BIOLOGY OF REPRODUCTION 42, 55-62 (1990)

55
Effects of Chronic Ethanol Diet on
Pituitary-Testicular Function of the Rat1
uRIS SALONEN23 and ILPO HUHTANIEM I4
Departments of Anatomy3 and Physiology4
Institute of Biomedicine
University of Turku
SF-20520, Finland
ABSTRACT
W e studied the effects of a 6% ethanol liquid diet administered for 5 wk on the pituirary-gonadal function of
adult male rats. Because ethanol is known to reduce body weight, we used sucrose-fed animals as controls. No
significant differences in body, testis, or prostate weights were found between the rats exposed to ethanol and
their sucrose-ftd controls at the end of the 5-week treatment. Seminal vesicle weights decreased significantly
(p<O.O5) in the ethanol-treated group. Serum and testicular testosterone concentrations were sigmficantly re-
duced in the ethanol-treated group, to 43.6% and 48.3% of levels in the sucrose-fed controls, respectively
(p<O.O5). Serum luteinizing hormone (LII) and follicle-stimulating hormone (FSH) levels of the ethanol-treated
rats were 37.9% and 41.3% , respectively, of those of the sucrose-fed controls (p<O.OI - 0.05). The pituitary
levels of these hormones were similar to those of controls, but the ratios of pituitary LII and FSH to their serum
levels were clearly increased after ethanol exposure, to 492% and 206.1% , respectively (p<O.O5). In contrast,
pituitary prolactin (PRL) of the ethanol-treated rats was decreased to 40.2% (p<O.OI) of sucrose-fed controls.
Testicular content of LII receptors was significantly reduced (to 77.0% of controls; p<O.Ol), but content of FSH
receptors was slightly increased by the ethanol diet (to 121.5% of sucrose-fed controls; p<O.OS). No ethanol-
associated changes were apparent in testicular PRL and gonadotropin-releasing hormone (GnRH) receptors or
in pituitary GnRJI receptors. The increased ratio of pituitary/serum gonadotropins upon ethanol treatment indi-
cates that the release of gonadotropins is affected more than their synthesis. This effect is probably caused by
changes in hypothalamic GnRJI secretion, which may be sufficient to maintain the pituitary GnRH receptors and
gonadotropin synthesis, but not to sustain normal LH and FSH release. The present data on effects of a
5-wk ethanol exposure differ considerably from those found by us recently after a short-term exposure (7 days)
and e,nphasize that both in vivo models are important for studying effects of ethanol on male reproductive
functions.
INTRODUCTION
Ethanol intake is detrim ental to function of the pitu-
itary-gonadal axis in m ale rats (Boyden and Pam enter,
1983). A well-known consequence of chronic ethanol
treatment in males is that serum testosterone (T) usually
decreases. The cause of the lowered T m ay be intrates-
ticular, in part, i.e. direct suppression of testicular ster-
oidogenesis (Anderson et a!., 1983) or depletion of
testicular luteinizing horm one (LH) receptors and go-
nadotropin responsiveness (Gnanaprakasam et al., 1979;
Ganu et al., 1982). In addition, pituitary, hypothalam ic,
Accepted August 18, 1989.
Received October 27, 1988.
tThis study was supponed by a grant from The Rcseanth and Science
Foundation of Farmos, The Academy of Finland, and The Sigrid Juselius
Foundation.
2Rep .i t requests: lins Salonen. Dept. of Anatomy. Inst. of Biomedicine,
Univ. of Turku. Kiinamyllynkatu 10. SF-20520 Turku, Finland.
and m ore central defects have been proposed as primary
causes of the lowered activity of the pituitary-testicular
axis (Bannister and Lowosky, 1987). Indeed, Dees et a!.
(1983, 1984) found that after i.p. ethanol treatment for
8 days, the hypothalamic gonadotropin-releasing hor-
m one (GnRH) content was increased. W hen this in-
crease is combined with the decreased LH as a result of
the sam e or shorter ethanol exposure (Dees et al., 1983;
Cicero et al., 1979), a disturbance in the hypothalamic
GnRH release is likely.
