Inhibitory effect of chlorpromazine on rat reproduction: A test of administration for nine weeks before breeding

Reproductive Toxicology, Vol. 4, pp. 29-36, 1990

0890-6238/90 $3.00 + .00 Copyright © 1990 Pergamon Press pie

Printed in the U.S.A.

INHIBITORY EFFECT OF CHLORPROMAZINE ON RAT REPRODUCTION: A TEST OF ADMINISTRATION FOR NINE WEEKS BEFORE BREEDING KEIYA SAIGO Department of Pharmacology, School of Dentistry, Iwate Medical University, 3-27, 1 Chome, Chuodori, Morioka 020, Japan Abstract -- The effect of chlorpromazine (CPZ) administration prior to insemination on reproduction was studied using male and female Sprague--Dawley strain rats evaluated over three generations. CPZ in doses of 12.5, 25, 50, and 100 mg/kg was administered to both male and female rats orally every day for 9 weeks before mating with the following results: 1) The duration of the estrous cycle was prolonged in the F o females. 2) The incidence of insemination and pregnancy of the F o males and females as well as the number of surviving fetuses and newborns showed a tendency to decrease. 3) The body weight of the rats in Fo and F~ generations decreased depending on the dose of CPZ, but the body weight of the F 2 generation increased. 4) The wet weight of the major organs of the rats in F o and F 1 generations was affected by administration of CPZ at 100 mg/kg. The weights of the liver, kidney, adrenal, pituitary, testis, and prostate glands in males and the kidney, adrenal gland, and ovary in females were increased, whereas the weights of the pituitary and the uterus were decreased in females. These results suggest that premating administration of CPZ to the parent rats exerts influence on the reproduction of the F o and F~ generations. Key Words: Chlorpromazine;reproduction; prematingadministration; rat.

of 22 to 26°C, a humidity of 50% to 65%, and a 12-h light-dark cycle with the light period from 7:00 to 19:00 h. Solid feed for maintenance and for reproduction (Japan Clea Co.) and tap water were supplied to the rats ad libitum. Chlorpromazine hydrochloride (Shionogi Seiyaku Co., Japan) was used as the test drug. The CPZ was dissolved in distilled water, preserved in a brown bottle in a cool dark place and prepared every two days. The test doses of CPZ were 12.5, 25, 50, and 100 mg/ kg/day, given in a volume of 0.5 mL/100 g of body weight through a metal needle used for gavage. CPZ was administered intragastrically once daily. In the control group, the same amount of distilled water was administered using the same schedule and technique.

INTRODUCTION

Chlorpromazine (CPZ) was introduced to psychiatric therapy about 40 years ago and has played a central role in psychotropic drug therapy in subsequent years (1). Despite the use of this drug for many years as a therapeutic agent, the influence of CPZ on reproduction has not been studied extensively except for tests of fetal toxicity (2-5). The influence of CPZ exposure on subsequent generations has not been reported except for the studies of Itoh et al. (6, 7) on the effect of CPZ on rat reproduction and of Fujii (8) on the functional effects of maternal exposure to CPZ on the first and second generations in rats. In the present study, the effect of premating administration of CPZ on male and female rats' subsequent reproductive capacity was examined.

Procedures

MATERIALS AND METHODS

Groups of 10 male and 25 female rats at 6 weeks of age (Fo generation) were maintained separately. From 10 weeks of age, CPZ was administered for a 9-week period. During the experimental period, the general condition of the rats was observed daily, and body weight as well as food and water intakes were recorded every week. A smear test for evaluating the estrous cycle was undertaken for 2 weeks before CPZ administration, and between the 4th and 5th weeks and between the 8th and 9th weeks after CPZ administration, respectively. Mating was accomplished by housing 1 male with 2 or 3 females in a single cage for 2 weeks from the day after

A n i m a l s and drug

Male and female Sprague Dawley strain rats, purchased from Japan Clea Co. were used in the experiments. The rats were housed in a habitat at a temperature

