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 language | English |
|---|---|
| Pages (from-to) | 239-48 |
| Number of pages | 10 |
| Journal | American Journal of Reproductive Immunology |
| Volume | 49 |
| Issue number | 4 |
| Early online date | 25 Mar 2003 |
| DOIs | |
| Publication status | Published - 30 Apr 2003 |
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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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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. / Khan, Mohammad A H; Ferro, Valerie A; Stimson, William H.
In: American Journal of Reproductive Immunology, Vol. 49, No. 4, 30.04.2003, p. 239-48.Research output: Contribution to journal › Article
TY - JOUR
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 -