Growth and reproductive parameters of bonnet monkey (Macaca radiata)

The present paper summarizes some of the important biological and physiological data recorded over a 30-year period on the biology of bonnet monkeys in captivity. Data on sexual maturity, menstrual cyclicity, general behaviour, endocrine profile, reproductive physiology, gestation, parturition, postpartum amenorrhoea in the female, and sexual maturity, hormone profile, and seasonal variation in sperm count of the male monkeys are presented. In addition to the biological values, weights of selected organs, vertebral and dental pattern are also presented. Menarche occurred at an age of 36±4 months and the first conception in the colony occurred at an age of 54±4 months. The average menstrual cycle length was 28±4.3 days. Majority of monkeys did not cycle regularly during March–June during which the temperature reached a peak. The pregnancy index of the colony was 80% with controlled breeding. The gestation period was 166±5 days with 6–7 months postpartum amenorrhoea. Males attained sexual maturity by the age of 6–7 years and exhibited the characteristic nocturnal surge of serum testosterone at this age and sperm concentration ranged from 116–799 millions/ejaculate.     

Studies on ovarian quiescence in the lactating bonnet monkey (Macaca radiata)

Lactating bonnet monkeys were used as a model to understand the mechanism of ovarian quiescence during lactation. The ovary of the bonnet monkey in the 3rd month of lactation responds well to exogenous pregnant mare serum gonadotropin stimulation with serum estrogen values reaching maximal levels by day 3 of the gonadotropin injection. The adminstration of ovine prolactin to such monkeys significantly inhibited the ovarian responsiveness to exogenous gonadotropin. The responsiveness of the pituitary of the lactating monkey (in the 3rd month of lactation) to luteinizing hormone releasing hormone injection was suppressed and supplementation with exogenous prolactin further accentuating this effect. The relative ability of chlorpromazine given intravenously/intramuscularly/intranasally to enhance endogenous prolactin levels was assessed. During the first 5 months of lactation when the basal prolactin levels were high, the luteinizing hormone levels were low. As the suckling stimulus reduces and prolactin levels fall, luteinizing hormone levels increase, the first post-parturient mensus occurring by 218 ± 4 days. This event was postponed by 3 months on increasing endogenous prolactin levels by administering chlorpromazine (250 μg/day by intranasal mode) over a 5 day period every month starting from the 3rd month of lactation.     

Estrogen modulation of riboflavin carrier protein in the bonnet monkey (Macaca radiata)

Circulatory concentrations of riboflavin carrier protein (RCP) were quantitated in bonnet macaques by employing a heterologous radioimmunoassay involving 125I-labelled chicken RCP and its antiserum. The levels of monkey RCP in the serum seem to be governed by the estrogenic status of the animals. An increase in concentration of serum estradiol in the adult females during the menstrual cycle and early pregnancy could be correlated with enhanced serum RCP levels. Estadiol-17 beta administered to both immature female and male monkeys, specifically brought about elevated levels of RCP with a slower time course of response in males than in females. These results could be a reflection of a more rapid decline of both circulatory estrogen and RCP concentrations in male serum. Repeated administration of estradiol-17 beta to male animals led to prolonged elevated levels of RCP following estrogen administration. Thus, it would appear that the evolutionary conservation of RCPs from the aves to the primates encompasses not only their physicochemical similarities but also extends to the estrogenic modulation of their elaboration.     

Estradiol levels near the time of ovulation in the bonnet monkey (Macaca radiata)

Plasma estradiol-17beta and total progestins were determined to delineate the relationship between preovulatory estradiol-17beta peak and ovulation in the bonnel monkey (Macaca radiata). 6 monkeys were studied for 15 menstrual cycles. In subsequent cycles, serial laparotomy was performed in 5 of the 6 monkeys to correlate ovarian morphology to plasma estradiol-17beta. In 11 of the 15 cycles, estradiol-17beta peaks were 3- to 7-fold above baseline levels near the time of expected ovulation (Cycle Days 7-12). Plasma progestin rose significantly from follicular phase levels of .5 ng/ml to 2.6 ng/ml the day of the estradiol-17beta peak with peak levels of 4.5 ng/ml on the following day. Ovarian morphology in 4 of the 5 observed by laparotomy demonstrated ovulation within 48 hours following an estradiol-17beta peak of approximately 300 pb/ml.     

