Hormonal induction of ovulation stimulates atresia of antral follicles in a vespertilionid bat, Scotophilus heathi

Scotophilus heathi is a seasonally monoestrous subtropical vespertilionid bat found at Varanasi, India. Although the antral follicles remain present in the ovaries of S. heathi from November till March, ovulation is delayed in this species until early March. In order to understand the mechanism of ovulation suppression during this period of delayed ovulation, the effects of human chorionic gonadotropin (hCG), pregnant mare's serum gonadotropin (PMSG), follicle stimulating hormone (FSH) and gonadotropin releasing hormone agonist (GnRH agonist) on ovarian morphology and steroid concentration were investigated. Hormonal treatments were given as a single i.p. dose 24 h after capture. The bats were sacrificed 48 h after the injection. Treatment with hCG, PMSG, FSH and GnRH agonist failed to induce ovulation in S. heathi, although these hormones produced a high degree of ovarian stimulation. The administration of hCG and PMSG induced ovarian enlargement, intense hyperemia, marked changes in the interstitial cells (ICs), development of several antral follicles and a varying degree of abnormalities in the oocytes of most of the antral follicles. In the bats treated with hCG, PMSG and GnRH agonist, androstenedione concentration increased significantly to extraordinarily high levels, whereas estradiol concentration decreased. Administration of FSH caused regression of ICs and pyknosis of granulosa cells in the majority of antral follicles. FSH did not enhance androstenedione concentration. The results of the present study suggest that the failure of hormonal treatments to induce ovulation during the period of delayed ovulation might be due to a seasonal desensitization of ovarian follicles in S. heathi. The hormonal treatment instead stimulated the ICs to produce a high level of androstenedione resulting in atretic changes of the antral follicles.     

Seasonal adiposity and androstenedione production as a possible mechanism for asynchronous reproductive activity between males and females of vespertilionid bat, Scotophilus heathi

Relationship between changes in the body weight and circulating androstenedione concentration was examined in order to find out their involvement in asynchronous gametic cycle of male and female S. heathi. Body weight of both male and female bats increased from September due to accumulation of white adipose tissue and reached to a peak in November. The body weight declined gradually due to consumption of adipose tissue during December and January reaching basal level in February. A significant correlation between body weight and serum androstenedione concentration was observed. Correlated with maximum body weight, peak androstenedione level was found during November in both male and female bats. Peak androstenedione level during November may be responsible for stimulating spermatogenesis in the male, but may suppress follicular growth and ovulation in the female, thus resulting in asynchronous gametic cycle of male and female S. heathi.     

Changes in serum leptin, insulin, androstenedione and luteinizing hormone during ovarian cycle in the bat, Taphozous longimanus

Detailed reproductive pattern and associated endocrine characteristics have been documented in only a few species of order Chiroptera. The aim of the present study was to examine the changes in body weight, serum insulin, leptin, androstenedione and luteinizing hormone (LH) concentrations during annual ovarian cycle in the sheath-tailed bat, Taphozous longimanus. Bats were sampled over three years. Leptin, a satiety hormone produced primarily by adipose tissue, provides information to feeding center of the brain about nutritional status, fat mass, appetite and energy expenditure. The circulating concentration of leptin begins to increase from October and attains a peak in December. The peak serum leptin concentration coincides with body weight in November before winter dormancy in December. The serum leptin levels dissociate from body weight during December. The other peaks of serum leptin levels coincide with late stages of the two successive pregnancies. The serum insulin concentration begins to increase from September and attains a peak during December. The insulin concentration remains low from January to August. The circulating androstenedione concentration begins to increase in October, reaching a peak in December. This increase in androstenedione concentration correlated with the period of heavy accumulation of abdominal fat and increase in body weight. There was a sharp decline in androstenedione concentration and body weight in January. The serum LH shows peaks, in November, coinciding with the peaked body weight, the other peaks in January and May, coinciding with ovulation for the two successive pregnancies. The high leptin and insulin levels might be responsible for the maintenance of reproductive response and gonadal function during adverse environmental condition in the winter, while high androstenedione, and associated body weight along with LH might be responsible for maintaining basal gonadal function. We conclude that high leptin, androstenedione and insulin serve, as signal for the reproductive functions in that sufficient body fat are available to meet the caloric demands and maintain normal function during adverse winter conditions.     