Our recent study on short-term (7 days) effects of
ethanol revealed a num ber of changes in the hypotha-
lam ic pituitary-gonadal function in m ale rats and sug-
gested that ethanol does not hamper hypothalam ic
GnRH release and thereby decrease levels of LH and T
(Salonen and Huhtaniemi, 1988). Instead, the previous
study indicated that decreased testicular LH and prolac-
tin (PRL) receptor levels m ay be responsible for the
inadequate T secretion. W e wanted to test this theory in
56
SALONEN AND HIJHTANIEM I
the long term , since a short-term experiment may be
com plicated by stress superimposed on the effects of
ethanol. Also, adaptation of the endocrine functions to
chronic ethanol intake m ay m odulate long-term ethanol
effects. To investigate the specificity of the eventual
receptor-depleting effect of chronic ethanol exposure in
the testis, we determ ined testicular follicle-stim ulating
horm one (FSH) and GnRH receptor levels. Further-
m ore, we carefully controlled the weight loss associated
with ethanol exposure by using sucrose-fed control
anim als in this experiment
M ATERIALS AND M ETHODS
A total of 28 m ale W istar rats, 12 wk old, were
studied. They were housed under constant conditions of
light (on at 0600 h, off at 2000 h) and tem perature
(21 - 22C). The rats were caged in pairs and divided
into three groups: control, ethanol-treated and sucrose-
fed control groups. The original weights of these ani-
m als were 306 12.9 (m ean SEM ), 283 11.1 and
293 7.6 g, respectively.
The control group (n=10) was allowed standard labo-
ratory chow (Oy Hankkija AB, Koippi, Finland) ad
libitum . The ethanol-treated group (n=8) received the
ethanol diet where pure ethanol was diluted into RN
liquid nutrim ent (Sem per Ab, Stockholm , Sweden) to
make a 6% (v/v) solution (100 ml liquid nutrim ent + 13
ml ethanol + 103.5 m l distilled water). The RN nutri-
m ent contained 50 g/l protein, 40 g/l fat, and 160 g/l
carbohydrates. Ethanol thus constituted 35.6% of total
calories. The sucrose-fed controls (n=10) received iso-
caloric diets in which sucrose replaced ethanol. Each
pair of animals received the sam e volum e (per original
weight of anim al) as each corresponding experim ental
pair had eaten on the previous day (hence, pair-fed
controls). The liquid nutrim ent was fed in regular water
bottles. In addition, water was allowed ad libitum to all
animals. Rats were adapted to the liquid diets during 1
wk, and the experim ent was carried out .during the
following 5 wk.
Blood sam ples (50 jil capillary blood diluted into
1000 I.tl distilled water) for determ ining ethanol concen-
tration were taken under sodium pentobarbital anesthe-
sia from the orbital plexus of the ethanol-exposed rats
at 0400 h (n=4) and 2200 h (n=5) at the end of the
fourth week of the diet The animals from the sucrose-
fed control group were anesthetized correspondingly.
However, blood was not drawn from these rats, since
this difference in stress was not expected to affect the
hormone status at the time of exsanguination, after 1
wk. Ethanol content was analyzed by head-space gas
chrom atograph (Eriksson et a!., 1977).
On Day 34 at 1500 h, ethanol adm inistration was
discontinued, and the ethanol-treated rats were given
the sucrose liquid diet. Rats were killed by decapitation
on Day 35 at approxim ately 1100 h. Trunk blood was
collected, and serum was separated by centrifugation
and stored at - 20C until required. The right testis,
ventral prostate, and sem inal vesicles were excised and
weighed, and the left testis was snap-frozen in liquid
nitrogen and stored at - 70C.