Address correspondenceto: Keiya Saigo, Departmentof Pharmacology, School of Dentistry, Iwate Medical University, 3-27, 1 Chome, Chuodori Morioka 020, Japan. Received 6 March 1989;Revision received 7 June 1989;Accepted 20 June 1989. 29

30

Reproductive Toxicology Table 1. Insemination and pregnancy data for Fo rats exposed to chlorpromazine for 9 weeks before breeding Chlorpromazine, mg/kg/day

Observation N of rats (Fo) Male Female Insemination (%)

0

12.5

25

10 10 10 17 19 17 100.0 100.0 94.1

50

100

10 21 85.7

10 25 84.0

N of female rats inseminated/N of female rats paired with male (17/17) (19/19) (16/17) (18/21) (21/25) Pregnancy (%) 94.1 94.7 81.3 72.2 57.1" N of pregnant rats/ N of female rats inseminated

Volume 4, Number 1, 1990 The remaining males and females were maintained separately. At the age of 10 weeks, 5 males and 10 or 14 females of different litters per group were mated. The F 2 rats were thus obtained and studied in the same fashion as the F 1 generation. The teratogenicity of CPZ was not evaluated in the present study.

Stat&tical analysis The significance of differences between the treated and control groups was evaluated using Student's t test, the multiple comparison analysis of Kruskal-Wallis, and the direct probability analysis of Fisher. RESULTS

(16/17) (18/19) (13/16) (13/18) (12/21)

*Significantly different from 0 mg/kggroup, p < 0.05.

the last dose of CPZ. Mating was allowed between males and females that had been administered the same dose of CPZ. Insemination was confirmed by detection of spermatozoa in the vaginal smear test, and the day of detection was defined as "Day 0 " of pregnancy. One third of the pregnant rats were sacrificed by chloroform anesthesia on the 14th day of pregnancy and one third were sacrificed on the 21st day of pregnancy and underwent immediately autopsy. The number of live fetuses, the number of dead fetuses, and the number of myometrial glands were recorded, and the gross morphology of the uterus and ovary was studied. At the same time, the heart, lung, liver, kidney, spleen, adrenal gland, whole brain, pituitary gland, and submaxillary gland were removed and evaluated by wet weight, appearance, and gross morphology. Male rats underwent autopsy in a manner similar to female rats with additional gross observations of the testis and prostate gland. The remaining one third of the pregnant rats were allowed to deliver litters (F 1 generation) in a natural course. Before, during, and immediately after delivery, the mother rats were protected from outside stimuli as completely as possible with careful avoidance of environmental changes and other stimulation. The mode of rearing the pups and the growth of the pups were observed. In a similar study method, Park (9) adjusted the litter size to six members. In the present study, the litter size of the F~ pups was limited to eight members with four males and four females. The selection of litter members was made in a random fashion at 48 h after birth. A similar litter standardization procedure was repeated for the litter size of F 2 pups. After weaning of the F 1 rats, all Fo mothers and fathers and ten of the F1 rats (males and females, respectively) underwent autopsy using the same method as previously described.

F o generation Estrous cycle. The estrous cycle ranged from 4 to 10 days prior to CPZ administration, 4 to 10 days after the administration of 12.5 mg/kg, 4 to 11 days after administration of 25 mg/kg, 5 to 13 days after the administration of 50 mg/kg, and 5 to 13 days after the administration of 100 mg/kg. Thus, the estrous cycle was prolonged with CPZ administration in a dosedependent fashion. Incidence of insemination and pregnancy. As shown in Table 1, the incidence of insemination and pregnancy showed a tendency to decrease with the CPZ dose in a dose-dependent fashion. The incidence was especially lower in the group of mating rats given 100 mg/kg of CPZ, with a pregnancy rate of 57.1%. Autopsy findings on the 14th and 21st days of pregnancy. The results of the autopsy performed on the 14th day of pregnancy are shown in Table 2. The number of myometrial glands per litter in the groups of rats receiving CPZ was not different from the control group, but the weight of the live fetuses decreased in a doserelated pattern. The number of live fetuses significantly decreased in the mating group of rats given 100 mg/kg of CPZ. The results of the autopsy on the 21st day of pregnancy are also shown in Table 2. The number of myometrial glands per litter in the treated group was not different from that in the control group. The number of live fetuses significantly decreased in mating groups of rats given 12.5, 25, and 100 mg/kg of CPZ. The weight of the live fetuses significantly increased over the control group in mating groups of rats given 12.5 or 25 mg/kg of CPZ, but significantly decreased in the mating groups of rats given 100 mg/kg of CPZ. The uterus and ovaries showed no visible external abnormalities in gross observations.