Relationship between corticosterone and body weight,androstenedione and insulin during the period of delayed ovulation in a vespertilionid bat,Scotophilus heathi

The aim of the present study was to evaluate the relationship between corticosterone, body weight, insulin and androstenedione in order to understand the role of adrenal in contributing hyperandrogenism during delayed ovulation in S. heathi. The circulating corticosterone concentration in female S. heathi showed significant seasonal variation. The peak corticosterone concentration observed during August-September coincides with increased feeding activities in S. heathi. The present study noted a seasonal variation in relationship of corticosterone with insulin and androstenedione in S. heathi. An inverse relationship of corticosterone with insulin and androstenedione was found during August to December, but not during January to May. A seasonal variation in the effect of adrenocorticotropic hormone (ACTH) on adrenal corticosterone production in vitro was observed during reproductive cycle. Corticosterone production in vitro by adrenal declined significantly as compared to the control during quiescence in September. The finding suggests that adrenal attained the peak responsiveness to ACTH during September. ACTH significantly enhanced the androstenedione production by the adrenal in vitro during December, when the circulating androstenedione was also high in S. heathi. This suggests that the adrenal may also contribute to hyperandrogenism during the period of delayed ovulation in S. heathi. Further studies are required to reveal the unique pattern of seasonal relationship between corticosterone, insulin and androstenedione in S. heathi.     

Year round plasma leptin and androgen concentrations in a tropical bat

The detailed reproductive patterns and their associated endocrine characteristics have been documented only for a few species of bats. The objective of this study was to examine seasonal changes in plasma concentrations of leptin and compare it with the changes in body mass, circulating concentrations of testosterone, androstenedione and its correlation with prolonged survival of sperm during winter dormancy in the male sheath-tailed batTaphozous longimanus Hardwicke, 1825. Six bats were captured every month for three consecutive years during 2002 to 2005 from Varanasi, a subtropical part of India. The changes in the body mass were positively correlated with circulating concentration of leptin. Leptin concentration reached a peak (14 ng/ml) in November coinciding with peak body mass. Leptin levels declined during other months of the year except for a rise in March and August. Plasma leptin was positively correlated with androstenedione concentration, but did not show significant correlation with testosterone level. We noticed a significant increase in testosterone secretionin vitro in response to leutinizing hormone (LH) stimulation. However, we did not notice any increase in testosterone or androstenedione secretionin vitro in response to leptin stimulation. Plasma leptin concentration did not show any correlation with testis mass in this study. The higher concentration of testosterone and androstenedione may be responsible for the prolonged survival of sperm in the epididymidies and higher levels of leptin in November may be responsible for maintaining reproductive function during winter dormancy. We suggest that inT. longimanus, higher leptin concentrations in November may be responsible for the gonadal recrudescence and reproductive response during winter dormancy is modified by energy availability and by changing leptin concentrations during this period.     

Mechanism of delayed ovulation in a vespertilionid bat, scotophilus heathi: role of gonadotropin, insulin, and insulin-like growth factor-1

The aim of this study was to evaluate the effects of luteinizing hormone (LH), follicle-stimulating hormone (FSH), insulin, and insulin-like growth factor-1 (IGF-1) on ovarian androstenedione synthesis to understand the mechanism responsible for delayed ovulation in Scotophilus heathi. We found that LH stimulated a dose-dependent increase in androstenedione synthesis by the ovary in vitro. This study also showed a clear seasonal variation in the ability of the ovary to produce androstenedione in vitro in response to LH and FSH stimulation. In response to LH and FSH, maximum quantities of androstenedione were produced during recrudescence in November. The same doses of gonadotropins during the preovulatory period in February stimulated comparatively low androstenedione secretion by the ovary. On the basis of these data, we suggest that in S. heathi, ovarian responsiveness to LH and FSH peaks during recrudescence. This study also showed a seasonal variation in the effects of insulin and IGF-1 on ovarian androstenedione production in vitro. Peak ovarian responsiveness to insulin and IGF-1 was observed during quiescence in September. It is hypothesized that increased insulin/IGF-1 sensitivity during September may be responsible for increased responsiveness to LH. Increased LH release, if coincident with the period of enhanced ovarian responsiveness to LH, may result in the excessive androstenedione production responsible for delayed ovulation in S. heathi.     

