Patterns of anogenital swelling size and their endocrine correlates during ovulatory cycles and early pregnancy in free-ranging barbary macaques (Macaca sylvanus) of Gibraltar.

In macaques and other cercopithecoid primates, large anogenital swellings (AS) are generally found only in those species in which reproduction is not seasonally restricted. In this respect, the Barbary macaque is unusual because while it shows a marked degree of reproductive seasonality, it also exhibits a striking, exaggerated swelling of the circumanal region and labia. Information on the characteristics of AS in female Barbary macaques is limited in that it is largely based on semiquantitative assessments of swelling size, and there are no data on endocrine parameters associated with AS during ovulatory cycles or early pregnancy. In the present study, we combined quantitative measurements of four swelling size parameters (AS width, height, and depth, and labial width) using a video-imaging technique with fecal estrogen and progestagen determinations in free-ranging females of the Gibraltar Barbary macaque population to 1) characterize the pattern of AS throughout the mating season and early gestation, and 2) examine the relationships among changes in swelling size and endocrine parameters. The patterns of all four swelling parameters correlated significantly with one another, although measures of AS depth and labial width were difficult to obtain. Using the product of AS height and width, the data demonstrate that the occurrence of AS is highly seasonal, with pronounced cyclical changes during the mating season and early pregnancy. Furthermore, the swelling cycles are characterized by progressive size increases from the early to the late follicular phase, in association with an elevated estrogen:progestagen (E:P) ratio, with ovulation occurring during the maximum swelling phase. The results also demonstrated a conspicuous postconception increase in swelling between days 18-30 of gestation. The postconception swellings were on average 80% of the size of that of the conception cycles, and were preceded by a large increase in fecal estrogen levels and the E:P ratio. This is the first study to characterize swelling patterns and their endocrine correlates during ovarian cycles and early pregnancy in naturally reproducing female Barbary macaques. The data provide a solid basis for further studies to explore sociosexual behavioral patterns and the functional significance of AS in this species.

Characterization of urinary and fecal metabolites of testosterone and their measurement for assessing gonadal endocrine function in male nonhuman primates.

The aims of the present study were (i) to provide basic comparative data on the time course, route, and characteristics of excreted [14C]testosterone (T) metabolites in three nonhuman primates: the common marmoset (Callithrix jacchus), the long-tailed macaque (Macaca fascicularis) and the chimpanzee (Pan troglodytes) and (ii) to use this information to help validate the measurement of urinary and fecal testosterone metabolites for assessing androgen status in Anthropoid primates. Radiolabeled 14C-T (10-30 microCi) was injected intravenously into one adult male of each species and the excreta collected over the next 5 days. Peak radioactivity in urine was detected within 2h and accounted for 67% (Mf), 80% (Cj) and 91% (Pt) of the total radioactivity recovered. The time course of excretion of radioactivity in feces showed a higher variation between species (4-26 h to peak values). In all three species, the majority (>90%) of urinary metabolites were excreted as conjugates whereas the proportion of conjugated metabolites in feces was substantially lower and more variable. High pressure liquid chromatography (HPLC) analysis of urinary and fecal extracts revealed multiple peaks of radioactivity in all three individuals, but each with a distinctive pattern. Native T was excreted in only small amounts into the urine, whereas it was virtually absent in the feces of all three individuals. Three C17 group-specific enzymeimmunoassays using antisera against testosterone, 5alpha-androstane-17alpha-ol-3-one and androsterone were evaluated for their ability to discriminate immunoreactive androgen levels between intact males, castrated males and females based on measurements in urine and feces. In the marmoset, all assays (except for T in feces) clearly discriminated between test groups; in the chimpanzee significantly higher levels of androgen immunoreactivity in intact versus castrated males were measured in urine, but not feces. In the macaque, only the 5alpha-androstanolone measurement in feces discriminated between groups. Data on the results of a radiometabolism study using 3H-DHEA (a weak adrenal androgen) in a long-tailed macaque suggested that co-measurement of metabolites derived from T and DHEA in the assays tested might explain the difficulties in discriminating gonadal status in the two Old World primate species. Collectively, the data show that T metabolism in primates is highly complex and that no single method for noninvasive assessment of androgen status can be used for application across species. The importance of a proper validation of the methodology for each species is emphasised.

Comparative aspects of the metabolism and excretion of cortisol in three individual nonhuman primates.

A radiometabolism study is described to provide the first comparative data on the time course, route, and characteristics of excreted [3H]cortisol metabolites in three nonhuman primates: the common marmoset (Callithrix jacchus), the long-tailed macaque (Macacafascicularis), and the chimpanzee (Pan troglodytes). A low dose (40-100 microCi) of 3H-labeled cortisol was administered intravenously to one adult male of each species and the excreta collected over a 5-day period postinjection. The major proportion of radioactivity was excreted in the urine (>80%). Peak radioactivity in urine was recovered within 5.5 h following injection in all three species, while in the feces peak levels of radioactivity were recovered within 26 h postinjection. In all three species, urinary metabolites were primarily excreted as conjugates (61-87%), whereas the percentage of conjugated metabolites in feces was 50% or less. The number and relative abundance of urinary and fecal [3H]cortisol metabolites were determined by reverse-phase high-performance liquid chromatography (HPLC) and immunoreactivity of the radioactivity peaks was assessed by screening HPLC fractions with established cortisol, corticosterone, and 11-oxoetiocholanolone enzyme immunoassays (EIA), the latter being a group-specific assay for measuring 11,17-dioxoandrostanes. HPLC separation of urinary and fecal extracts revealed multiple peaks of radioactivity, several of which were common to all three species. The relative proportion of these peaks, however, differed considerably among species and between urine and feces. HPLC indicated that native cortisol was a major urinary excretory product in the marmoset, while comparatively small amounts were present in the urine of the macaque and chimpanzee. In contrast, in feces, cortisol was only detected in low amounts in the marmoset and was virtually absent in the macaque and chimpanzee. In all three species, one of the major radioactivity peaks showed a retention time comparable to 11-oxoetiocholanolone and high immunoreactivity in the 11-oxoetiocholanolone EIA. The measurement of urinary- and/or fecal-immunoreactive 11,17-dioxoandrostanes is therefore implicated for noninvasive assessment of adrenal function in Old World monkeys, New World monkeys, and great apes.

