Use of a highly specific monoclonal antibody against the central variable amino acid sequence of mammalian gonadotropin releasing hormone to evaluate GnRH-I tissue distribution compared with GnRH-I binding sites in adult male rats

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Abstract

PROBLEM: Recent evidence shows the existence of numerous isoforms of gonadotropin releasing hormone (GnRH), with high sequence homology and a core variable region. This raises the issue that previous GnRH distribution studies may have identified a variety of isoforms. This investigation was carried out to confirm the distribution and binding activity of GnRH-I only.

METHOD OF STUDY: A monoclonal antibody (7B101D10), with specificity for the core region of GnRH-I was used to stain formalin-fixed tissue sections from adult male Sprague-Dawley rats, while a biotinylated GnRH-I sequence was used with avidin-labelled HRP to evaluate regions of GnRH-I binding.

RESULTS AND CONCLUSIONS: GnRH-I expression was only found in the hypothalamus, cerebellum, anterior/fore brain and in Sertoli cells, while, binding activity was only present in the pituitary, subendocardium and subepicardium, thymic lymphocytes, peripheral blood lymphocytes and neutrophils. There was overlap in the olfactory neurons, liver (Kupffer macrophages and hepatocytes), spleen (lymphocytes and dendritic cells), myocardium and testes (spermatozoa and Leydig cells) and this may be further evidence of the paracrine/autocrine activity of a neuropeptide.

Original languageEnglish
Pages (from-to)239-48
Number of pages10
JournalAmerican Journal of Reproductive Immunology
Volume49
Issue number4
Early online date25 Mar 2003
DOIs
Publication statusPublished - 30 Apr 2003

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Tissue Distribution
Gonadotropin-Releasing Hormone
Rats
Amino Acid Sequence
Binding Sites
Monoclonal Antibodies
Tissue
Amino Acids
Lymphocytes
Protein Isoforms
Anterior Hypothalamus
Avidin
Leydig Cells
Sertoli Cells
Macrophages
Sequence Homology
Neuropeptides
Liver
Dendritic Cells
Cerebellum

Keywords

  • amino acid sequence
  • animals
  • antibodies, monoclonal
  • antibody specificity
  • binding sites
  • enzyme-linked immunosorbent assay
  • gonadotropin-releasing hormone
  • immunohistochemistry
  • male
  • protein isoforms
  • rats
  • receptors, LHRH
  • comparative study

Cite this

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title = "Use of a highly specific monoclonal antibody against the central variable amino acid sequence of mammalian gonadotropin releasing hormone to evaluate GnRH-I tissue distribution compared with GnRH-I binding sites in adult male rats",
abstract = "PROBLEM: Recent evidence shows the existence of numerous isoforms of gonadotropin releasing hormone (GnRH), with high sequence homology and a core variable region. This raises the issue that previous GnRH distribution studies may have identified a variety of isoforms. This investigation was carried out to confirm the distribution and binding activity of GnRH-I only.METHOD OF STUDY: A monoclonal antibody (7B101D10), with specificity for the core region of GnRH-I was used to stain formalin-fixed tissue sections from adult male Sprague-Dawley rats, while a biotinylated GnRH-I sequence was used with avidin-labelled HRP to evaluate regions of GnRH-I binding.RESULTS AND CONCLUSIONS: GnRH-I expression was only found in the hypothalamus, cerebellum, anterior/fore brain and in Sertoli cells, while, binding activity was only present in the pituitary, subendocardium and subepicardium, thymic lymphocytes, peripheral blood lymphocytes and neutrophils. There was overlap in the olfactory neurons, liver (Kupffer macrophages and hepatocytes), spleen (lymphocytes and dendritic cells), myocardium and testes (spermatozoa and Leydig cells) and this may be further evidence of the paracrine/autocrine activity of a neuropeptide.",
keywords = "amino acid sequence, animals, antibodies, monoclonal, antibody specificity, binding sites, enzyme-linked immunosorbent assay, gonadotropin-releasing hormone, immunohistochemistry, male, protein isoforms, rats, receptors, LHRH, comparative study",
author = "Khan, {Mohammad A H} and Ferro, {Valerie A} and Stimson, {William H}",
year = "2003",
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doi = "10.1034/j.1600-0897.2003.01202.x",
language = "English",
volume = "49",
pages = "239--48",
journal = "American Journal of Reproductive Immunology",
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T1 - Use of a highly specific monoclonal antibody against the central variable amino acid sequence of mammalian gonadotropin releasing hormone to evaluate GnRH-I tissue distribution compared with GnRH-I binding sites in adult male rats