Reproduction in Indonesian sheep and goats at two levels of nutrition

Oestrus and ovulation were recorded for eight oestrous cycles after sheep and goats were put onto maintenance or supramaintenance levels of nutrition. Oestrous cycles were longer and more variable in goats than in sheep (20.3 ± 0.3 and 17.4 ± 0.2 days respectively), but in neither species was the incidence of oestrus affected by level of nutrition. There were no significant differences between sheep and goats in average body weight or mean ovulation rate. Animals fed at a maintenance level showed no significant changes in average body weight (19.0 ± 0.3 kg) or mean ovulation rate (1.69 ± 0.01). At the supramaintenance level, on the other hand, mean body weight increased progressively to a maximum of 24.8 ± 0.5 kg, and mean ovulation rate increased to a maximum of 2.65 at the sixth oestrus after the beginning of supramaintenance feeding. The effect of supramaintenance feeding on ovulation rate was reflected in subsequent lamb and kid production.     

Seasonal changes in circulating steroid concentration and their correlation with the ovarian activity in the female Indian sheath-tailed bat, Taphozous longimanus

The relationship between ovarian activity and circulating steroid concentration was studied in the female sheath-tailed bat, Taphozous longimanus. T. longimanus breeds twice in rapid succession during the year at Varanasi, India. Ovarian recrudescence was observed during September, and antral follicles were first observed during the month of October. Circulating androstenedione concentration showed an increase beginning in October, reaching a peak in December. This increase in androstenedione concentration correlated with the period of heavy accumulation of adipose tissue and increase in body mass. Antral follicles grow slowly during the period of high circulating androstenedione concentration from October to December. There was a sharp decline in androstenedione concentration during January. Simultaneously with the decline in androstenedione concentration, a sharp increase in size of the antral follicle and circulating estradiol concentration was noticed. Soon thereafter, one of the follicles ruptures, followed by fertilization and the commencement of the first pregnancy. During the second pregnancy, antral follicles first appeared during late pregnancy in March in the contralateral ovary that lacked the corpus luteum. One follicle developed rapidly and quickly attained a preovulatory stage in April. This is reflected in a sharp increase in estradiol concentration during this period. Ovulation was observed immediately following the first pregnancy in May. During this period, androstenedione concentration remained low. The results of the present study suggest that high androstenedione concentration during October to December (winter dormancy) may be responsible for slow follicular development and delays ovulation in T. longimanus. It is further hypothesized that the geographical variation in reproductive pattern of T. longimanus could be due to variation in the duration of fat storage and associated changes in the androstenedione concentration.     

Pattern of ovarian protein synthesis and secretion during the reproductive cycle of Scotophilus heathi: synthesis of an albumin-like protein

Ovarian proteins synthesized de novo and secreted in vitro were investigated during different stages of the reproductive cycle of Scotophilus heathi. We found increased ovarian protein synthesis and secretion during the recrudescence and the preovulatory periods, coinciding with two peaks of follicular development and steroidogenesis. The ovaries synthesized protein at a low rate during quiescence and the late phase of delayed ovulation. In vitro analysis using ³⁵S-methionine incorporation showed increased synthesis of 66 kDa protein during delayed ovulation, but secretion of this protein declined markedly during the preovulatory period. N-terminal sequencing of the 66 kDa protein showed that it shares 70% homology with human serum albumin. Immunocytochemistry indicated that this protein is found primarily in the granulosa cells of the follicles. Whether the increase in albumin production by the ovaries during delayed ovulation is associated with a hyperinsulinemic and hyperandrogenic condition requires further investigation.     

Seasonal adiposity, correlative changes in metabolic factors and unique reproductive activity in a vespertilionid bat, Scotophilus heathi

The aim of this study was to determine the effect of changes in body mass, fat reserves and feeding activity on circulating levels of lipids, glucose, protein and metabolic hormones in a vespertilionid bat, Scotophilus heathi. Furthermore, the relationship between changes in metabolic factors and hormones with the unique reproductive features of female S. heathi was also examined. The results of this study showed annual variation in body mass, fat reserve and feeding activity, which correlated significantly with circulating levels of lipids, protein and metabolic hormones. Increased corticosterone level during September-October in S. heathi promotes increased feeding activity, which in turn induces hyperinsulinemia in S. heathi during November. Hyperinsulinemia together with low body temperature in November facilitates fat accumulation in bat. Coinciding with the period of fat accumulation raises serum leptin level, which has been demonstrated to suppress ovarian activities thus causing delayed ovulation in S. heathi. Circulating levels of lipids were high during winter dormancy, which may provide energy to stored sperms. The study thus suggests that the unique reproductive features of female vespertilionid bat are strongly linked to fat deposition. J. Exp. Zool. 309A:94-110, 2008. (c) 2008 Wiley-Liss, Inc.     