General condition. During CPZ administration,

Breeding effect of chlorpromazine• K. SAIC,O

31

Table 2. Autopsy findings on day 14 and 21 of pregnancy in Fo rats exposed to chlorpromazine for 9 weeks before breeding Chlorpromazine, mg/kg/day (Mean ± SE) Day

Observation

0

14th

N of dams (Fo) Myometrial glands/ litter Live fetuses/litter

6 12.7 --- 1.4 12.2 ± 1.2 2.68 ±0.02 6 13.0 ---0.8 12.7 ±0.2 5.03 ---0.03

Average live fetal weight (g) 21st

N of dams (Fo) Myometrial glands/ litter Live fetuses/litter Average live fetal weight (g)

12.5

25

50

100

6 12.0 -+0.8 11.5 ---0.8 2.65 ---0.03

4 13.6 +- 1.0 13.2 ± 1.2 2.48* -+0.02

4 12.8 +0.9 12.0 ± 1.0 2.39** ---0.05

4 10.3 ± 1.0 9.7* ---0.8 2.12"* ±0.04

6 11.8 +0.8 11.7" ±0.1 5.34** ±0.04

5 11.2 - 1.0 10.8" ---0.1 5.21" ___0.05

4 12.8 ---0.2 12.2 ±0.4 4.94 ±0.17

4 10.2 ±0.6 9.3* ± 1.3 3.84* ±0.26

* and **: significantly different from the 0 mg/kg group, p < 0.05 and p < 0.01, respectively.

spontaneous locomotion was inhibited, and a sedated state was noted about 5 to 10 min after administration of CPZ to the male and female groups. These symptoms persisted for about 30 min to several hours, with the intensity and the duration depending on the dose and occurred with every CPZ administration. After the 2nd week of administration, however, the symptoms were somewhat alleviated and the duration of symptoms following each administration slightly shortened. In male and female rats given 100 mg/kg CPZ, a blepharitis appeared in about 25% of rats about 4 to 6 weeks after the beginning of CPZ administration. The manifestation of the blepharitis were more severe in males than in females. This symptom persisted throughout the period

Fig. 1. Blepharitis developed at left side about 5 weeks after chlorpromazine (100 mg/kg) administration to male rat.

of CPZ administration without subsiding (Figure 1).

Body weight, and food and water intake. Figure 2 shows the course of changes of body weight in each male and female group of rats. In both male and female groups, the increase in body weight was inhibited depending on the dose of CPZ. The inhibition was especially pronounced in male rats receiving 100 mg/kg CPZ. Food and water ingestion decreased depending on the dose of CPZ in a pattern similar to body weight. Flgeneration Growth of the F 1 rats. The F 1 rats from the F o parents exposed to CPZ were studied. The length of pregnancy of the F o mothers was 22.0 to 23.5 days without any difference with varying doses of CPZ as shown in Table 3. The number of live pups (F 1 generation) per litter was significantly lower in the F o rats given 100 mg/kg CPZ than in controls. The body weight of the F 1 pups 48 h after birth was significantly decreased compared to controls, in a dose-dependent fashion as shown in Table 3. This tendency was again noted in the subsequent increase in body weight (Fig. 3). The survival of the F 1 pups on the 22nd day after birth is shown in Table 3. A significant decrease in survival compared to the control group was noted in the F 1 pups resulting from mating F o rats given more than 50 mg/kg of CPZ. Most of the deaths of the F x rats occurred within several days after birth, and none occurred later than 2 weeks after birth. Growth of the F~ rats after weaning was not different from that of the control group. The average body weight of the F~ rats at about 10 weeks of age showed no difference from that of the control

