Administration of human luteinizing hormone (hLH) to macaques after follicular development: Further titration of LH surge requirements for ovulatory changes in primate follicles

After stimulation of multiple follicular development, endogenous LH surges elicited by GnRH or GnRH agonist were of insufficient duration (4-14 h) to evoke oocyte maturation and luteinization in this species. In this study, periovulatory LH surge requirements were further titrated using hLH as the ovulatory stimulus. Beginning at menses, rhesus monkeys were treated with human gonadotropins for 9 days to stimulate follicular growth. To induce ovulatory maturation on day 10, animals received: 1) hCG (1000 IU, im; n = 8); 2) highly-purified, urinary hLH (2542 IU, im; n = 4); or 3) hLH (2542 IU, im) followed by three injections of hLH (200 IU, im) at 8-h intervals (0800, 1600, 2400 h) daily during the luteal phase until menses (n = 3). Oocytes and luteinizing granulosa cells were obtained via follicle aspiration 27 h after the initial hLH or hCG injection. Estradiol and progesterone levels were measured in daily serum samples by RIA. Bioactive LH levels were determined at selected intervals within 36 h of the hLH ovulatory stimulus. Nuclear maturity of oocytes was evaluated as an indicator for reinitiation of meiosis. Luteinizing granulosa cells were processed for indirect immunocytochemistry using a monoclonal antibody to human progesterone receptor. In vitro progesterone production by luteinizing granulosa cells over 24 h was also assessed in the absence and presence of hCG. In all groups, serum estradiol rose to similar peak levels on day 10. After hLH, bioactive LH levels peaked (1262 ± 79 ng/mL; mean ± SEM) by 2-6 h, declined thereafter but remained above surge levels (100 ng/mL) for 18-24 h. Within 24 h of hLH injection, serum progesterone increased to 13 ± 3 nmol/L, but returned to baseline in 1-6 days. In contrast, higher levels of progesterone were observed after hCG (114 ± 51 nmol/L) and during luteal phase treatment with hLH (137 ± 25 nmol/L) and the luteal phase was longer (11.5 ± 0.4 and 14.3 ± 0.7 days, respectively). Of the total cohort of oocytes aspirated, the proportion of oocytes resuming meiotic maturation (metaphase I plus metaphase II) was similar after hCG (76%) and hLH (74%). However, the proportion of oocytes maturing to metaphase II tended to be less (P = 0.08) after hLH (13%) than hCG (22%). Fertilization rates were similar between the two groups. Progesterone receptor was detected in nuclei of luteinizing granulosa cells from all animals receiving hCG, but only in some given hLH. Basal progesterone production by luteinizing granulosa cells was greater after hCG than hLH treatment (1695 ± 572 and 563 ± 216 nmol/L, respectively). The addition of hCG in vitro enhanced progesterone secretion by cells from both hCG- (2798 ± 598 nmol/L) and hLH-treated animals (1829 ± 569 nmol/L). Thus, an interval of LH exposure (18-24 h) half of that of the spontaneous LH surge during a normal menstrual cycle induced early ovulatory changes in primate follicles, but failed to sustain corpus luteum function. “Pulsatile” exposure to hLH after the truncated LH surge restored luteal function and life span to that reminiscent of the normal cycle.