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
Article first published online: 25 MAR 2003
American Journal of Reproductive Immunology
Volume 49, Issue 4, pages 239–248, April 2003
How to Cite
Khan, M. A. H., Ferro, V. A. and Stimson, W. H. (2003), 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. American Journal of Reproductive Immunology, 49: 239–248. doi: 10.1034/j.1600-0897.2003.01202.x
- Issue published online: 25 MAR 2003
- Article first published online: 25 MAR 2003
- Submitted July 31, 2002; revised September 10, 2002; accepted September 17, 2002.
- GnRH-I tissue distribution;
- monoclonal antibodies;
- variable and core region sequences
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.