32

Reproductive Toxicology

I

Volume 4, Number 1, 1990

male

40

40[

female

30

.~ 20

"~ 20

a oD e-

._~

10 0

m 5

6

7

8 9 Weeks

I

1

2

3

4

5

6

7

8

-10' [

9

Weeks

-10

~ Chlorpromezlne 0 0--0 12.5 26 50

-20

100 mg/ko

0'.--(3

-30

rng/kg mg/kg mg/kg nl~/kg

Fig. 2. Rates of weight gain of the male and female rats receiving chlorpromazine for 9 weeks before breeding.

group. During the period of observation, no abnormal behavior was noted in the F 1 rats. The estrous cycle at the age of 10 weeks was 4 to 10 days with no difference from the control group. Gross observations of the F1 rats revealed no visible external abnormalities.

Incidence of insemination and pregnancy. As shown in Table 4, the incidence of insemination and pregnancy in the F 1 rats was unrelated to the dose of CPZ to which the Fo parents were exposed, with no statistical difference in incidence from the control group. Autopsy findings on the 14th day of pregnancy. As

shown in Table 5, the number of myometrial glands and the number of live fetuses were not different from the control group. The weight of the live fetuses from mating rats given 12.5, 25, and 100 mg/kg CPZ was significantly lower than that of the control group. No abnormalities were noted in gross observations of the uterus and the ovaries.

F2 generation Table 6 summarizes the date of birth of the F 2 rats

Table 4. Insemination and pregnancy data for F1 rats from parents (Fo) e x p o s e d to chlorpromazine for 9 weeks before breeding

Table 3. Data for pups (F1) from parental rats (Fo) exposed to chlorpromazine for 9 weeks before breeding Chlorpromazine, mg/kg/day (Mean ± SE) Observation N of dams (Fo) Length of pregnancy (days) Live pups/litter Average pup weight at hour 48 (g) Survival of pups at day 22 (%)

0 4 23.5 +--0.3 13.0 ±0.9 6.38 ±0.05 93.8

12.5

25

6 4 22.5 22.0 -+-0.4 ---0.7 12.0 11.3 -+0.6 -+0.9 6.42 6.04* -+0.03 -+0.04 95.8

100.0

50

100

5 22.4 ---0.7 11.4 ---0.9 5.65* ±0.36

4 22.0 ±0.6 9.0* ±0.9 5.48* ±0.36

80.0*

81.3"

*Significantly different from the 0 mg/kg group, P < 0.05.

Chlorpromazine, mg/kg/day Observation N of rats (F1) Male Female Insemination (%)

0

12.5

25

5 10 100.0

5 14 92.9

5 14 92.9

50 5 14 100

100 5 10 100

N of female rats inseminated/N of female rats paired with males (10/10) (13/14) (13/14) (14/14) (10/10) Pregnancy (%) 100 92.3 92.3 92.9 90.0 N of pregnant rats/ N of female rats inseminated

(10/10) (12/13) (12/13) (13/14) (9/10)

Breeding effect of chlorpromazine • K. S~ac-o

weaning period increased in rats descended from the Fo rats exposed to CPZ, as shown in Figure 3.

Table 5. Autopsy findings on day 14 of pregnancy in F~ rats from parents (Fo) exposed to chlorpromazine for 9 weeks before breeding

Wet weight of organs

Chlorpromazine, mg/kg/day (Mean -+ SE) Observation N of dams (F 0 MyomeWial glands/ litter Live fetuses/litter Average live fetal weight (g)

0

12.5

5 11.0 ___1.5 10.4 __.1.4 2.56 _0.01

6 10.8 __.0.5 10.2 __-1.1 2.48* ±0.03

25

50

The major organs in rats of each generation were weighed, and the ratios of wet weight of organs to body weight were calculated. The ratios, furthermore, were compared with those of the control group in order to evaluate the effect of CPZ on wet weight of organs. Administration of less than 50 mg/kg of CPZ produced no difference in the wet weight of major organs in male and female rats of each generation compared to the control group. With the administration of 100 mg/kg CPZ, as shown in Table 7, autopsy of the F o males at 21 weeks of age revealed an increased wet weight of the liver, kidney, and testis when compared to the control group, especially with a marked increase in the weight of the adrenal, pituitary, and prostate. Autopsy of the F o females at 24 weeks of age revealed an increased wet weight of the kidney, adrenal, and ovary, with a marked decrease in the weight of the pituitary and uterus. These tendencies were also noted in the weight of the major