Hormones and Reagents
Highly purified human choriom c gonadotropin
(hCG; CR-121; 13,500 lU/m g by bioassay) was pre-
pared by Dr. R. Canfleld (Colum bia University, New
York, NY) and supplied by the Center for Population
Research, NICHHD (Bethesda, M D). Partially purified
hCG (Pregnyl; 3000 Hi/mg) was purchased from Orga-
non (Oss, The Netherlands). Highly purified hum an
FSH (hFSH; corresponding to 7000 IU of W HO 69/104
per m g by bioassay) was donated by Dr. P. A. Torjesen
(Aker Hospital, Oslo, Norway). A partially purified
preparation of hum an FSH (M etrodin; 150 lU/m g) was
donated by Ares-Serono (Rom e, Italy). Purified hum an
growth horm one (hGH, NIADDK-I-1) was donated by
the National Horm one and Pituitary Program and
NIADDK (Bethesda, M D). Partially purified ovine PRL
(oPRL; 30 lU/m g) was purchased from Sigm a Chem i-
cal Com pany (St Louis, M O). Testosterone antiserum
was a gift from Prof. R. Vihko (University of Oulu,
Finland). Tritiated T [NET-5-53; 1,2,6,7,16,1 7-3H(N)]
was purchased from New England Nuclear (Dreichen,
F.R.G.). The GnRH agonist analog (buserelin; D-Ser-
(tBu)6-des-GlyO GnR}1 N-ethylam ide; GnRH-A) was
donated by Hoechst AG (Frankfurt am M ain, F.R.G.).
Hormone M easurements
Serum T was m easured after diethyl ether extraction
by radioim m unoassay (RIA), as described by Ham -
m ond et al. (1977). Cross-reactivity of dihydrotestoster-
one (DHT) with the T antiserum was 64% , and it was
not corrected for (J#{228}nne et a!., 1974). Testicular T was
measured by the sam e RIA m ethod from the supem a-
tants of the testis hom ogenates (obtained at 25,000 x g,
see below). Serum and pituitary LH, FSH, and PRL
were determined by double antibody RIAs, using the
NIADDK assay kits and standards, as described before
(Clayton and Bailey, 1982). The horm one levels were
expressed in terms of the NIADDK-RP-2 standard
preparations.
PITUITARY-TESTICULAR EFFECFS OF ETHANOL 57
M easurement of Testicular
FSH, LII, and PRL Receptors
Highly purified hFSH, hCG, and hGH were radioio-
dinated by a solid-phase lactoperoxidase method, as
described by Karonen et al. (1975), and purified on a
0.5 x 10-cm column of Bio-Gel P-60 (BioRad, Rich-
m ond, CA). Specific activity and proportion of radioac-
tivity associated with biologically active horm one were
assessed as described by Cart et a!. (1976). The decap-
sulated testes were homogenized in 10 ml/g tissue of
Dulbeccos phosphate-buffered saline (pH 7.4) plus
0.1% bovine serum albumin (PBS/BSA), and the homo-
genates were used as such for FSH, LH, and PRL
receptor measurements. One-hundred microliter au-
quota of homogenate were equilibrated (overnight at
23C) with 60,000 cpm (about 1 ng) of [ I]iodo-hCG,
100,000 cpm (about 3 ng) of [ I]iodo-FSH, or 60,000
cpm (about 2 ng) of [ IJiodo-hGH. For assessing
nonspecific binding of hCG, hFSH, and hGH, m atched
sam ples were incubated in the presence of 50 IU Preg-
nyl, 1.5 IU M etrodin, or 5 ig oPRL, respectively.
Bound and free hormone were separated, after
15-fold dilution of the sam ples with PBS/BSA, by
centrifugation (Huhtaniemi et al., 1984). The use of
hGH to measure PRL receptors was based on the ability
of hGH to bind specifically to receptors of PRL, but not
to those of GH, in rats (Aragona and Friesen, 1975;
Posner, 1976). Since Scatchard analysis of LH, FSH,
and PRL receptors revealed similar binding affinities in
different experimental groups (results not shown), the
single point measurements correlate to, although are not
identical with, the number of binding sites in the tissue
sam ples.