Antifertility effect of tamoxifen as tested in the female bonnet monkey (Macaca radiata)

The administration of a potent antiestrogen, tamoxifen at a dose of 3 mg/kg body weight/day orally post-coitally to cycling mated bonnet monkeys(Macaca radiata) from days 18–30 of cycle resulted in inhibition of establishment of pregnancy in 9 out of 10 monkeys. Tamoxifen effect was not due to interference with luteal function. The effect was specific to tamoxifen as exogenously administered progesterone could not reverse it. In addition to suggesting a role for estrogen in maintenance of early pregnancy in the primate the present study could be a prelude to the development of an effective post-ovulatory approach for regulation of fertility in the human female     

Diurnal variations of serum testosterone levels in the male bonnet monkey (Macaca radiata)

Diurnal variations in serum testosterone levels were studied in six adult maleMacaca radiata. Though the testosterone levels fluctuated considerably during the 24-hr period, a distinctive diurnal pattern characterized by lower concentrations during day hours and higher concentrations during night hours was observed.     

Temporomandibular malformations in the bonnet monkey (Macaca radiata) fetus following maternal ingestion of thalidomide

Structural abnormalities of the temporomandibular region were seen and described in the fetal bonnet monkey (Macaca radiata) after maternal ingestion of thalidomide. A single oral dose of 10 mg/kg of thalidomide was given on day 24, 25, 26, 27, 28 or 29 or 30 mg/kg on day 25 or 28 of gestation to 25 pregnant monkeys (day of mating = 0). The skeletons, processed and stained with alizarin red S, were examined for gross changes in configuration of craniofacial bones. Temporomandibular defects, in varying severity, were seen in four 70-day-old and seven 100-day-old fetuses associated with a single treatment between day 24 and day 28. The highest risk appeared to occur following treatment on day 25 (4/5) and the lowest on day 28 (2/8). Characteristic expression of the defects included 1) temporo-zygomatic hyperplasia involving the articular tubercle and postglenoid process and 2) mandibular hypoplasia, primarily of the condyles and the angular and coronoid processes. Micrognathia, apertognathia, and partial extraarticular ankylosis were seen. Although the sensitivity periods of the temporomandibular and ear regions seem to overlap and the embryonic origins are similar, the defects of each region appeared to be independent. Although the ear demonstrated a definite temporal relationship characterized by a decrease in severity in the expression of the ear malformations with respect to increased gestational age at the time of thalidomide insult, the temporomandibular region failed to exhibit a specific pattern. Although the temporo-zygomatic hyperplasia was dissimilar to any reported human condition, the mandibular changes were similar.     

Social interactions of juvenile female bonnet monkeys,Macaca radiata

This study reports observations on juvenile female bonnet macaques (Macaca radiata) in a field cage environment. Interaction and proximity data collected on different age classes of juvenile females indicate distinct changes in social behavior from the time a female is weaned until her first pregnancy. Behavioral development includes an increase in solitary behavior, an early disinterest in younger animals, and a surprisingly low interaction rate with relatives. Important aspects of ontogeny are lost in analyses that treat all juvenile females as a single age-class. This research was supported partially by US PHS Grant No. RR00169.     

Correlation of seasonal changes in sperm output with endocrinological changes in the adult male bonnet monkey,Macaca radiata

We have examined the monthly variations in sperm output and attempted to correlate the profiles of endocrine hormones secreted with the sperm counts throughout the year in the adult male bonnet monkey. As previously reported, there was a distinct spurt in sperm output beginning September through December months. A concomitant increase in serum testosterone and prolactin concentrations were also noted during September through November (mid and post-monsoon season). Although there was a marked increase in gonadotropin releasing hormone stimulated testosterone secretion, the peak testosterone concentrations post gonadotropin releasing hormone injection did not vary significantly (P > 0.05) throughout the year. Basal serum follicle stimulating hormone concentrations did not vary significantly (P > 0.05) during April to June months compared to September-November months. Serum inhibin concentration remained unaltered throughout the year, except in the month of March. The results of this study provide evidence for annual rhythms in prolactin and testosterone secretion and a distinct seasonality in the sperm output of the adult male bonnet monkey, but the pituitary responsiveness to exogenous gonadotropin releasing hormone remains unaltered throughout the year. Because of the existence of seasonality as noted in the present study, future studies which utilize the adult male bonnet monkey as an experimental model need to take into consideration the seasonal effects on reproductive function in this species.     