100

6 7 10.8 9.7 ___0.8 --- 1.5 9.5 8.8 ±0.8 +--1.0 2.46* 2.45 ___0.02 ±0.05

4 10.5 ---0.8 9.3 ±1.0 2.23* ±0.06

*Significantly different from the 0 mg/kg group, p < 0.05.

(the length of pregnancy of the F~ females), the pup weight, and the survival on the 22nd day after birth. These values were not different from the corresponding values in the control group, and there was no correlation to the dose of CPZ to which the F o ancestors were exposed. Gross observations of the F 2 pups revealed no abnormalities. The body weight of the F 2 pups up to the

160

33

F2 pup

F~ pup

140

~

120

f~ "~ 1 0 0

EC°0r°aznei!!i!!

80

=

".~

.

50 mg/kg 100 mg/kg

60 2,

7,

= 14

2=1

Days

I

I

I

2

7

14

o

21 Days

Fig. 3. Rates of weight gain of the F 1 and F2 pups of ancestors (Fo) exposed to chlorpromazine for 9 weeks before breeding. *Significantly different from the values of control groups (0 mg/kg), p < 0.05.

34

Reproductive Toxicology

Table 6. Data for pups from F 1 rats whose parents (Fo) were exposed to chlorpromazine for 9 weeks before breeding Chlorpromazine, mg/kg/day(Mean _+ SE) Observation

0

12.5

25

50

100

N of dams (F1) 5 6 6 6 5 Length of pregnancy 22.2 22.6 22.4 22.6 22.4 (days) -+0.4 ---0.3 ---0.5 ---0.4 __-0.2 Live pups/litter 10.0 9.0 10.3 9.1 9.4 ---0.8 ---0.5 ---0.9 ---0.6 __-0.9 Average pup weight 6.56 7.41 6.62 6.63 6.40 at hour 48 (g) ---0.63 --+0.45 -+0.23 -+0.20 ---0.27 Survival of pups at day 22 (%) 97.5 92.9 96.4 96.4 95.0

organs of the F~ males and females at the weaning period. No difference from that of the control group was noted in the wet weight of the major organs of the F 1 males at 13 weeks of age, of the F1 females at 17 weeks of age, or of the F 2 males and females 3 weeks after birth. DISCUSSION CPZ has been used in the field of psychiatry for about 40 years. Studies on the reproductive effects of this drug have primarily relied on the tests for fetal toxicity (2, 3, 5, 6). Tests of influence on multiple subsequent generations have been reported only by Itoh et al. (6,7) and Fujii (8), In the present study, CPZ was administered daily to male and female rats for 9 weeks prior to mating. The effects of CPZ on reproduction in 3 generations were evaluated. The estrous cycle in mature female rats is usually 4 to 5 days (10) but the estrous cycle in the female Sprague-Dawley strain rats used in the present study was 4 to 10 days. The estrous cycle of the Fo rats was prolonged depending on the dose of CPZ to which they were exposed. These results were in agreement with the studies of Brraclough and Sawyer (11), suggesting a prolongation of estrous cycle in rats due to CPZ. Consequently, the prolongation of the estrous cycle by CPZ appears to be correlated with changes in the incidence of insemination and pregnancy in Fo rats that had been administered CPZ. Estrous cycles of the F 1 females were 4 to 10 days, with no correlation to the CPZ which had been dosed in the Fo generation. The prolongation of the estrous cycle by CPZ is induced only in the parent generation exposed to CPZ and appears to have no influence on the next generation. The incidence of insemination and pregnancy in the F o rats showed a tendency to decrease depending on the dose of CPZ. This was especially seen in the mating pairs of the F o rats treated with a high CPZ dose(100

