The role of FSH and diurnal testosterone rhythms in specific germ cell transformations during spermatogenesis were investigated using DNA flow cytometry and morphometry of the seminiferous epithelium of the adult male bonnet monkey (Macaca radiata), the endogenous hormone levels of which were altered by two different protocols. (1) Active immunization of five monkeys for 290 days using ovine FSH adsorbed on Alhydrogel resulted in the neutralization of endogenous FSH, leaving the LH and diurnal testosterone rhythms normal. (2) Desensitization of the pituitary gonadotrophs of ten monkeys by chronically infusing gonadotrophin-releasing hormone analogue, buserelin (50 micrograms/day release rate), via an Alzet pump implant (s.c.) led to a 60-80% reduction in LH and FSH as well as total abolition of testosterone rhythms. The basal testosterone level (3.3 +/- 2.0 micrograms/l), however, was maintained in this group by way of an s.c. testosterone silicone elastomer implant. Both of the treatments caused significant (P < 0.01) nearly identical reduction in testicular biopsy scores, mitotic indices and daily sperm production rates compared with respective controls. The germ cell DNA flow cytometric profiles of the two treatment groups, however, were fundamentally different from each other. The pituitary-desensitized group exhibited a significant (P < 0.001) increase in 2C (spermatogonial) and decrease in 1C (round spermatid) populations while S-phase (preleptotene spermatocytes) and 4C (primary spermatocytes) populations were normal, indicating an arrest in meiosis caused presumably by the lack of increment in nocturnal serum testosterone. In contrast, in the FSH-immunized group, at day 80 when the FSH deprivation was total, the primary block appeared to be at the conversion of spermatogonia (2C) to cells in S-phase and primary spermatocytes (4C reduced by > 90%). In addition, at this time, although the round spermatid (1C) population was reduced by 65% (P < 0.01) the elongate spermatid (HC) population showed an increase of 52% (P < 0.05). This, taken together with the fact that sperm output in the ejaculate is reduced by 80%, suggests a blockade in spermiogenesis and spermiation. Administration of booster injections of oFSH at time-points at which the antibody titre was markedly low (at days 84 and 180) resulted in a transient resurgence in spermatogenesis (at day 180 and 228), and this again was blocked by day 290 when the FSH antibody titre increased.(ABSTRACT TRUNCATED AT 400 WORDS)

Testicular germ cell populations of biopsies from 32 male bonnet monkeys in 5 different age groups were quantitated in a flow cytometer after labelling of germ cell DNA with the specific fluorochrome, 4,6-diamidino phenyl indole. The 5 quantifiable populations were spermatogonia (2C), preleptotene spermatocytes (S phase), primary spermatocytes (4C), round spermatids (1C) and elongate spermatids (HC). The seminiferous tubules of immature 3-4-year-old monkey had only Sertoli cells and spermatogonia (2C). At 5-6 years, germ cells in S-phase (9.5%), 4C (11.1%), 1C (41.8%) and HC (17.1%) stages of maturation appeared for the first time but at 7-8 years of age and beyond all cell types except HC decreased while 1C remained relatively constant. Histometric analysis correlated well with the flow-cytometric data. The decrease in cells of 2C, S-phase and 4C stages was associated with an increase in mitotic index, signifying acceleration in the kinetics of germ cell transformation into subsequent cell types. The total turnover in cell transformation (1C:2C) was significantly (P less than 0.01) increased at and beyond 7-8 years. Maximum transition from 2C to 4C occurred at 5-6 years (4C:2C ratio 0.8 at 5-6 years and 0.6 at 7-8 years). The ratio HC:1C (kinetics of cell transformation during spermiogenesis) attained near total efficiency only by 10 years of age (1.08 at 10-14 years; 0.9 at 18-20 years). Also, the cell associations within the seminiferous tubules of monkeys greater than or equal to 10 years of age were better defined than those of younger animals. The changes in germ cell ratios correlated well with alterations in testicular volume, sperm numbers in the ejaculate and surges of testosterone and increments in FSH in the serum, characteristic of development of sexual maturity. It is apparent from this study that DNA flow cytometry of testicular germ cell populations reveals subtle changes in spermatogenic status of bonnet monkeys with a high degree of sensitivity.

Administration of Tamoxifen via the Alzet pump at a rate of 50 pg hr-' for 90 days in the adult male bonnet monkeys Macaca radiata had no effect on the serum testosterone concentration determined at 10 AM and 10 PM as well as total sperm count determined at 15-day intervals over a period of 260 days. However, a significant reduction in sperm motility was observed beyond 90 days up until the 225th day. Breeding studies conducted from day 90 to 260 revealed that these males were infertile.

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A simple method is described to collect cervical smears and to have a clear view of cervix for small breed of monkeys. This method was found useful to collect adequate cytology smears with good preservation of cellular morphology.