M easurement of Testicular and
Pituitary GnRR Receptors
GnRH receptors were measured as described before
(Clayton, 1983; Huhtaniemi et al., 1987). In short,
100-p.! aliquots of a partially purified m em brane prepa-
ration (25,000 x g pellet of a 600 x g supernatant,
prepared from testicular hom ogenates) were equili-
brated with 65,000 cpm of radioiodinated GnRH-A at
4C for 90 m m . The pituitary receptors were assayed
from crude tissue hom ogenates, with 95,000 cpm of
[ I]iodo GnRH-A/tube.
Statistical Analysis
The data were analyzed statistically by analysis of
variance (BM DP 7D; BM DP Statistical Software, Uni-
versity of California). W hen variances of the groups
were not equal, W elch and Brown-Forsythe tests were
used (ANOVA; BM DP 7D). W hen differences between
groups were found, the significance of these was ana-
lyzed by the t-testof the 7D program.
RESULTS
Ethanol content of blood from rats on the 6% ethanol
diet was 31.0 7.4 mmol/l (n=5) at 2200 h and 47.5
7.6 (n=4) at 0400 h at the end of the fourth week of the
diet
There were no significantdifferences in body, testis,
or prostate weights between the ethanol-treated animals
and the sucrose-fed controls (Table 1). However, the
weight of the seminal vesicles was significantly lower
in the ethanol-treated rats than in either the sucrose-fed
or untreated control rats (p<O.O5 and p<0.01, respec-
tively).The body weights of the untreated controls were
significantlyhigher than those of the other two groups,
(p<O.OS and p<0.00l, respectively).
Serum and testicularT concentrations in the ethanol-
treated group were significantly (p<O.OS) lower than
those in the sucrose-fed group (43.6 and 48.3% of the
respective control values) (Fig. 1). Serum LH and FSH
of the animals receiving the 6% ethanol diet were
likewise significantly decreased com pared to the values
TABLE 1. Body weights and weights of testes, ventral prostates. and seminal vesicles in male rats after 5 wk of 6% ethanol diet (mean SEM ).
Diet n
Final weight
(% of original weight)
Testis
(g)
weight Ventral prostate
weight (g)
Seminal vesicle
weight (g)
Control 10 129.0 2.8k 1.71 0.06 0.36 0.03 0.48 0 03b
6% ethanol 8 103.2 13.0 1.55 0.18 0.27 0.04 0.37 0.04
Sucrose 10 110.7 1.4 1.63 0.03 0.32 0.03 0.46 0.02
F 15.51 1.73 2.36 4.71
Significant difference (p<O.05) with sucrose-fed group.
bsigni&ant difference (p. O.0l) with 6% ethanol-treated group.
cSignjflCant difference (pd).0OI) with sucrose-fed group.
F= M ean square bet en classe mean square within classes.
Control Diet Ethanol
58
SALONEN AND HUHTANIEM I
a)
C
0
L.
a)
0
wo
a)
(1 )
G)UI
It 0)
4-.
(l)
0
4-.
U .,
FIG. 1. Serum (upper panel; F=431, ANOVA) and testicular (lower panel.
F=3.63) testosterone conceniratious (mean 5EM ) in control rats (n=10), an-
crose-fed rats (Diet. N=10). and 6% ethanol-treated rats (Ethanol; n=8) after 5
wk. M tertskr indicate significant differences between the ethanol-treated and
sucrose-fed control groups: pd).O5; p<O.Ol: ***p<O.001).
of the sucrose-fed animals, down to 37.9% (p<0.01)
and 41.3% (p<0.05), respectively (Fig. 2). In contrast,
although not statistically different, the pituitary FSH
and LH contents of the ethanol-treated rats were
179.1% and 114.8% of those in sucrose-fed controls,
and the proportions of pituitary LH and FSH to their
serum levels were significantly increased over those of
the sucrose-fed controls (to 492% and 206.1% , respec-
tively, p.<0.05) after ethanol exposure. There were no
significant changes in serum PRL levels, but pituitary
PRL in the ethanol-treated rats was decreased to 40.2%
(p<O.Ol) of the sucrose-fed controls.
