Isolation, identification and characterization of secretory proteins of IVMFC embryos and blood circulation of estrus and early pregnant goat.

The aim of the present study was to isolate, identify and characterize the secretory proteins of IVM oocytes and IVMFC embryos to evaluate its immunogenecity. and identify of such proteins if any, in blood circulation of estrus and early pregnant goats. Oocytes were matured in TCM-199 with 1 microg/ml, estradiol-17beta; 0.5 microg/ml, FSH; 100 IU/ml, LH and 10% FCS on granulosa cell monolayer. After 18 hr of maturation, oocytes were further cultured in maturation medium containing 3 mg/ml polyvinyl alcohol (PVA) without serum and BSA for 12 hr and medium was collected. The IVF embryos of 4-8 cell stage were cultured in medium containing PVA without serum and BSA. Embryo culture medium was collected after 24 hr of culture and was pooled. The proteins were analyzed on SDS-PAGE (12.5%). Four secretory proteins of oocytes with approximately molecular weight of 45, 55, 65 and 95 kDa and three secretory proteins of embryos 45, 55 and 65 kDa were obtained on SDS-PAGE in silver staining. The protein profile of midluteal, estrus and early pregnant goat serum was similar and no variation was observed among the proteins on SDS-PAGE. Two secretory proteins of 55 and 65 kDa of both IVM oocytes and IVMFC embryos were observed on Western analysis. None of such proteins was observed in midluteal, estrus and early pregnant goat serum on western blotting. It can be concluded that IVM oocytes and IVMFC embryos secrete proteins in medium and two of them can develop antibody. The proteins secreted from embryos till morula stage was similar to that of oocytes. None of these oocyte/embryo released proteins were observed in blood circulation of estrus and early pregnant goats.

Effect of isoproturon on male reproductive system: clinical, histological and histoenzyonological studies in rats.

Isoproturon, a nonhalogenated substituted phenylurea herbicide, was evaluated for its cumulative toxic effects on testicular histomorphology., steroid hormone biosynthesis-related enzymes, spermatogenesis and sperm cells in adult albino rats. The compound, suspended in refined groundnut oil, was administered (po) to rats for 10 weeks @ 0,200, 400 and 800 mg/kg/day for 6 days/week. Isoproturon, at 800 mg/kg dose, decreased epididymal sperm count and percentage of motile sperms and increased the percentage of morphologically abnormal sperm cells. At the same dose, diameter of seminiferous tubules was reduced, number of tubules per microscopic field was increased and the percentage of tubules with evidence of spermatogenesis decreased. However, the percentage of damaged tubules was increased with 400 and 800 mg/kg doses. Histoenzymological observations revealed dose-related reduction in the activities of glucose-6-phosphate dehydrogenase and delta 5-3 beta-hydroxy steroid dehydrogenase. Activity of 17 beta-hydroxy steroid dehydrogenase was not affected appreciably. Overall findings suggest that isoproturon, at high dose, impairs androgen biosynthetic process, affects spermatogenesis and induces maturational anomalies of sperm cells in rat.

Subacute toxicity of urea herbicide, isoproturon, in male rats.

Isoproturon, a substituted phenylurea herbicide, was evaluated for its cumulative toxic effect on growth, organ weight and histomorphology of different organs in adult male rats. The compound (200, 400 and 800 mg/kg/day for 6 days/week), suspended in refined groundnut oil, was administered to rats p.o. for 7 and 10 weeks. There were no definite signs and symptoms of toxicity in treated rats but the herbicide significantly decreased the weekly body weight of rats at 800 mg/kg dose. Isoproturon, in all the three doses, increased the weight of liver in a dose-dependent manner. At 800 mg/kg dose, isoproturon increased the weight of kidney and heart, and decreased the weight of epididymis. Histopathological alterations in the organs were dose-dependent. Salient microscopic changes induced by isoproturon were hepatocellular degenerative changes and focal necrosis in liver, glomerular and tubular degeneration in kidney and haemosiderosis in spleen. Testis showed degeneration and desquamation of cells of germinal layers. Tubular lumens of testis and epididymis exhibited reduced number of spermatids and spermatozoa, respectively, indicating retardation of spermatogenesis.