and lipid droplets (LD) of various densities characterize these Sertoli cells. Spermatogonia (SC) and Colgi spermatids with a
spherical nucleus called round spermatids (RS) are present between Sertoli cells. Bar length is equal to 2 m.
No. 6 SPERM ATOGENESIS AND AGING IN THE HUM AN- Johnson
351
insufficiency is the cause of tubular degeneration or
is itself a result of some other underlying process of
aging.
Given that inflammatory cells have been observed
in or around degenerating tubules in the testis of
aged men (Frick, 1969; Suoranta, 1971), that age-
related changes include focal mononuclear orchitis
(Honore, 1978), and that there is an increased per-
centage of older men with blood antibodies that
cross-react with spermatozoa (Hamer!ynck, 1970;
Fjal!brant, 1975), autoimmunity could cause tubular
degeneration (Harman, 1978). However, release of
antigens from already degenerated tubules could be
responsible for the autoimmunization (Harman,
1978). Age-related loss of Sertoli cells (Johnson et al,
1984d) could compromise the blood-testis barrier
and contribute to the release of sperm-specific anti-
gens.
The initial event of aging in the testis may take
place in the seminiferous tubules. Reduced activity of
Leydig cells has been demonstrated in several testicu-
lar disorders in which the seminiferous epithelium
was thought to be primarily involved (de Kretser et
a!., 1975). In case of a lack of advanced germ cells or
aplasia, the hCG response in young men is reduced
(Baker and Hudson, 1983). Tubule damage has been
associated with hyperresponsiveness of Leydig cells
to LH in rats (Jansz and Pomerantz, 1986). Progres-
sive deterioration of Sertoli cell function (Baker and
Hudson, 1983) or reduced numbers of Sertoli cells
(Figs. 12 and 20; Johnson et a!, 1984d) may cause
reduced daily sperm production. The decrease in the
number of primary spermatocytes or spermatids
occurs at the same rate as the loss in the number of
Sertoli cells with age (Johnson et a!, 1984d).
Reduced numbers of Sertoli cells could cause
reduced numbers of germ cells and reduced length of
tubules, which leads to increased thickness of the
tubular boundary tissue (Johnson et a!, 1986b).
Increased thickness of boundary tissue itself could
cause further damage by reducing the flux of metab-
olites or hormones entering, or waste products or
hormones leaving, the seminiferous epithelium.
However, the same underlying process could be
responsible for age-related losses of both Sertoli cells
and germ cells.
It is difficult to separate cause and effect in the
aging process. However, it is important that etiology
studies of the aging process be continued. Improving
our understanding of the aging process in the testis
should lead to a greater understanding of some forms
of infertility.
Conclusions
1. The efficiency of spermatogenesis (daily sperm
production per g parenchyma) is much lower in
humans than in other species.
2. Testicular parenchymal weight, percentage of
seminiferous epithelium, and dai!y sperm produc-
tion are significantly reduced in aging men.
3. Degeneration of germ cells occurs during sper-
matogonial mitosis, meiosis, and/or spermiogene-
sis in various species including humans.
4. Age-related reduction in sperm production occurs
during meiosis in humans.
5. Age-related reductions occur in human Leydig
cells, non-Leydig interstitial cells, myoid cells and
Sertoli cells.
6. The number of human Sertoli cells decreases with
age and is significantly correlated with daily
sperm production. However, there appears to be
no decline in function of individual Sertoli cells
present in older adult men as measured by the
germ cell:Sertoli cell ratio.
7. Vascular degeneration, autoimmunity, or loss of
Sertoli cells may contribute to the age-related
reduction of daily sperm production in humans.
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Seventh Annual M icrosurgery at the M ardi Gras
This symposium for Urologists, Andrologists, Pediatric Surgeons, and
Residents will be held at Southern Baptist Hospital, New Orleans, LA. It
will be in two parts. The first section, entitled M ICROSURGICAL LEC-
TURES AND LABORATORY, will be held February 26-28,1987, and will
emphasize a current series of M icrosurgical Applications in Urology-
Andrology. A 24-hour lab session will provide hands-on experience in
basic and advanced microsurgica! techniques.
The second lecture series, entitled CURRENT CONTROVERSIES IN
ANDROLOGY, will be held February 28, 1987, and include epydidyma!
function and epididymovasostomy, anti-sperm antibodies-fact or fic-
tion?, computerized semen analysis-is it clinically practical?, and more.
CM E credits offered through Southern Baptist Hospital.
For more information, planned lecture topics, and faculty, contact Beth
Norwood, M icrosurgery Lab Administrator, 1-800-621-1986, extension
75911; in Louisiana, 1-800-942-4330.