The testicular LH receptors of the ethanol-treated
rats were significantlydecreased, to 77.0% of those of
sucrose-fed controls (p<O.Ol; Fig. 3). Testicular FSH
binding was found to be increased (to 1213% of su-
crose-fed controls, p<0.05), but the PRL receptors did
not differ from those of the sucrose-fed controls (Fig.
3). No ethanol-associated changes were found in testic-
ular or pituitary GnRH-A receptors of ethanol-treated
rats compared to the sucrose-fed controls (Fig. 4).
In sucrose-fed rats, serum FSH was significantly
increased, and the ratio of pituitary to serum FSH was
decreased (p<0.05 and p<0.01, respectively) from levels
in ad libitum -fed controls. Testicular LH receptors
(p<O.OO1) and pituitary GnRH receptors (pCZO.OS) were
significantlydecreased in the sucrose-fed diet rats ver-
sus the values for the ad libitum -fed control rats.
DISCUSSION
The pituitary GnRH receptor content was not
changed by the 5-wk ethanol diet, which is in keeping
with previous findings after short-term ethanol expo-
sure of m ale (7 days; Salonen and Huhtaniemi, 1988)
and fem ale rats (4 days; Retton et a!., 1987). The
apparent decrease in these binding sites versus controls
is due solely to food restriction, as has been docu-
m ented previously (Salonen and Huhtaniemi, 1988;
Bergendahi et a!., 1989). In contrast, serum LH and
FSH levels were decreased in the ethanol-treated rats
m ore distinctively than after a previous ethanol treat-
m ent for 7 days (Sa!onen and Huhtaniem i, 1988). These
data emphasize the necessity of pair-fed control ani-
mals. Had the ethanol-treated animals been com pared
with animals fed ad libitum , a biased picture of the
specific effects of ethanol would have em erged.
Since the reduced serum LH cannot be due to en-
hanced negative long-loop feedback of T (serum T was
reduced), the defect m ost likely resides in the GnRH
release or its effects on pituitary function. The unaltered
pituitary GnRH receptor levels and the close correlation
between hypothalamic GnRH release and its receptor
numbers (Clayton and Can, 1981) suggest that the
maintaining effect of GnRH on its receptor levels was
not ham pered. However, it is possible that the pul-
satility of GnRH release is altered in such a fashion that
it is unable to support norm a! gonadotropin release.
Therefore, our results on long-term effects of ethanol
do not support total loss of GnRH release, as has been
suggested by Dees et a!. (1983, 1984). This discrepancy
is probably due prim arily to the fact that these authors
did not use diet controls. Besides changes in GnRH, the
nature of gonadotropins synthesized and secreted could
be directly or indirectly affected by ethanol. This al-
tered microheterogeneity could then alter the clearance
of the circulating gonadotropins. However, increased
I+.0
2.0
I
20
15
10-
5.
0-
I H
Control Diet Ethanol
Control Diet Ethanol
FIG. 2. Serum (kftpwie() LII (F=4.86. ANOVA). FS}I (F=5.86), and PRL (F=0.58) (mean SEM ) and pituitary (rig& panel) LII (F=2.42). FSH (F=1 .02), and
PRL (F=9.84) in control rats (N=8 - 10), sucrose-fed rats (N=9 - 10) and ethanol-exposed rats (N=8). Asterisks above the bars indicate significant differences in
serum gonadouopins between ethanol-treated and sucrose-fed groups. Significant increases (LII [F=4.94) and FSH (F=5.73J) or decreases (PRL; F=3.07) in the
ratios of pituitary to serum gonadotropins are indicated by asterisks below the figure. (Symbols defined in legend to Fig. 1.)