Melatonin modulates glucose homeostasis during winter dormancy in a vespertilionid bat,Scotophilus heathi

The role for melatonin in glucose homeostasis and insulin resistance is not very clear and has recently been an active area of investigation. The present study investigated the role of melatonin in seasonal accumulation of adipose tissue in Scotophilus heathi, with particular reference to its role in glucose homeostasis and development of insulin resistance. The circulating melatonin levels correlated positively (p < 0.05) with the changes in body mass due to fat accumulation and circulating insulin level, but correlated negatively with the blood glucose level in S. heathi. The bats showed high circulating blood glucose levels and impaired glucose tolerance during the period of fat deposition suggesting insulin resistance condition which improves after winter when most of the fat has been utilized as a metabolic fuel. The high circulating melatonin levels during the period of maximum body fat at the beginning of winter prepare the bats for winter dormancy by modulating the glucose homeostasis through affecting blood glucose levels, muscle and liver glycogen stores, insulin receptor and glucose transporter 4 (GLUT 4) expression. This is also confirmed by in vivo study in which melatonin injection improves the glucose tolerance, increases muscle insulin receptor and GLUT 4 expression, and enhances glucose clearance from the blood. The results of present study further showed that the effect of melatonin injection on the blood glucose levels is determined by the metabolic state of the bats and may protect from decrease in blood glucose level during extreme starvation, however, melatonin when injected during fed state increases glucose clearance from the blood. In summary, the present study suggested that melatonin interferes with the glucose homeostasis through modulating intracellular glucose transport and may protect bats from hypoglycemia during winter dormancy.     

Seasonal changes in the content of lipids and photosynthetic pigments in a brown alga Saccharina cichorioides

Seasonal changes in the contents of lipids and photosynthetic pigments (PSP) were investigated in a brown alga Saccharina cichorioides Miyabe (Phaeophyceae, the family Laminariaceae). The content of lipids varied from 0.27 to 0.60% of the algal fresh weight. The content of glyceroglycolipids (GL) was much greater in the time of spore formation (June–July and September–October), phospholipids — in the spring and in September–October, and the content of neutral lipids — in the spring and in November. In the period of spore release (August and October), the level of GL and polyunsaturated fatty acids (PUFA) sharply decreased. A high level of PUFA was observed from March to July and in November. In August and October, the same as in the spring, the proportion of saturated fatty acids (FA) was great. The content of chlorophylls from March to November varied from 20.3 to 26.9%, and the level of carotenoids — from 10.7 to 16.1%. Total content of PSP was relatively high in March and in August–September. Free sterols accounted for 3.4–7.3% of total lipids; their proportion was greater in spring than in summer and autumn.     

Ovarian steroidogenesis during follicular maturation in the domestic fowl (Gallus domesticus)

The steroidogenic potential of various physiological compartments within the ovary of the hen were examined using in vitro systems. Three-hour incubations of individual whole small follicles (less than 1 mm-1 cm) or 100,000 collagenase-dispersed theca cells of the five largest ovarian follicles (F1-F5) were conducted in 1 ml of Medium 199 at 37 degrees C in the presence and absence of luteinizing hormone (LH) (0.39, 0.78, 1.56, 3.13 and 6.25 ng), progesterone (5 ng), and dehydroepiandrosterone (DHEA, 5 ng). Steroid output was measured by radioimmunoassay of incubation media. Progesterone was not produced by small follicles although they are a major source of DHEA and estradiol and a significant source of androstenedione. Output of DHEA, androstenedione and estradiol was highly stimulated by LH. The substrate for androstenedione and estradiol in small follicles is probably DHEA. Output of DHEA and androstenedione in theca cells of F2-F5 was stimulated by LH in a dose-related manner. A dose-response relationship between estradiol output and the concentration of LH in media was not apparent in theca cells from F2-F5. Steroidogenesis in theca tissue of large follicles occurs predominantly via the delta 4 pathway. The ability of these theca cells to metabolize progesterone to androstenedione is lost between 36 and 12 h before ovulation. Their ability to metabolize DHEA to androstenedione is still present 12 h before ovulation. Aromatase activity is significantly reduced between 36 and 12 h before ovulation. These data indicate that both large and small follicles can be stimulated by LH. The small follicles are the major source of estrogen. As the large yolky follicles mature, steroidogenesis shifts from the delta 5 to the delta 4 pathway. By 12 h before ovulation, the F1 follicle has lost the ability to convert progesterone to androstenedione. The inability of the largest ovarian follicle to convert progesterone to androstenedione contributes at least in part to the preovulatory increase in the plasma concentration of progesterone that generates the preovulatory LH surge by positive feedback.     

