Study summary · research use only
A monoclonal antibody raised against a synthetic oxytocin peptide stains mouse hypothalamic neurones
Plain-language summary
Paraphrased from the published abstract below — not a verdict on whether anything works.
In this mouse study, a monoclonal antibody (7a5) against oxytocin was generated from hybridoma cells derived from myeloma cells and spleen lymphocytes of mice immunised with a synthetic oxytocin peptide. The 7a5 antibody bound oxytocin in ELISA, and diluted growth medium (up to 5000-fold) was used for immunohistochemistry. The abstract reports no 7a5 immunoreactivity in the paraventricular nucleus of oxytocin knockout mice, while this region stained strongly with an anti-vasopressin antibody. In wild-type mice, 7a5 staining in the paraventricular and supraoptic nuclei was reported as similar to staining with anti-oxytocin antibodies, with most 7a5-stained cells not co-stained by the vasopressin antibody, and the suprachiasmatic nucleus stained by the vasopressin antibody but not 7a5. The authors describe 7a5 as a new anti-oxytocin monoclonal antibody recognising oxytocin and not vasopressin.
Abstract
A monoclonal antibody against oxytocin was generated in 7a5 hybridoma cells derived from myeloma cells and lymphocytes from the spleen of mice immunised with a synthetic oxytocin peptide. The 7a5 monoclonal antibody bound with oxytocin in enzyme-linked immunosorbent assays. 7a5 cell growth medium was diluted up to 5000-fold and used for immunohistochemistry. First, to test the specificity of the 7a5 antibody against oxytocin, we stained brain tissues of oxytocin knockout mice, comprising mice in which the first exon of the oxytocin-neurophysin gene is deleted. No 7a5 immunoreactivity was detected in the paraventricular nucleus (PVN) of the hypothalamus of oxytocin knockout mice; however, this area was strongly stained with the anti-vasopressin polyclonal antibody, HM07. Tissue preparations of the wild-type mouse PVN and supraoptic nucleus (SON) displayed 7a5 immunoreactivity that was indistinguishable from the staining produced with an anti-oxytocin polyclonal antibody, HM06. The immunoreactivity of HM06 in the PVN was similar to that of an anti-oxytocin monoclonal antibody, PS38. We then examined the cross-reactivity of 7a5 with arginine vasopressin. The majority of cell soma and processes stained by 7a5 were not co-stained with the vasopressin antibody in SON and PVN regions. Furthermore, the suprachiasmatic nucleus was stained by the vasopressin antibody but not by 7a5. These results demonstrate that 7a5 is a new anti-oxytocin monoclonal antibody recognising oxytocin and not vasopressin; therefore, 7a5 can be used to investigate the role of oxytocin in the brain.
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