AU - Khan, Mohammad A H

AU - Ferro, Valerie A

AU - Stimson, William H

PY - 2003/4/30

Y1 - 2003/4/30

N2 - PROBLEM: Recent evidence shows the existence of numerous isoforms of gonadotropin releasing hormone (GnRH), with high sequence homology and a core variable region. This raises the issue that previous GnRH distribution studies may have identified a variety of isoforms. This investigation was carried out to confirm the distribution and binding activity of GnRH-I only.METHOD OF STUDY: A monoclonal antibody (7B101D10), with specificity for the core region of GnRH-I was used to stain formalin-fixed tissue sections from adult male Sprague-Dawley rats, while a biotinylated GnRH-I sequence was used with avidin-labelled HRP to evaluate regions of GnRH-I binding.RESULTS AND CONCLUSIONS: GnRH-I expression was only found in the hypothalamus, cerebellum, anterior/fore brain and in Sertoli cells, while, binding activity was only present in the pituitary, subendocardium and subepicardium, thymic lymphocytes, peripheral blood lymphocytes and neutrophils. There was overlap in the olfactory neurons, liver (Kupffer macrophages and hepatocytes), spleen (lymphocytes and dendritic cells), myocardium and testes (spermatozoa and Leydig cells) and this may be further evidence of the paracrine/autocrine activity of a neuropeptide.

AB - PROBLEM: Recent evidence shows the existence of numerous isoforms of gonadotropin releasing hormone (GnRH), with high sequence homology and a core variable region. This raises the issue that previous GnRH distribution studies may have identified a variety of isoforms. This investigation was carried out to confirm the distribution and binding activity of GnRH-I only.METHOD OF STUDY: A monoclonal antibody (7B101D10), with specificity for the core region of GnRH-I was used to stain formalin-fixed tissue sections from adult male Sprague-Dawley rats, while a biotinylated GnRH-I sequence was used with avidin-labelled HRP to evaluate regions of GnRH-I binding.RESULTS AND CONCLUSIONS: GnRH-I expression was only found in the hypothalamus, cerebellum, anterior/fore brain and in Sertoli cells, while, binding activity was only present in the pituitary, subendocardium and subepicardium, thymic lymphocytes, peripheral blood lymphocytes and neutrophils. There was overlap in the olfactory neurons, liver (Kupffer macrophages and hepatocytes), spleen (lymphocytes and dendritic cells), myocardium and testes (spermatozoa and Leydig cells) and this may be further evidence of the paracrine/autocrine activity of a neuropeptide.

KW - amino acid sequence

KW - animals

KW - antibodies, monoclonal

KW - antibody specificity

KW - binding sites

KW - enzyme-linked immunosorbent assay

KW - gonadotropin-releasing hormone

KW - immunohistochemistry

KW - male

KW - protein isoforms

KW - rats

KW - receptors, LHRH

KW - comparative study

U2 - 10.1034/j.1600-0897.2003.01202.x

DO - 10.1034/j.1600-0897.2003.01202.x

M3 - Article

VL - 49

SP - 239

EP - 248

JO - American Journal of Reproductive Immunology

JF - American Journal of Reproductive Immunology

SN - 1046-7408

IS - 4

ER -