Volume 4, Number 1, 1990 mg/kg). In the mating groups of rats given a low dose of CPZ (12.5 and 25 mg/kg), the incidence of insemination was of a magnitude similar to that in the control group. The present results, that CPZ exerts influence on the incidence of pregnancy, are supported by the report of Hoekstra et al. (12) on the inhibition of ovulation by CPZ, with a decrease in the degree of inhibition by repeated administration of CPZ (12,13), and also by the report of Sliwa (14,15) on the decrease in the number and motility of mouse spermatozoa by CPZ, with a reduced fertilization capability. In a study to evaluate the influence of the benzodiazepine series of anti-anxiety drugs on reproduction, administration of the drug to both male and female rats inhibited the mating performance of mice (16) and pregnancy in rats (17), but drug administration in male rats alone failed to influence reproduction (18). CPZ decreased the incidence of insemination and pregnancy in a fashion similar to that produced with the benzodiazepine series drugs, but the incidence was modified by the dose and the duration of administration. Since the decrease in the incidence of insemination and pregnancy due to CPZ was not found in the F 1 generation, CPZ was found not to influence insemination and pregnancy in successive generations. The length of pregnancy of the Fo and F~ females was 22 to 23 days, as was the length in the control group. These figures are in agreement with previously reported pregnancy length in rats (10). Natural delivery of litters was normal regardless of the duration of CPZ administration. Autopsy of the Fo and F 1 rats revealed the same number of myometrial glands per litter with no correlation to the dose or the duration of CPZ administration. The number of live fetuses per litter depended on the dose of CPZ except for the F 1 generation. The weight of the F~ and F 2 fetuses also was decreased depending on the dose of CPZ. CPZ inhibited body weight gain of the F o rats in a dose-dependent fashion. The body weight of the F~ pups 48 h after birth was also inhibited depending on the dose of CPZ to which the F o rats were exposed. The growth of the F~ pups following the 14th day after birth was favorable; however, the low-birth-weight newborn rats had lower body weight than the control rats for about 3 weeks after the weaning. The body weight of the F 2 pups 48 h after birth was not different from that in the control group, and the growth of the F 2 pups was favorable. The body weight of the F 2 pups showed a significantly greater increase than the control group. At 48 h after birth, the number of live pups per litter decreased depending on the dose of CPZ to which the Fo rats were exposed. The death of the newborns occurred mostly within the first several days after birth, and none died after the 14th day of life. Survival of the F 1 r a t s at weaning decreased in each group and depended on the dose of CPZ to which the Fo rats were exposed. Survival of the F 1 rats from the mating group of rats who received

35

Breeding effect of chlorpromazine• K. S~ac,o Table 7. Comparison of organ weights in chlorpromazine (100 mg/ kg)-exposed rats (Fo) and their offspring (FI) with those in control rats

Generation Vo 10 Males 4 Females

Weeks Testis* Prostate* of or or age Liver Kidney Adrenal Pituitary ovary** uterus** 21 24

+ _

3 3

-+ -

+ ++

++ ++

++ -

+ ++

++ -

+ -

+ -

+ +

+ -

FI 10 Males 10 Females

___ -

Marks of * and ** = male and female, respectively. The effect of chlorpromazine on organ weights was evaluated by the following formula: A (control) = organ weight/bodyweight B (treated) = organ weight/bodyweight C % = 100 × (B-A)/A Marks of - , --, + and + + indicate the values when C value is less than - 6%, between -5% and +5%, between +6% and 10%, and more than +11%, respectively.

