Characterization of the luteinizing hormone response to continuous infusions of gonadotropin releasing hormone using perifused pituitaries from intact, ovariectomized and steroid-treated rats.

D. M. Baldwin, Jacques W Ramey, W. W. Wilfinger

Research output: Contribution to journalArticle

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Abstract

Anterior pituitary glands obtained from rats at various stages of the estrous cycle and from short-term ovariectomized (OVX) rats with or without physiological replacement of estradiol-17 beta (E2) and/or progesterone (P), were perifused in vitro with continuous infusions (4 h) of gonadotropin releasing hormone (GnRH) (12 ng/h). In all treatment groups the in vitro pattern of luteinizing hormone (LH) release in response to GnRH was characterized by an initial low rate of LH release (initial phase; 20-70 min) followed by a significant augmented rate of secretion during the late phase (120-240 min) with the exception of the estrous and OVX + P groups in which LH was released at a constant rate. The total amount of LH released in response to GnRH was similar for glands removed on estrus, diestrus-I (D-I) and D-II, but increased significantly for glands removed on 0900 h proestrus with a further increase at 1400 h proestrus. Ovariectomy reduced the total LH released in vitro by 50-60% and 85-90% compared with estrous, D-I, D-II and proestrous groups, respectively. In vivo treatment of OVX rats with E2 restored the in vitro LH response to levels comparable to those in the 0900 h proestrous group while treatment with P + E2 further increased the rate of LH release in the initial phase to levels similar to those observed in the 1400 h proestrous group. In all treatment groups, addition of 5 microM cycloheximide to the perifusion media significantly inhibited GnRH-stimulated LH release during the late phase without significantly altering the initial LH response except in the OVX + E2P group in which it was partially inhibited. These results demonstrate that perifused pituitaries maintain their characteristic responsiveness to GnRH in vitro. Furthermore, they indicate that LH secretion in response to continual GnRH stimulation involves protein synthesis-independent and -dependent components. E2, in vivo, enhances the magnitude of both components in response to GnRH in vitro, while E2 + P may further enhance the initial in vitro response to GnRH through a protein synthesis-dependent mechanism.

Original languageEnglish (US)
Pages (from-to)99-111
Number of pages13
JournalBiology of reproduction
Volume29
Issue number1
DOIs
StatePublished - Jan 1 1983

Fingerprint

Luteinizing Hormone
Gonadotropin-Releasing Hormone
Steroids
Diestrus
Proestrus
Anterior Pituitary Gland
Estrous Cycle
Estrus
Ovariectomy
Cycloheximide
In Vitro Techniques
Progesterone
Estradiol
Proteins

ASJC Scopus subject areas

  • Cell Biology

Cite this

Characterization of the luteinizing hormone response to continuous infusions of gonadotropin releasing hormone using perifused pituitaries from intact, ovariectomized and steroid-treated rats. / Baldwin, D. M.; Ramey, Jacques W; Wilfinger, W. W.

In: Biology of reproduction, Vol. 29, No. 1, 01.01.1983, p. 99-111.

Research output: Contribution to journalArticle

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abstract = "Anterior pituitary glands obtained from rats at various stages of the estrous cycle and from short-term ovariectomized (OVX) rats with or without physiological replacement of estradiol-17 beta (E2) and/or progesterone (P), were perifused in vitro with continuous infusions (4 h) of gonadotropin releasing hormone (GnRH) (12 ng/h). In all treatment groups the in vitro pattern of luteinizing hormone (LH) release in response to GnRH was characterized by an initial low rate of LH release (initial phase; 20-70 min) followed by a significant augmented rate of secretion during the late phase (120-240 min) with the exception of the estrous and OVX + P groups in which LH was released at a constant rate. The total amount of LH released in response to GnRH was similar for glands removed on estrus, diestrus-I (D-I) and D-II, but increased significantly for glands removed on 0900 h proestrus with a further increase at 1400 h proestrus. Ovariectomy reduced the total LH released in vitro by 50-60{\%} and 85-90{\%} compared with estrous, D-I, D-II and proestrous groups, respectively. In vivo treatment of OVX rats with E2 restored the in vitro LH response to levels comparable to those in the 0900 h proestrous group while treatment with P + E2 further increased the rate of LH release in the initial phase to levels similar to those observed in the 1400 h proestrous group. In all treatment groups, addition of 5 microM cycloheximide to the perifusion media significantly inhibited GnRH-stimulated LH release during the late phase without significantly altering the initial LH response except in the OVX + E2P group in which it was partially inhibited. These results demonstrate that perifused pituitaries maintain their characteristic responsiveness to GnRH in vitro. Furthermore, they indicate that LH secretion in response to continual GnRH stimulation involves protein synthesis-independent and -dependent components. E2, in vivo, enhances the magnitude of both components in response to GnRH in vitro, while E2 + P may further enhance the initial in vitro response to GnRH through a protein synthesis-dependent mechanism.",
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