Caprine luteinizing hormone isoforms during the follicular phase and anestrus.

The relative proportion of the circulating luteinizing hormone isoforms in goats during follicular phase (pre-ovulatory peak; F) and anestrus (A) was investigated. Estrus was synchronized in six goats with a prostaglandin analogue. After estrus was detected, blood samples were taken at 1 h intervals for 24 h. Four anestrous goats received 100 microg i.v. of GnRH and blood samples were collected every 15 min for 5 h. Samples with the greatest LH concentration in follicular phase and after GnRH administration (anestrus) were analyzed by chromatofocusing and eluted with a pH gradient from 10.5 to 3.5. For quantification purposes eluted LH was grouped into basic (pH> or =7.5), neutral (pH 7.4-6.5) and acidic isoforms (pH< or =6.4) as well as by pH unit. In both physiological conditions (PC), basic and acidic isoforms were greater than the neutral. With this grouping criteria, there was an interaction between PC and pH group, with the proportion of neutral isoforms being greater (p<0.05) in A (12.0+/-0.8%) as compared with F (5+/-2%). Analysis by pH unit showed a very basic group of eluted isoforms (pH> or =10), which amounted to a percentage of 6.0+/-0.4% of the total observed during A, and 3+/-1% during F (p<0.05). Predominant isoforms in A eluted in the pH range 9.99-9.0 (42+/-3%) as compared to 7+/-3% (p<0.01) in that pH range in F. In contrast, the predominant isoforms in F eluted in the pH range 8.99-8.0, representing 55+/-8%, while in A the proportion was 11+/-2% (p<0.01). Isoforms eluted at the pH range 7.9-7 represented a significantly greater proportion during A (5.0+/-0.6%) as compared with F (3+/-1%). This is the first report on goat LH circulating isoforms. During A the LH isoforms secreted by the pituitary are more basic than during F.

Artificial insemination in black-handed spider monkey (Ateles geoffroyi).

Artificial insemination (AI) was performed in spider monkeys; these primates are vulnerable to extinction and usually do not reproduce spontaneously in captivity. Uterine cycles were followed by daily assessment of vaginal cytology, and corroborated a posteriori by concentrations of 17-beta estradiol and progesterone, measured by radioimmunoassay (RIA), in fecal samples collected once daily. Five females between 13 to 27 years old were inseminated intravaginally (with fresh semen) twice each during the periovulatory phase (Days 9-12 of the menstrual cycle; Day 0, first day of menstrual bleeding), from September to the first 3 weeks of November (most fertile months). Transcervical AI was not useful in this primate because the liquid portion of the semen completely solidified instead of liquefying as in other primates. Pregnancies were apparently achieved in 5 of 14 attempts. One female became pregnant after the first round of inseminations, delivered a healthy infant, was inseminated and got pregnant again (subsequently aborted). One female aborted, apparently due to an intramural uterine leiomyoma. Another two females stopped menstruating for a few months, then restarted menstruating (these females may have been pregnant and aborted). In conclusion, in spider monkeys: (1) captivity-induced stress did not inhibit reproduction; (2) fecal steroid hormones were useful to assess cyclicity; (3) the semen coagulum, which apparently is a tightly packed and large reservoir of spermatozoa, must not be discarded but used in AI; (4) old female spider monkeys did not have cessation of reproductive function.

Seasonal variations of fecal progesterone and 17beta-estradiol in captive female black-handed spider monkeys (Ateles geoffroyi).

A number of studies in free-ranging and captive spider monkeys (Ateles spp.) have shown that this genus is able to reproduce throughout the entire year. Nonetheless, it is still controversial whether births, and therefore conceptions, tend to be more frequent during certain seasons. In the present study, we monitored changes in fecal 17beta-estradiol and progesterone for approximately 1 years in five female black-handed spider monkeys (Ateles geoffroyi) kept in captivity in Mexico City. The objective was to determine whether hormone concentrations and menstrual cycles of summer and autumn accounted for a greater chance of conception than those of winter and spring, consistent with birth patterns previously reported. We collected fecal samples from the five monkeys almost daily for 1 year (March 2004 to February 2005) and used radioimmunoassay of fecal extracts to determine concentrations of 17beta-estradiol and progesterone. Concurrently, menstrual cycle phases were determined by cytological evaluation of vaginal swabs. Periovulatory 17beta-estradiol concentrations were significantly higher in autumn than in winter, spring or summer. Moreover, as evidenced by progesterone peaks, most of the summer-autumn menstrual cycles were ovulatory; in contrast, most of the winter and spring cycles were anovulatory. In conclusion, our data supported the notion that, although not a strictly seasonal reproducer, the black-handed spider monkey is more likely to conceive at the end of the rainy season and throughout autumn.