Efficacy and effects of short- and medium-term contraception in the common marmoset (Callithrix jacchus) using melengestrol acetate implants.

This study examines the effect of melengestrol acetate (MGA) implants on reproductive function and various biochemical parameters, ovarian activity, and uterine morphology in ten female common marmosets implanted for either 6-8 or 19-21 months. Measures of body weight, concentrations of urinary glucose and blood liver enzymes were taken. Ovarian activity was assessed by analysis of urinary progestin levels and ultrasound examinations of the ovaries. Ultrasonography was also used to evaluate uterine morphology. MGA was highly effective in preventing pregnancies in the study animals. No changes in biochemical parameters were found; however, seven females developed a substantial weight gain during the study. Follicular development was not suppressed, as indicated by the presence of antral follicles, luteinized structures, and elevated urinary progestin levels. The uteri of the MGA-treated subjects were moderately enlarged with a thickened endometrium that showed a marked change in structural appearance indicative of hypertrophy and decidualization. After implant removal these changes quickly disappeared and all females ovulated within 3 weeks and conceived within 4 months post-treatment. MGA appears to be an acceptable contraceptive in the marmoset, although non-steroidal methods should be evaluated as possible potential alternatives.

Application of urinary and fecal steroid measurements for monitoring ovarian function and pregnancy in the bonobo (Pan paniscus) and evaluation of perineal swelling patterns in relation to endocrine events.

Estrone conjugates (E1C), pregnanediol glucuronide (PdG), and estriol (E3) in urine, and immunoreactive E1C, E3, pregnanediol (Pd), and progesterone (P4) in feces were determined along with records of perineal sex skin swelling throughout 7 nonconception cycles and 3 full-term pregnancies of 4 adult female bonobos (Pan paniscus). A typical preovulatory urinary E1C surge and postovulatory increase in urinary PdG were seen during the menstrual cycles. Fecal progestin levels were significantly correlated with those of PdG in urine in all cycles, while E1C measurements in feces were significantly correlated with those in urine in only 3 cycles. On the basis of hormone profiles, a variable follicular phase of 17-40 days and a relatively constant luteal phase of 11-15 days was found, resulting in cycle lengths of 31-51 days. All urinary and fecal hormones were markedly elevated during pregnancy. Measurement of E1C in both urine and feces was most useful for early pregnancy diagnosis, while E3 was of value in confirming pregnancy and assessing fetal viability. The period of perineal swelling during the cycle comprised on average 66.3% of cycle length, half of which was associated with a phase of maximum tumescence. Ovulation usually occurred within the maximum swelling phase, but timing of ovulation within this period was highly variable and was more closely associated with the end rather than the onset of maximum tumescence. The data presented here are of great practical value in the captive breeding management of bonobos and offer new opportunities for investigating basic questions of bonobo reproductive biology both in captivity and in the wild.

Synthesis and cell surface display of class II determinants by long-term propagated rat T line cells.

We have investigated the capacity of the encephalitogenic BS rat T cell line bs 83 and its variant clone bs 83.III.C6 to synthesize and express RT1.B-specific class II molecule subsets defined by monoclonal antibodies (mAb) MRC-OX6 and MRC-OX3. Earlier studies had indicated that mAb MRC-OX6 recognizes three distinct molecular species: an immature oligomeric polypeptide chain complex comprised of the polymorphic subunits alpha, beta and the invariant proteins of the gamma group; a biosynthetic intermediate composed of post-translationally modified alpha, beta and gamma chain (denoted p35) and a fully glycosylated alpha, beta two-chain complex derived from the plasma membrane. MRC-OX3 was shown to recognize a serologically distinct alpha, beta two-chain complex that coexists with the MRC-OX6-specific heterodimer at the cell surface. Here we show that premutant bs 83 cells were unable to synthesize class II molecules of either set. In contrast endogeneous synthesis by mutant cells of MRC-OX6-specific molecules was demonstrated. Unlike control spleen cells variant cells failed to synthesize the mature MRC-OX3-reactive class II subset. Instead a three-polypeptide chain complex comprised of the terminally glycosylated subunits alpha, beta and invariant chain p35 was present at the cell surface. This complex appears to represent the preserved biosynthetic intermediate that failed to release invariant chain p35 upon its transit into the plasma membrane. These latter observations support our notion of gamma chain-induced epitope diversification during post-translational maturation of RT1.B-specific class II molecules.