Observations on the female reproductive cycles of captive Asian yellow pond turtles (Mauremys mutica) with radiography and ultrasonography

The annual reproductive cycle of 27 female Mauremys mutica was observed by radiography and ultrasonography from April 2006 to August 2007. Radiography was used to monitor clutch size and ultrasonography was used to monitor changes in the ovarian follicles. The follicles started to enlarge in September and became preovulatory in January. The mean maximum follicle diameter of ovulation was 18.30±1.44 mm, and ovulation occurred from March through August. Eggs were laid between April and August. Turtles entered latent period in early August and the maximum follicular size was at a low of 13.22±2.36 mm in late September. The vitellogenesis of the next reproductive cycle began in October. The 24 adult females laid 56 clutches containing a total of 227 eggs. Average clutch size was 4.05 eggs (range 1–8) and there were 2.33 clutches (range 1–4) per female. Egg shell images were first observed on the sixth or seventh day after ovulation. The oviductal period averaged 6.9 weeks (range 2–16 weeks) on the first clutch, 3.4 weeks (range 2–8 weeks) on the second, and 2.75 weeks (range 2–6 weeks) for the third. Radiography and ultrasonography are non‐invasive and convenient methods to evaluate the reproductive cycle of female M. mutica. They should be applicable to other turtles and should greatly enhance knowledge of reproductive physiology. Zoo Biol 29:50–58, 2010. © 2009 Wiley‐Liss, Inc.     

The lifespan of the cladoceran Ceriodaphnia affinis Lilljeborg in a laboratory culture

The lifespan of the cladoceran Ceriodaphnia affinis Lilljeborg in a laboratory culture has been determined. The lifespan decreases in June and from November to December and it increases from February to March and from August to September. Minor changes in temperature over the year (21±3°C) have no direct effect on the lifespan of the animals. The average lifespan proves to be a more variable parameter than the maximum lifespan.     

The influence of azaperone treatment at weaning on reproductive function in sows: ovarian activity and endocrine profiles during the weaning-to-ovulation interval

Azaperone can reduce stress caused by weaning and relocation of breeding sows, but its effects on reproductive processes are still poorly understood. The primary aim of this study was to describe and compare the endocrine and ovarian activity in ultrasonographically monitored second parity sows, with or without azaperone treatment at weaning (2 mg/kg BW i.m.). The intervals from weaning to the onset of estrus and ovulation were both greater (P<0.05) in azaperone-treated (n=12) than in control sows (n=12) by ~12 h. Mean daily growth rates of identified antral follicles were less (P<0.05) in azaperone-treated than in control sows (1.08±0.17 v.1.23±0.18 mm/day; mean±SD) and treated animals exceeded (P<0.05) controls in the mean ovulation rate (13.7±1.3 v. 12.6±1.2). A transient suppression of cortisol release was observed in the treatment group (at 10 and 30 min after azaperone injections) but circulating cortisol concentrations were variable in both groups of sows for the remainder of the study. The preovulatory rise in LH and estradiol secretion was delayed (P<0.05), and the duration of the LH surge was greater (P<0.001) in azaperone-treated sows compared with their control counterparts. The amplitude of episodic fluctuations in serum cortisol concentrations was correlated with the number of stillborn piglets in control sows (r=0.63, P=0.04). The amplitude and concentration of the preovulatory rise in estradiol secretion were negatively correlated with ovulatory response and litter size (r=−0.63 to −0.82, P<0.05), whereas the time at which the LH surge ended was directly related to the number of live-born piglets (r=0.82, P=0.002) in azaperone-treated animals. The present results indicate that administration of azaperone at weaning had a profound effect on preovulatory LH secretion as well as growth kinetics and estrogenicity of ovarian antral follicles. However, the causative associations among various characteristics of the preovulatory LH discharge, ovarian and adrenal steroid secretion post-weaning, and reproductive variables in sows remain equivocal.     