100 mg/kg CPZ was 81.3%. Survival of the F2 pups at weaning failed to correlate with the dose of CPZ. It is well known that when the number of newborns per litter is small the growth of the young is more rapid, with a marked increase in body weight (10). When the number of newborns and their body weight 48 h after birth were similar to controls, the subsequent growth of the young was increased and body weight gain was significantly greater than that in the control group. This appears to be due to the influence of CPZ. CPZ, however, is known to increase prolactin secretion from the anterior pituitary, causing mammary hypertrophy and facilitating lactation (19,20). The relationship between a possible increase in CPZ-induced milk secretion and the body weight increase in the newborn was unclear. McClain and Hoar (17) have suggested that the increase of newborn death due to flunitrazepam is due to central nervous system depression of the mother animal by drug administration during pregnancy, leading to indifference of the mother animal to the newborn. In the present study, CPZ was administered prior to pregnancy, so that a similar conclusion of a relationship between newborn death and CPZ cannot be drawn. The answer remains a future research problem. However, the apparent influence of CPZ administered to the F 0 generation prior to insemination on the mortality of both F 1 and F 2 generations is of interest. The influence of CPZ on wet weight of organs was not significant following the administration of less than 50 mg/kg of CPZ in the F o, F~, or F z generations. After administration of 100 mg/kg CPZ, however, the wet weight of the adrenal, pituitary, and prostate glands, especially in the F o males, increased. The weight of the kidney, adrenal gland, and ovary in the F o females

increased, but that of the pituitary gland and the uterus decreased. Hafs and Williams (21) administered CPZ to goats for 8 weeks with no remarkable influence on the reproductive organs; these findings differ from the results we have obtained in rats. In tests for fetal toxicity in rats, CPZ is reported to cause cleft palate (2,3,10,22,23); however, cleft palate can occur in the fetuses when food and water are withheld from the mother (23). Although CPZ decreases maternal food and water intakes, whether or not CPZ has a primary teratogenic effect has not been decided. In the present study, autopsy of the rats in the F o, F 1, and F 2 generations revealed no abnormalities in gross observations. There was a decrease in spontaneous motor activity, body weight, and food and water intake noted in both male and female F o rats. This decrease was not found in the F 1 and F 2 generations except for the influence of CPZ on body weight in the F t generation. There are some reports regarding the relationship between the decrease of body weight and the inhibition of spermatogenesis. Krause et al. (24), Sobhon et al. (25), and Bint et al. (26) reported that both the body weight and spermatogenesis were inhibited in the experiments involving retinol deficiency in rats, vitamin A deficiency in rats, and anti-GnRH in monkeys, respectively. On the other hand, Wilson et al. (27) reported that in an experiment in which broiler birds were fed a low-protein diet, body weight was decreased by the diet, but spermatogenesis was unaffected. In the present study, although CPZ produced a decrease in body weight and incidence of insemination and pregnancy, it is difficult to prove whether the decreased body weight seriously inhibits spermatogenesis. However, from the above described results and the report of Sliwa (14,15), who showed that CPZ inhibited spermatogenesis, it is

36

Reproductive Toxicology

suggested that CPZ may produce an inhibition of both body weight and spermatogenesis. Blepharitis also occurred in the F 0 males and females, although it was more prominent in males than in females, in agreement with the report by Itoh et al. (28). These results suggest that CPZ exerts an inhibitory influence on the growth of the fetuses and the newborns from the F o and F 1 generations when it was administered to both male and female F 0 rats before breeding, resulting in reproduction inhibition.

Volume 4, Number 1, 1990

12.

13.

14. 15. 16.

17.

- - This work was supported by a research grant from the Shionogi Seiyaku Company. The author wishes to thank Professor T. Itoh in the Department of Pharmacology, School of Dentistry, Iwate Medical University, for his excellent technical direction and kind advice.

18.

REFERENCES

19.

