Effects of ovariectomy on GnRH mRNA, prognrh and GnRH levels in the preoptic hypothalamus of the female rat

Martin Kelly, James Garrett, Martha A. Bosch, Charles Roselli, James Douglass, John Adelman, Oline Ronnekleiv

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Experiments were carried out to investigate the effects of ovariectomy on gonadotropin-releasing hormone (GnRH) messenger RNA (mRNA), proGnRH and GnRH peptide levels in the hypothalamus of remale rats. Intact proestrous female rats and female rats, which had been ovariectomized for 2 weeks, were sacrificed at 9.00 h and the preoptic area (POA) and basal hypothalamus (BH) were dissected out and frozen on dry ice. One group of tissues from proestrous control and ovariectomized females were extracted in acetic acid, centrifuged at 13,000 gand the supernatant purified on a C18 column. The purified extract was then radioimmunoassayed for proGnRH, using a specific antiserum to rat proGnRH (ARK-2), and for GnRH using the El-14 antiserum. Total cellular RNA was isolated from another group of tissues and prepared as Northern blots. Hybridization with 32P-labeled GnRH cRNA was used to detect GnRH mRNA. A third group of proestrous and ovariectomized female rats were perfused, and 50 μm vibratome sections were cut. These were immunos-tained with proGnRH or GnRH antiserum, followed by in situ hybridization with J5S-labeled GnRH cRNA to detect GnRH mRNA. Based on the histochemical staining, mRNA was colocalized to the cell soma of neurons containing proGnRH and GnRH throughout the POA and BH. Based on the radioimmunoassay, proGnRH levels were 2 times higher in the POA versus the BH, but GnRH levels were 6-7 times higher in the BH. Ovariectomy significantly decreased proGnRH levels in both the POA and BH, while GnRH decreased in the BH. In contrast, quantitative Northern blot analysis demonstrated that ovariectomy had no effect on mRNA levels in the POA and BH. These data indicate that the effects of ovariectomy on proGnRH and GnRH levels are a result of altered translation, posttranslational processing and/or secretion of GnRH.

Original languageEnglish (US)
Pages (from-to)88-97
Number of pages10
JournalNeuroendocrinology
Volume49
Issue number1
DOIs
StatePublished - 1989

Fingerprint

Ovariectomy
Gonadotropin-Releasing Hormone
Hypothalamus
Messenger RNA
Preoptic Area
Immune Sera
Complementary RNA
Northern Blotting
progonadoliberin I
Dry Ice
Carisoprodol
Acetic Acid
Radioimmunoassay
In Situ Hybridization
RNA

Keywords

  • Basal hypothalamus
  • GnRH
  • mRNA
  • Ovariectomy
  • Preoptic area
  • Proestrous
  • proGnRH

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism
  • Cellular and Molecular Neuroscience
  • Endocrine and Autonomic Systems
  • Neuroscience(all)

Cite this

Effects of ovariectomy on GnRH mRNA, prognrh and GnRH levels in the preoptic hypothalamus of the female rat. / Kelly, Martin; Garrett, James; Bosch, Martha A.; Roselli, Charles; Douglass, James; Adelman, John; Ronnekleiv, Oline.

In: Neuroendocrinology, Vol. 49, No. 1, 1989, p. 88-97.

Research output: Contribution to journalArticle

@article{c533dbdfe8b746e6be5d6500efee19af,
title = "Effects of ovariectomy on GnRH mRNA, prognrh and GnRH levels in the preoptic hypothalamus of the female rat",
abstract = "Experiments were carried out to investigate the effects of ovariectomy on gonadotropin-releasing hormone (GnRH) messenger RNA (mRNA), proGnRH and GnRH peptide levels in the hypothalamus of remale rats. Intact proestrous female rats and female rats, which had been ovariectomized for 2 weeks, were sacrificed at 9.00 h and the preoptic area (POA) and basal hypothalamus (BH) were dissected out and frozen on dry ice. One group of tissues from proestrous control and ovariectomized females were extracted in acetic acid, centrifuged at 13,000 gand the supernatant purified on a C18 column. The purified extract was then radioimmunoassayed for proGnRH, using a specific antiserum to rat proGnRH (ARK-2), and for GnRH using the El-14 antiserum. Total cellular RNA was isolated from another group of tissues and prepared as Northern blots. Hybridization with 32P-labeled GnRH cRNA was used to detect GnRH mRNA. A third group of proestrous and ovariectomized female rats were perfused, and 50 μm vibratome sections were cut. These were immunos-tained with proGnRH or GnRH antiserum, followed by in situ hybridization with J5S-labeled GnRH cRNA to detect GnRH mRNA. Based on the histochemical staining, mRNA was colocalized to the cell soma of neurons containing proGnRH and GnRH throughout the POA and BH. Based on the radioimmunoassay, proGnRH levels were 2 times higher in the POA versus the BH, but GnRH levels were 6-7 times higher in the BH. Ovariectomy significantly decreased proGnRH levels in both the POA and BH, while GnRH decreased in the BH. In contrast, quantitative Northern blot analysis demonstrated that ovariectomy had no effect on mRNA levels in the POA and BH. These data indicate that the effects of ovariectomy on proGnRH and GnRH levels are a result of altered translation, posttranslational processing and/or secretion of GnRH.",
keywords = "Basal hypothalamus, GnRH, mRNA, Ovariectomy, Preoptic area, Proestrous, proGnRH",
author = "Martin Kelly and James Garrett and Bosch, {Martha A.} and Charles Roselli and James Douglass and John Adelman and Oline Ronnekleiv",
year = "1989",
doi = "10.1159/000125095",
language = "English (US)",
volume = "49",
pages = "88--97",
journal = "Neuroendocrinology",
issn = "0028-3835",
publisher = "S. Karger AG",
number = "1",