The seasonal pattern of testosterone secretion in finn cross rams in Israel

Three adult Finn cross rams were bled at 30-min intervals for 7 h in March, May–June, August and November, and again at hourly intervals for 2 h after an intravenous injection of 50 μg of a GnRH analogue. Plasma testosterone concentrations were measured by radioimmunoassay. The mean testosterone level from 15 blood samples for each ram was highest in November and lowest in March. The episodic pulse pattern of testosterone secretion during the 7-h blood sampling periods was most evident, and more similar among rams in August and November, and less so in March and May.The mean testosterone concentration from the blood samples collected 1 and 2 h after GnRH injection was significantly higher in August–November than in March–June, but the ratio of the testosterone level after to that before injection was highest in March and lowest in November.     

Seasonal incidence of aphid,Amphorophora ampullata Bukton (Homoptera: Aphididae) on fern,Hypolepis polypodioides (Blume) Hook (Hypolepidaceae) from Western Himalaya

Seasonal incidence of aphid, Amphorophora ampullata on Hypolepis polypodioides was recorded throughout the year from November 2012 to November 2013 at weekly interval. Peak incidence of aphid was recorded during third week of November 2012 (25.94 ± 2.39 aphids/pinna), later the aphid population gradually decreased from December 2012 onwards and reached below threshold level during the last week of January (0.1 ± 0 aphids/pinna). The aphid population starts building up again from first week of February 2013 (1.6 ± 0.13 aphids/pinna) and attained its peak in the last week of August (32.17 ± 1.22 aphids/pinna) then decreased gradually in the first week of September (20.82 ± 4.70 aphids/pinna). Aphid densities again increased gradually from second week of September (21.62 ± 3.02 aphids/pinna) to November 2013 and reached maximum aphid densities during November (56.55 ± 4.34 aphids/pinna). Among weather parameters, aphid population showed significant positive correlation with relative humidity during morning hours.     

Summer diapause and winter quiescence ofCoccinella septempunctata (Col. Coccinellidae) in central Greece

Coccinella septempunctata (Coleoptera: Coccinellidae) in Greece is a multivoltine species. In the lowlands, all instars are abundant in spring becoming scarce from July until the end of the warm period of the year; they are absent in winter. In June, mostC. septempunctata adults migrate to mountainous aestivo-hibernation sites. Measurements of the duration of pre-oviposition period in females taken monthly from the summit of Mount Kitheron in central Greece (1993–1994) and transferred to laboratory conditions of high temperature (25°C), long day (16 hrs light/24 hrs), and presence of aphids, indicated that theC. septempunctata females were in diapause during July and August. The diapause gradually terminated from late August to late October and was followed by a period of quiescence extending from November to March of the following year. During the summer diapause, arrest of ovarian development was indicated by immaturity of the ovaries in all sampled females and the complete absence of vitellogenic resorption signs. Also, adults were found with enlarged fat bodies, and the median duration of preoviposition period in females transferred to the above laboratory conditions was 90 days in early July and 82 days in early August. During the period of winter quiescence, arrest of ovarian development was characterized in most samples by immaturity of the ovaries in all females and, in some samples, by the presence of a few females with signs of vitellogenic resorption. In winter, adults were found containing fat body reserves of different levels, and the median preoviposition period of females transferred to optimal breeding conditions was 29 days in early November and 16 days in mid January.     

Delayed sexual maturation in the female rat during chronic exposure to delta-9-tetrahydrocannabinol

delta-9-Tetrahydrocannabinol (THC), the major psychoactive component of marihuana, retarded sexual maturation in the pubertal female rat as indicated by a delayed appearance of estrus and ovulation. Twice daily intraperitoneal injections of 10 mg THC/kg body weight from 27 days of age until the first day of full vaginal cornification delayed the onset of estrus and the occurrence of the first ovulation by an average of 4.3 +/- 0.9 (S.E.M.) days in comparison with pair-fed, vehicle-treated animals (p less than 0.001). Ovulation during the first estrus occurred in 92% of the animals in each treatment group without a difference in the average number of ova ovulated. The delayed sexual maturation in animals treated with THC could not be attributed to reduced body weight gain since body weights in THC- and vehicle-treated groups were equivalent when vehicle-treated animals came into estrus. Delayed ovulation in THC-treated animals did not merely reflect the repeated blockade of ovulation from otherwise competent follicles in that the onset of vaginal cornification was delayed to a comparable extent. Rather, the data suggest retarded development of ovarian follicles, possibly as a result of pituitary suppression. It is noteworthy that, although delayed, both vaginal cornification and ovulation occurred in spite of continued THC treatment, suggesting the development of drug tolerance.