1. Gilman AG, Goodman LS, Gilman A. The pharmacological basis of therapeutics. 6th ed. New York: Macmillan; 1980:391-418. 2. Beall JR. A teratogenic study of chlorpromazine, orphenadrine, perphenazine, and LSD-25 in rats. Toxicol Appl Pharrnacol. 1972;21:230~. 3. Daily JW. Effects of maternal chlorpromazine on offspring nervous system development. Neuropharmacol. 1978;17:583-7. 4. Itoh T. Our clinical reflection on toxic reactions and side-effects induced by phenothiazine drugs. J lwate Med Assoc. 1976;28: 259-70. 5. Robertson RT, Majka JA, Peter CP, et al. Effects of prenatal exposure to chlorpromazine on postnatal development and behavior of rats. Toxicol Appl Pharmacol. 1980;53:541-9. 6. Itoh T, Ando F, Seki M, et al. Effect of chlorpromazine on the reproduction in rats. Jap J Pharmacol. 1976;26(Suppl):90. 7. Itoh T, Murai S, Nakaya S. Effects of chlorpromazine administered to first generation rats on reproduction of the next generation. Jap J Pharmacol. 1978;28(Suppl):164. 8. FujiiT. Strategies for the assessmentoftheeffect of matemal drug exposure on the offspring. J Toxicol Sci. 1987;12:558-67. 9. Park AW. Aspects of oral development in relation to litter size in rats. Acta Anat. 1968;70:403-15. 10. Takagi H, Takano K, Kikuchi Y, et al., eds. Special toxicity of drugs. I: Fetal and genetic toxicity. Tokyo: Nankodo; 1975:1-35, 195-7. 11. Brraclough CA, Sawyer CH. Induction of pseudopregnancy in the

20.

Acknowledgements

21. 22. 23. 24.

25.

26.

27. 28.

rat by reserpine and chlorpromazine. Endocrinol. 1959;65:563571. Hoekstra A, Yih TD, Joosten HFP, et al. The effects of chlorpromazine, mianserine and ORG GC94 on reproductive function in the rat. Drug Chem Toxicol. 1984;7:11-22. Kogo H, Iida H, Inazu N, et al. Inhibition of the formation of 13,14-dihydroprostaglandin F2-alpha induced by chlorpromazine in rat ovary. Prostaglandins Leukotriens Med. 1986;22:11-20. Sliwa L. Gametogenesis in chlorpromazine-treated BALBc mice. Folia Biol. 1984;31:303-14. Sliwa L. Motility and fertilization capability of mouse and toad, chlorpromazine-treated spermatozoa. Folia Biol. 1984;31:381-92. Guerriero FJ, Fox KA. Benzodiazepines and reproduction of Swiss-Webster mice. Res Comm Chem Path Pharmacol. 1976; 13: 601-10. McClain RM, HoarRM. The effect offlunitrazepam on reproduction in the rat. The use of cross-fostering in the evaluation of postnatal parameters in rat reproduction studies. Toxicol Appl Pharmacol. 1980;53:92-100. Wilkinson M, Moger WH, Grovestine D. Chronic treatment with valium (diazepam) fails to affect the reproductive system of the male rat. Life Sci. 1980;27:2285-91. Itoh T. Dose-related response of mouse mamma to some phenothiazine drugs. J Toxicol Sci. 1978;3:147-52. Meltzer HY, Fang VS. The effect of neuroleptics on serum prolactin in schizophrenic patients. Arch Gen Psychiat. 1976;33: 279-86. Hafs HD, Williams JA. The effects of prolonged chlorpromazine administration on the reproductive organs in the male goat. Am J Vet Res. 1964;25:523-8. Tanimura T. Problems on testing drugs for their effects on offspring. Lyakuhin Kenkyu. 1974;5:285-94. Szabo KT, Brent RL. Species differences in experimental teratogenesis by tranquillizing agent. Lancet. 1974;1:565. Kranse RF, Beamer KC, McCormick AM, et al. The effect of retinol and retinoic acid on physiological and biochemical changes in retinol-deficient rats. Br J Nutr. 1975; 33:73-85. Sobhon P, Mitranond U, Tosukhowong P, et al. Cytological changes in the testes of vitamin A-deficient rats. II. Ultrastructural study of the seminiferous tubules. Acta Anat. 1979;103:169-83. Bint AF, Weinbauer, GF, Nieschlag E. Acute and chronic effects of a gonadotropin-releasing hormone antagonist on pituitary and testicular function in monkeys. J Endocrinol. 1985;104:345-54. Wilson JL, McDaniel GR, Sutton CD, et al. Semen and carcass evaluation of broiler breeder males fed low protein diets. Poult Sci. 1987;66:1535-40. Itoh T, Murai S, Yoshida H, et al. Alteration in the effects of repeated administrations in rats of various combination doses of propericiazine and haloperidol. Jap J Neuropsychopharmacol. 1981 ;3:485-99.