IN-VITRO CELL-CULTURE AND ADRENOCORTICOTROPIN SECRETION BY INDIAN BLACKBUCK ANTELOPE (ANTILOPE CERVICAPRA) ANTERIOR-PITUITARY GLANDS COLLECTED UNDER FIELD CONDITIONS

The objective of this study was to collect pituitary glands from India n blackbuck antelope (Antilope cervicapra) in the field, transport the tissue to a laboratory, and establish a primary cell culture to deter mine the viability and responsiveness of the cells to two hypothalamic neuropeptides, corticotropin-releasing factor (CRF) and vasopressin ( VP). Blood samples were collected and pituitary glands (n = 3) were we ighed and placed in a transport medium cooled to 4 degrees C. Anterior pituitaries were enzymatically dissociated by collagenase treatment. As determined by trypan blue exclusion, 76.6% of the isolated cells we re viable. Cells (2.0 x 10(5) viable cells/well) were plated and rando mly assigned (six wells/treatment) to one of four treatments on day 6: control (medium alone), 0.1 mu M bovine CRF, 1 mu M Vf, or a combinat ion of 0.1 mu M CRF and 1.0 mu M VP, or to one of two treatments on da y 7: control or 0.1 mu M bovine CRF (three wells/treatment). Following a 4-hr incubation period, medium was collected and stored at -20 degr ees C until concentration of adrenocorticotropin (ACTH) was determined by radioimmunoassay. On day 6 ofculture, medium concentration of ACTH increased 5.6- and 6.3-fold (P < 0.000 1) following a 4-hr incubation with 0.1 mu M CRF and 1 mu M VP, respectively, relative to control (2 3.5 +/- 1.2 pg/ml). CRF- and VP-treated wells did not differ (P > 0.10 ) in medium concentration of ACTH on day 6. Treatment with CRF + VP in creased (P < 0.03) medium concentration of ACTH 10.2-, 1.7-, and 1.5-f old relative to control, CRF, or VP groups, respectively. Medium conce ntration of ACTH from control wells did not differ (P > 0.10) between days 6 and 7 of culture. CRF treatment increased (P < 0.01) medium con centration of ACTH by 6.8-fold relative to the control on day 7. This study outlines a method for the successful in vitro culture of anterio r pituitary cells from Indian blackbuck antelope and demonstrates the viability and responsiveness of the cells following held harvest, coll ection, cooling, and transportation of tissue.