Ovulation and luteal characteristics following removal of the ovine corpus luteum (lutectomy) at four times during the oestrous cycle.

The life span of the corpus luteum (CL) may depend on follicular development. To provide evidence relating to this hypothesis, each of 32 ewes was randomly assigned to have its CL removed on day 2, 3, 4 or 10 after oestrus. Twenty ewes were treated with 1000 IU of human chorionic gonadotrophin (hCG) 36 h after CL removal to induce ovulation; the other 12 ewes were not treated with hCG. Blood samples were collected daily to monitor the ovulatory response and the characteristics of the next cycle at the first sign of oestrus and up to day 21 after surgery or hCG administration. Every animal ovulated within 7 days of hCG administration, regardless of when its CL had been removed. It was concluded that the follicles found in the ovary as early as the second day after oestrus respond to endogenous or exogenous ovulatory stimuli affecting the life span of resulting CL.

Increased cleavage and blastocyst rate in ewes treated with bovine somatotropin 5 days before the end of progestin-based estrous synchronization.

Treatment with bovine somatotropin (bST) during estrous synchronization increased fertility and prolificacy in sheep. In the present study, a single dose of bST 5 days before the end of progestin treatment improved cleavage and embryo development. Stage of estrous cycle was synchronized in ewes (n=32) with progestin and superovulation was induced by use of FSH. Five days before the end of progestin treatment, ewes were randomly assigned to two groups: bST group (n=16) received a depot injection of 125 mg of bST sc (Lactotropina, Elanco, México) and the control group (n=16) received saline solution. Estrous was detected with rams fitted with an apron every 2 h and estrous sheep were mated every 8 h whilst in estrous. Embryos were recovered on Day 7 post mating, assessed microscopically and fixed in 4% paraformaldehyde. Cell number in blastocysts was counted after Hoechst 33342 staining. Plasma concentrations of IGF-I, insulin and progesterone were determined in eight sheep per group from the day of bST treatment to the day of embryo recovery. Cleavage rate, percentage of transferable embryos (transferable embryos/cleaved) and percentage of embryos reaching the blastocyst stage (blastocyst/cleaved) were compared between groups by logistic regression. IGF-I, insulin and progesterone plasma concentrations were analyzed by ANOVA for repeated measurements and cell number by ANOVA. Cleavage rate was greater (P<0.01) in bST treatment group (86%) than in the control group (62%). Similarly, the proportion of embryos reaching the blastocyst stage (bST=68.7 vs control=42.5) and the number of cells per blastocyst (bST group 91.8±5.5 compared to control group 75±6) were greater (P<0.01) in the bST-treated sheep. Plasma concentrations of IGF-I and insulin were greater (P<0.01) in the bST-treated group. No changes were observed in progesterone concentrations (P=0.5). It is concluded that bST injection 5 days before progestin removal increases cleavage rate and the proportion of embryos that reach the blastocyst stage. These effects are associated with an increase in IGF-I and insulin concentrations.

The menstrual cycle of the spider monkey (Ateles geoffroyi).

The ovarian cycles of four adult female spider monkeys (Ateles geoffroyi) were followed daily throughout 30 days by means of vaginal swabs and blood samplings. Cytological analyses of the vaginal swabs and radioimmunoassay determination of the daily levels of estradiol-17 beta (E2) and progesterone (P4) wer done in order to classify the kind of ovarian cycle of this species. Our results show that Ateles geoffroyi females display menstrual cycles of about 24 days on average. By comparison with the well-known menstrual cycles of women, apes, and Old World monkeys, the four distinctive cytological phases (bleeding, follicular, periovulatory, and luteal) could be recognized; mid-cycle E2 peaks followed by mid-luteal increases of the same hormone were present in all four females. P4 levels were higher after the E2 peak, although both hormones were present throughout the cycles. Also, age-dependent features, hormone profiles, and changes in menstrual phases lengths were detected.