Assessment of ovarian response to gonadotropins

Procedure: To examine ovarian response to gonadotropins, 70 days old mature rats from respective control (n=10), and experimental groups (n=12) were induced to superovulate by subcutaneous (sc) injection of 40 IU PMSG at 0 h and 25 IU hCG at 52 h. After 15-20h of hCG injection, the rats were sacrificed by cervical dislocation. The peritoneum was cut open and the fallopian tube along with a part of uterus was carefully excised out. The tube was then cleared off adhering fat and washed in preequilibrated PBS at 370C. The separated oviduct along with the small part of uterus was placed under the stereo-microscope and the fimbrial end was located out. Then with the help of a blunt 30gauge needle fitted to a tuberculin syringe, the tube was flushed from the ovarian end with pre- equilibrated PBS at 370C. The tubal flushings were collected in a grooved slide and 0.1% hyaluronidase was added to the medium to disperse the cumulus cells surrounding the oocyte [Bandopadhyay et al., 2003]. Presence or absence of oocytes and their numbers were recorded.

Bandyopadhyay S, Chakrabarti J, Banerjee S, Pal AK, Goswami SK, Chakravarty BN, Kabir SN 2003; Galactose toxicity in the rat as a model for premature ovarian failure: an experimental approach readdressed Hum Reprod 18 :2031-2038.