Perifusion model system to culture bovine hypothalamic slices in series with dispersed anterior pituitary cells |
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Authors: | H A Hassan R A Merkel |
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Institution: | (1) Present address: Lilly Research Laboratories, P.O. Box 708, 46140 Greenfield, Indiana;(2) Growth Biology Program, Department of Animal Science, and Food Science and Human Nutrition, Michigan State University, 48823 East Lansing, Michigan |
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Abstract: | Summary Dispersed bovine anterior pituitary cells were incubated either in static or perifusion cultures to assess basal growth hormone
release as well as stimulatory and inhibitory effects of growth hormone-releasing hormone and somatostatin, respectively,
on growth hormone release. Total concentrations of growth hormones over a 12-hour incubation period were fivefold greater
in perifused than in static cultures (2034 ± 160 vs. 387 ± 33 ng/12 h). A dose-dependent increase in growth hormone secretion
in response to challenge with growth hormone-releasing hormone (10−12 to 10−8
M) for 1 h was observed in both static and perifusion cultures; however, perifused cells were more responsive to the same concentration
of neuropeptide than those in static culture. Concentrations of somatostatin (10−12 to 10−8
M) for 1 h did not inhibit basal growth hormone secretion in either static or perifusion cultures. To establish model, slices
of the hypothalamus, immediately adjacent to the sagittal midline, were perifused in series with anterior pituitary cells,
and media effluent was assayed for growth hormone concentrations. Release of growth hormone was pulsatile and seemed to mimic
the episodic pattern of bovine secretion. Hypothalamic slices were placed in one chamber of the perifusion system, and basal
secretion of growth hormone-releasing hormone and somatostatin was pulsatile in media effluent. Tissue viability of hypothalamic
slices and anterior pituitary cells was evaluated by KCl depolarization. Tissues were viable for at least 120 h. Thus, this
hypothalamo-pituitary dual chamber perifusion system is a valid in vitro model to study regulation of growth hormone secretion. |
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Keywords: | perifusion system hypothalamic slices anterior pituitary cells bovine |
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