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Endocrinology, Vol 119, 1510-1518, Copyright © 1986 by Endocrine Society

ARTICLES

Desensitization to gonadotropin-releasing hormone in perifused chicken anterior pituitary cells

JA King, JS Davidson and RP Millar

A perifusion method consisting of dispersed chicken anterior pituitary cells suspended in columns of Bio-Gel was developed to monitor the dynamics of LH release. The perifused cells responded to chicken I GnRH (Gln8-GnRH) in a dose-dependent manner. The ED50 was 3 X 10(-10) M, and maximal LH release occurred in response to 4 X 10(-9) M Gln8-GnRH. Continuous administration of 10(-7) M Gln8-GnRH and agonist stimulated an initial 8- to 10-fold increase in LH release within minutes. LH release then declined rapidly, reaching basal levels within 100 min. A biphasic response was noted. Calcium ionophore A23187 was effective in releasing additional LH from cells desensitized to 10(-7) Gln8-GnRH and agonist, indicating that total cellular LH was not depleted. In contrast, delivery of 2-min pulses of 10(-7) M and 10(-9) M Gln8-GnRH at a frequency of one pulse every 30 or 60 min for 3-5 h maintained pituitary responsiveness. Exposure to 10(-7) M Gln8-GnRH for 20 min was sufficient to desensitize pituitary cells to subsequent Gln8-GnRH stimulation. However, 20-min exposure to 10(-7) M GnRH antagonist neither evoked LH release nor had a desensitizing effect on subsequent stimulation by 10(-7) M Gln8-GnRH, indicating that receptor activation, not merely receptor binding, is necessary for Gln8-GnRH-mediated homologous desensitization. Pituitary cells desensitized by 20-min exposure to 10(-8) M Gln8-GnRH maintained responsiveness to a higher dose (10(-6 M) of Gln8-GnRH, suggesting that down-regulation of pituitary GnRH receptors might play a part in desensitization. Calcium ionophore A23187 partially desensitized pituitary cells to subsequent stimulation with Gln8-GnRH, probably due to depletion of releasable LH or desensitization of calcium-coupled secretory mechanisms. In calcium- free medium, 10(-7) M Gln8-GnRH did not evoke LH release, but nevertheless partially desensitized cells to subsequent 10(-7) M Gln8- GnRH stimulation. Thus desensitization is partially calcium-dependent. These findings demonstrate that the GnRH-mediated desensitization of gonadotrophs is a characteristic of chicken pituitary cells as in the mammal. However, chicken pituitary cells differ from mammalian cells in that desensitization is more rapid and partially dependent on extracellular calcium.


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C. K. Cheng and P. C. K. Leung
Molecular Biology of Gonadotropin-Releasing Hormone (GnRH)-I, GnRH-II, and Their Receptors in Humans
Endocr. Rev., April 1, 2005; 26(2): 283 - 306.
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