PITUrrARY-TESTICULAR EFFECTS OF ETHANOL
clearance of FSH and LH from the blood does not
provide a satisfactory explanation to their pituitary
changes and to the increased pituitary/serum ratios of
I
U)
U-
E
L.
a)
U)
0
-J
a-
E
J
U)
(I)
59
LH and FSH. These suggest that release of gonadotro-
pins from the pituitary is impaired, which is probably
related to deficient GnRH stimulation.
>
L
0
4-.
0.
I
(I)
Li-
>
L.
0
4-.
4-.
a.
-J
a-
A m ore long-term ethanol diet than the present (5%
ethanol for 164 days) previously resulted in increased
plasma FSH compared to plasma FSH in isocalonc
control rats (Van Thiel et al., 1979; Gavaler et al.,
1980). However, FSH levels were not different in
weight-starved controls (i.e. controls fed a standard
chow at a rate such that the weight paralleled that of the
alcohol-fed animals), and plasma LH was unchanged.
U)
0
a
ci)
0
a)
a:
I
-J
Also, lower (Cicero and Badger, 1977; a 20-day diet
with increasing ethanol content) and higher (Bannister
and Lowosky, 1987; hum an alcoholics) plasm a LH
values have been reported. These exam ples indicate that
changes in gonadotropin levels depend on diet, type of
controls, length of ethanol exposure, and species.
The decreased pituitary content of PRL after adm in-
istration of the 6% ethanol diet for 5 wk contrasts with
changes in pituitary LH and FSH. It is presum ably due
to the deleterious effects of ethanol on the metabolism
and receptors of dopam ine (Reggiani et a!., 1980), a
major inhibitory m odulator of PRL secretion (Ben-
Jonathan, 1985). This finding also indicates, as with
gonadotropins, that ethanol affects the hypothalamic
control of PRL release, which is m ostly negative.
In contrast to changes of the ligand horm ones, there
was a dissociation between the responses of testicular
LH and FSH receptors. LH receptors decreased, as has
been shown with alcohol exposure (Gantt et a!., 1982;
Salonen and Huhianiemi, 1988). In rats, suppressed
12:
040
0
L.
C
0
0
0
120
80
ci)
40
Controt Diet Ethanol ControL Diet Ethanol
60
SALONEN AND HUHTANIEMI
U )
0
a-
ci)
0
ci)
a:
I
c-n
LL
100
o60
0
C
0
0
4.-
0
0 #{149}
20
0
FIG. 3. Contents of receptors for LII (F=35.12, ANOVA), FSH (F=3.79),
and PRL (F=1.34) (mean SEM ) in testes of control rats (N=10), sucrose-fed
rats (N=I0), and ethneol-trea&ed rats (N=7 -8). The receptor levels were calcu-
lated per testis, and the mean of the controls was taken as 100% . (Symbols
defined in legend toFIg. 1.)
U)
0
a
ci)
C-)
ci
a:
I
a:
C
0
0
(n-.1
- 6
ot-
- -
a
80
1040
CD
FIG. 4. Contents of testicular (upper panel) and pituitary (lower panel)
receptors for GnRH (mean SEM F=0.98 and 4.77. respectively, ANOVA) in
control (N=l0). sucrose-fed (N=9 -10). and ethanol-treated (N=8) rats. (Sym-
b o L t defined in legend to Fig. 1; data are expressed as in FIg. 3.)
PITUITARY-TESTICULAR EFFECTS OF ETHANOL
61
circulating PRL, which results in depletion of LH re-
ceptors (Aragona et al., 1977; Huhtaniemi and Catt,
1981), could not be dem onstrated as the cause of the
LH receptor loss in the present experiment. In a m ore
chronic situation (as in the present experiments), factors
other than PRL - probably LH itself - m ay contribute to
maintenance of the LH receptors. However, direct go-
nadal effects of ethanol cannot be excluded as the cause
of this receptor change.
