Existence of multiple forms of follicle-stimulating hormone within the anterior pituitaries of cynomolgus monkeys.

Ovariectomized cynomolgus monkeys were treated with physiological levels of estradiol and progesterone. A reduction in serum levels of FSH was observed after steroid exposure. Anterior pituitary homogenates were prepared from monkeys after 0, 12, 24, or 36 h of exposure to estradiol and progesterone and quantitated for FSH activity by radioreceptor assay (RRA) and RIA. Pituitary FSH activity (expressed as RRA/RIA) increased with duration of exposure to steroids. Forms of FSH within these pituitaries were separated by the column isoelectric focusing technique, chromatofocusing. All pituitary homogenates tested contained FSH isohormones that eluted at similar isoelectric points. Each FSH isohormone exhibited a mol wt similar to that of a purified FSH standard, but differed in ability to displace labeled FSH from a biological receptor preparation. FSH forms with basic isoelectric points exhibited greater RRA/RIA values than forms with more acidic isoelectric points. The relative proportion of the more basic FSH forms increased within pituitary tissue with duration of exposure to steroids. All FSH forms were secreted by pituitary cells in culture. The biochemical basis for the microheterogeneity appears to be the degree of sialic acid incorporation into the FSH molecule. The results of these studies demonstrate that the cynomolgus monkey pituitary responds to the surrounding hormonal milieu by altering the relative proportions of FSH forms present within that gland.

Assessment LOPU-IVF in Japanese sika deer (Cervus nippon nippon) and application to Vietnamese sika deer (Cervus nippon pseudaxis) a related subspecies threatened with extinction.

In mammals, recovery of oocytes by laparoscopic ovum pick-up (LOPU) coupled with in vitro production (IVP) of embryos represents a promising strategy for both amplification and genetic management of sparse animals from captive endangered wild species. As integrated technique developed mainly for domestic livestock, LOPU-IVP requires several studies to set up protocols for follicular stimulation or optimization of IVP before envisaging successful transposition to wild species. In deer, many endangered subspecies would be potentially concerned by applying such an approach using common subspecies for protocols optimization. The aim of the present study was to assess efficiency of follicle stimulation using ovine FSH (oFSH) for recovery of oocytes by LOPU in common sika deer (Cervus nippon nippon) before transposition of an optimized methodology for IVP of embryos from endangered Vietnamese sika deer hinds (Cervus nippon pseudaxis). In common sika deer, two doses of oFSH (0.25 and 0.5 U) and two frequencies of administration (12 and 24 h) were compared by monitoring of subsequent ovarian response, quality of oocytes recovered by LOPU, and in vitro developmental competence. In a first experiment, the dose of oFSH administered did not significantly affect the total number of follicles aspirated per hind per session (8.6 ± 1.0 vs. 8.2 ± 1.6 with 0.5 vs. 0.25 U oFSH, respectively; not significant). In a second experiment, frequency of 0.25 U oFSH administration did not affect ovarian response. Efficiency of IVP determined on blastocysts rates after in vitro maturation, fertilization, and development in oviduct epithelial cells coculture was increased when FSH was administered at 12-h intervals. Immune response after several follicular stimulations was detected against exogenous oFSH in plasma from the majority of sika deer hinds but was not associated with decreased ovarian response. When 0.25 U oFSH was administered at 12-h intervals to Vietnamese sika deer (N = 4), good quality cumulus oocyte complexes with complete and compact cumulus investments were recovered allowing a high cleavage rate after in vitro maturation and fertilization. Development to the blastocyst stage occurred in a high proportion (30% of oocytes) after coculture with ovine epithelial cells allowing cryobanking of transferable embryos from Vietnamese sika deer. These results confirm that LOPU-IVF after ovarian stimulation with oFSH may be a successful tool for cryobanking transferable embryos from endangered sika deer subspecies.

Alpha and beta subunit immunoreactivity of reference preparations used in the radioimmunoassay of follicle stimulating and luteinizing hormones in serum.

The 2nd IRP-HMG, the 1st IS, and LER 907 have been widely used in standard FSH and LH RIA systems to measure circulating levels of FSH and LH in human serum. In these systems, the dose-response curves elicited by the preparations were parallel to each other and to that of serum samples tested. When these materials were studied in the homologous alpha and beta-hLH RIA systems, the three hormones and a serum sample used as internal control in all assays, displayed a higher degree of contamination with alpha than with beta-hLH subunits. On weight basis, the alpha/beta ratios were 18.0, 18.7, 6.7, and 13.0, for the 2nd IRP-HMG, 1st IS, LER 907 and serum, respectively. In the homologous alpha and beta hfSH RIA's, the highest contamination with alpha-hfSH was displayed by (alpha/beta ratio:10), the alpha/beta ratios for the 2nd IRP-HMG and 1st IS were 2.2 and 1.1, respectively. The serum sample contained about 6.2 ng equivalent of alpha-hFSH/ml; beta-hFSH was not detected.