Granulosa theca cell tumor with luteoma in the ovary of a bonnet monkey (Macaca radiata)

A mass was identified on the left caudal region of the abdomen in a 13-year-old bonnet monkey (Macaca radiata). The mass was excised and diagnosed as granulosa theca cell tumor accompanied with luteoma based on the microscopic findings. Morphologically it appeared pink, round, firm multilobulated measured approximately 5 x 3 x 2.5 cm in dimension. Histologically the luteoma composed of polyhedral cells with pale strained vacuolated cytoplasm, centrally located nuclei with distinct cytoplasmic borders. Granulosa theca cell tumor appeared as densely packed spindle shaped fusiform cells arranged in interlacing bundles and whorled pattern with neoplastic cells appearing irregular shaped solid sheets. The concomitant development of granulosa theca cell tumor with luteoma in a single ovary is very rare and is the first reported case in a bonnet macaque to our knowledge.     

Effect of FSH deprivation at specific times on follicular maturation in the bonnet monkey (Macaca radiata)

Cyclic monkeys were deprived of FSH for specific periods on different days of the follicular phase by injecting them with minimal doses of an FSH antiserum characterized for specificity and bioneutralizing ability. The effect of the antiserum on follicular maturation was assessed by determining (a) serum oestrogen concentrations through the midcycle period, (b) serum progesterone concentrations as an index of ovulation and luteal function, (c) laparoscopic examination of the surface of the ovary when necessary, and (d) overall cycle length. While antiserum injection on Day 5 of the cycle caused delay in the oestrogen surge from Days 9 to 11, injection on Day 6 led to the occurrence of two oestrogen surges, on Days 9 and 14. Laparoscopic examination showed that the earlier follicle had disappeared and a new follicle had appeared by Day 14. Antiserum injection on Day 7 of the cycle arrested further growth of the maturing follicle, but a new follicle appeared 9 days later, as indicated by a surge of oestrogen on Day 16. Injection of antiserum beyond Day 7 had no effect on follicular development, ovulation and luteal function. These observations suggest that the mature follicle becomes relatively independent of FSH support about 48 h before ovulation and this event could be a marker for follicular dominance.     

Serum testosterone levels during the menstrual cycle and early pregnancy in the bonnet monkey (Macaca radiata)

Serum testosterone concentrations have been determined during the menstrual cycle and early pregnancy in the bonnet monkey, Macace rediata. During the cycle, there is an increase around the time of ovulation and a secondary peak in the late luteal phase. In pregnancy, there is a distinct peak around 23-25 days, a period which corresponds to the peak of chorionic gonadotropin reported by Atkinson et al. (1975) in Rhesus monkeys. Administration of exogenous hCG causes a significant rise in the serum testosterone level in cycling monkeys.     

Refolding, structural transition and spermatozoa-binding of recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C expressed in Escherichia coli.

An internal cDNA fragment (978 bp) corresponding to bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C (bmZPC), excluding the N-terminal signal sequence and the C-terminal transmembrane-like domain, was cloned in pQE-30 vector and the protein expressed as inclusion bodies in Escherichia coli. Recombinant bmZPC (r-bmZPC) was solubilized from purified inclusion bodies in the absence of a high concentration of chaotropic agents and was subsequently refolded. Use of a low concentration of urea (2 M) during solubilization of r-bmZPC helped to minimize the extent of protein aggregation during refolding of the recombinant protein, and retain the existing native-like secondary structure that was essential for proper folding. Purified r-bmZPC appeared as a dominant band of 43 kDa on SDS/PAGE and Western blot. Although it lacked carbohydrate moieties, the purified and refolded r-bmZPC bound to the head region of bonnet monkey spermatozoa, confirming the existence of a native-like conformation. CD revealed a maximum at 200 nm and a single broad minimum extending from 209 to 216 nm, indicating the presence of both alpha-helical and beta-sheet conformations in the refolded r-bmZPC. Two different phases of transition were observed by urea-gradient electrophoresis, suggesting the existence of multiple intermediate stages during the unfolding of r-bmZPC. The availability of refolded r-bmZPC will help in elucidating its role during the complex cascade of events during fertilization.