Although testicular FSH receptor levels increased
during the experim ent, they had not changed during the
7-day ethanol treatment (Salonen and Huhtaniemi,
1988). The mechanism of this finding remains obscure
since our knowledge, in general, about the regulation of
testicular FSH receptors is very limited. It is tem pting
to speculate that this phenom enon represents a testicu-
lar compensatory mechanism for decreased FSH secre-
tion. The suppressive effect of a long-term ethanol diet
on testicular LH receptors is thus selective, since it did
not reduce testicular FSH, PRL, or GnRH receptors.
Previous studies have indicated that the endocrino-
logic effects of acute ethanol treatment differ from
those of chronic administration (Bannister and Lowo-
sky, 1987). In the current study, the effects of ethanol,
especially on serum and testicular T and on serum
gonadotropins, were uncompensated and more pro-
nounced than those found after exposure for 7 days
(Salonen and Huhtanierrn, 1988). On the other hand, the
effects of 7-day ethanol treatment on testicular PRL and
GnRH receptors were different from the present results,
probably reflecting endocrine adaptation of the animals
to a longer exposure to ethanol.
Additionally, diet restriction itself appeared to de-
crease the testicular LH and pituitary GnRH receptor
levels, in accordance with previous findings (Salonen
and Huhtaniemi, 1988; Bergendahl et al., 1989). The
levels of serum T, LH, and FSH in the sucrose-fed
anim als, although not significantly elevated com pared
to controls, were different from horm one levels after
short-term (usually 1 wk) starvation (Howland and
Skinner, 1973; Pirke and Spyra, 1982). However, stud-
ies on the endocrine effects of m ild food restriction for
5 wk, as that in sucrose-fed controls, are not available.
The tendency of mean levels of serum T, LH and FSH
to increase in the sucrose-fed controls contributes to the
changes observed in the ethanol-treated group. Howev-
er, this confounding does not concern the m ost impor-
tant parameter, ethanol-induced suppression of testicu-
lar T secretion,and does not change the conclusions on
the effects of ethanol.
Acute withdrawal of ethanol (18 h before killing)
could constitute a stress factor that affects hormone
levels, and stress and increased corticosteroid levels are
known to reduce serum T (for references, see V#{228}limaki
et al., 1984). Indeed, in m ice, ethanol-withdrawal sym p-
tom s and increased corticosterone concentrations have
been reported 6 h after the rem oval of ethanol from the
diet; however, corticosteroid levels returned to control
level by 24 h after ethanol withdrawal (Tabakoff et at.,
1978) and thus m ay not be of significance for the
present experiment. Similar increases are associated
with ethanol consum ption (Van Thiel, 1980; Boyden
and Pamenter, 1983); therefore, a m odel in which the
animals are maintained on ethanol until decapitation is
not a better alternative.
In summary, the present results support the view that
a chronic ethanol diet decreases pituitary LH and FSH
release, probably by subtle m echanism s m odifying hy-
pothalam ic GnRH release to the extent of decreasing
gonadotropin release, but does not suppress their syn-
thesis and m aintenance of GnRH receptors. At the
testicularlevel, LH receptors are a particularly vulnera-
ble site of ethanol action. The fact that ethanol acts at
several sites is reflected by the discordant responses of
reproductive organ weights (Table 1; Figs. I and 2).
However, the values of testicular T and weights of
seminal vesicles are related. The low serum LH and
reduced testicular LH receptors explain the low serum
and testicular T, but do not exclude possible concom i-
tant direct effects of ethanol on testicular steroidogene-
sis.
ACKNOW LEDGMENTS
The authors are grateful to M s. Peppi Sievers. M s. Aila M ets#{228}vuori.M s.
Raija Andersen. M s. Pirkkokauham5ki, and M s. Tarja Saari for technical assis-
tance.
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