}

TY - JOUR

T1 - Effects of ovariectomy on GnRH mRNA, prognrh and GnRH levels in the preoptic hypothalamus of the female rat

AU - Kelly, Martin

AU - Garrett, James

AU - Bosch, Martha A.

AU - Roselli, Charles

AU - Douglass, James

AU - Adelman, John

AU - Ronnekleiv, Oline

PY - 1989

Y1 - 1989

N2 - Experiments were carried out to investigate the effects of ovariectomy on gonadotropin-releasing hormone (GnRH) messenger RNA (mRNA), proGnRH and GnRH peptide levels in the hypothalamus of remale rats. Intact proestrous female rats and female rats, which had been ovariectomized for 2 weeks, were sacrificed at 9.00 h and the preoptic area (POA) and basal hypothalamus (BH) were dissected out and frozen on dry ice. One group of tissues from proestrous control and ovariectomized females were extracted in acetic acid, centrifuged at 13,000 gand the supernatant purified on a C18 column. The purified extract was then radioimmunoassayed for proGnRH, using a specific antiserum to rat proGnRH (ARK-2), and for GnRH using the El-14 antiserum. Total cellular RNA was isolated from another group of tissues and prepared as Northern blots. Hybridization with 32P-labeled GnRH cRNA was used to detect GnRH mRNA. A third group of proestrous and ovariectomized female rats were perfused, and 50 μm vibratome sections were cut. These were immunos-tained with proGnRH or GnRH antiserum, followed by in situ hybridization with J5S-labeled GnRH cRNA to detect GnRH mRNA. Based on the histochemical staining, mRNA was colocalized to the cell soma of neurons containing proGnRH and GnRH throughout the POA and BH. Based on the radioimmunoassay, proGnRH levels were 2 times higher in the POA versus the BH, but GnRH levels were 6-7 times higher in the BH. Ovariectomy significantly decreased proGnRH levels in both the POA and BH, while GnRH decreased in the BH. In contrast, quantitative Northern blot analysis demonstrated that ovariectomy had no effect on mRNA levels in the POA and BH. These data indicate that the effects of ovariectomy on proGnRH and GnRH levels are a result of altered translation, posttranslational processing and/or secretion of GnRH.

AB - Experiments were carried out to investigate the effects of ovariectomy on gonadotropin-releasing hormone (GnRH) messenger RNA (mRNA), proGnRH and GnRH peptide levels in the hypothalamus of remale rats. Intact proestrous female rats and female rats, which had been ovariectomized for 2 weeks, were sacrificed at 9.00 h and the preoptic area (POA) and basal hypothalamus (BH) were dissected out and frozen on dry ice. One group of tissues from proestrous control and ovariectomized females were extracted in acetic acid, centrifuged at 13,000 gand the supernatant purified on a C18 column. The purified extract was then radioimmunoassayed for proGnRH, using a specific antiserum to rat proGnRH (ARK-2), and for GnRH using the El-14 antiserum. Total cellular RNA was isolated from another group of tissues and prepared as Northern blots. Hybridization with 32P-labeled GnRH cRNA was used to detect GnRH mRNA. A third group of proestrous and ovariectomized female rats were perfused, and 50 μm vibratome sections were cut. These were immunos-tained with proGnRH or GnRH antiserum, followed by in situ hybridization with J5S-labeled GnRH cRNA to detect GnRH mRNA. Based on the histochemical staining, mRNA was colocalized to the cell soma of neurons containing proGnRH and GnRH throughout the POA and BH. Based on the radioimmunoassay, proGnRH levels were 2 times higher in the POA versus the BH, but GnRH levels were 6-7 times higher in the BH. Ovariectomy significantly decreased proGnRH levels in both the POA and BH, while GnRH decreased in the BH. In contrast, quantitative Northern blot analysis demonstrated that ovariectomy had no effect on mRNA levels in the POA and BH. These data indicate that the effects of ovariectomy on proGnRH and GnRH levels are a result of altered translation, posttranslational processing and/or secretion of GnRH.

KW - Basal hypothalamus

KW - GnRH

KW - mRNA

KW - Ovariectomy

KW - Preoptic area

KW - Proestrous

KW - proGnRH

UR - http://www.scopus.com/inward/record.url?scp=0024494243&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024494243&partnerID=8YFLogxK

U2 - 10.1159/000125095

DO - 10.1159/000125095

M3 - Article

VL - 49

SP - 88

EP - 97

JO - Neuroendocrinology

JF - Neuroendocrinology

SN - 0028-3835

IS - 1

ER -