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        In Vitro Studies on the Release of Intracelluar Prolactin from Lymphocytes Using Strees Related Amines and Hormones

        Sharma, G.T.,Majumdar, A.C.,Gupta, L.K. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.7

        Circulating lymphocytes collected from control and heat-stressed buffaloes were subjected to in vitro culture with glucocorticoids, epinephrine or serotonin and their effect, if any, on the release of intracellular prolactin (PRL) was studied using ELISA and C-ELISA techniques. It was noted from the study that PRL level was higher in lymphocytes than in plasma of the control and heat-stressed animals, and that the PRL levels increased in the plasma of heat-stressed animals compared to that of non stressed animals with a significant decrease in lymphocytic PRL content by heat stress. Epinephrine and serotonin significantly increased the release of intracellular PRL from the lymphocytes of both in the control and the heat-stressed buffaloes but release of PRL from lymphocyte was not significantly changed by cortisol treatment in both control and heat-stressed buffaloes as compared to epinephrine and serotonin in vitro. When lympocytes were incubated with serotonin, it caused drastic lysis of the lymphocytes but epinephirine and cortisol did not show any lysis. It may be concluded from this study that hormones like epinephrine or serotonin known to increase during stress, release intracellular PRL from lymphocytes, the satellite PRL storage/synthesizing organ of blood, although the mechanism of the release is different.

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        Secretory Proteins from Goat Oocytes Matured in Culture

        Malakar, Dhruba,Majumdar, A.C. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.3

        In this experiment, oocytes were collected from goat ovaries available in slaughterhouse by follicle puncture method. Morphologically culturable type of oocytes which having compact, multilayered cumulus granulosa cell complex and evenly granulated cytoplasm, was separated under a stereozoom microscope. Oocytes were washed thoroughly in maturation medium containing TCM-199, $1{\mu}g/ml$ estradiol-$17{\beta}$, 0.5 ${\mu}g/ml$ FSH, $100{\mu}g/ml$ LH, 3 mg/ml BSA and 10% estrus goat serum. Washed oocytes were cultured into maturation medium on granulosa cell monolayer. Culture plate was then kept into $CO_2$ incubator at $38{\pm}1^{\circ}C$, maximum humidity and 5% $CO_2$ for 18 h. After maturation the oocytes were washed thoroughly with maturation medium containing polyvinyl alcohol (PVA) without serum and BSA and further cultured for 12 h for secretory proteins of oocytes. PVA medium was collected, pooled and concentrated by 5000 cut off centrisart. Secretory proteins were separated on 12.5% SDS-PAGE. A total number of 3.41 oocytes per ovary were obtained and 2.17 culturable oocytes per ovary were cultured into maturation medium. After 18 h of maturation, 4,567 oocytes (1.82 oocytes per ovary) were further cultured into serum and BSA free PVA medium for its secretory proteins. Four secretory proteins of oocytes with approximately molecular weight of 45, 55, 65 and 95 kDa were obtained on SDS-PAGE in silver staining and three proteins with approximately molecular weight of 45, 55 and 65 kDa in Coomassie brilliant blue staining. In conclusion, four secretory proteins with approximately molecular weight of 45, 55, 65 and 95 kDa was obtained from in vitro cultured oocytes of goats.

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        Meiotic Competence of Caprine Oocytes During IVM on Granulosa Cell Monolayers Developed from Small and Large Follicles in Comparison to the Granulosa Cell Coculture

        Sharma, G. Taru,Teotia, Alok,Majumdar, A.C. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.6

        Evaluation of the granulosa cell (GC) monolayers developed from small (<5 mm) and large (>5 mm) follicles on the meiotic competence of caprine oocytes during in vitro maturation was done in this study in comparison to the granulosa cell coculture. Ovaries were collected from the local abattoir and follicular contents were aspirated for the monolayer culture. For IVM the oocytes were collected by puncturing the nonatretic follicles (>4 mm). Results revealed that at the same seeding rate, small follicular granulosa cell monolayer achieved confluence 24-48 h earlier than large follicular granulosa cell monolayer. GC monolayers significantly p (<0.05) improved the rate of meiotic resumption and nuclear maturation (84.76% vs 74.74%) after 27 h of culture in comparison to GC coculture. Statistically there was no significant difference in the maturation rate between the caprine oocytes matured over small or large follicular GC monolayers. It is concluded from the present study that GC monolayers support better nuclear and cytoplasmic maturation of growing caprine oocytes which is evident by better maturation rate over GC monolayer as compared to the oocytes matured with GC coculture. Granulosa cells from small and large follicles can be used for IVM with more or less in the same efficiency after conditioning them with maturation media in 18-24 h before the onset of culture.

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