A cloned chromogranin A (CgA) cDNA detects a 2.3Kb mRNA in diverse neuroendocrine tissues.
CgA is a 72Kd protein of unknown function that is present in many neuroendocrine tissues and co-secreted with their resident hormones. We prepared a cDNA library to the mRNA from CgA-producing human medullary thyroid carcinoma (MTC) cells in the expression vector lambda gt11. The library was screened with a panel of one polyclonal and two monoclonal antibodies to CgA. The specificity of the antibodies for CgA was demonstrated by immunoassay, immunohistology, and immunoprecipitation of the in vitro translation products of mRNA from CgA-producing tissues. A chromogenic second antibody identified five immunoreactive clones. Their cDNA inserts were isolated after EcoRI digestion and agarose gel electrophoresis. These cDNAs were 32P-labelled and used as probes in Northern hybridization studies. An mRNA of approximately 2.3Kb was detected with the cDNA probes in human cell lines from MTC and lung cancers that were shown to produce CgA and in human pheochromocytoma and bovine adrenal medulla tissue. To confirm its identity, one of the putative CgA cDNAs was subcloned into a plasmid and was used to hybridization-arrest the in vitro translation of CgA mRNA. These studies demonstrate the cloning of cDNAs which hybridize with CgA mRNA from diverse neuroendocrine tissues.
D000313:Adrenal Medulla / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D002277:Carcinoma / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D053379:Chromogranin A; D002864:Chromogranins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics*; D010673:Pheochromocytoma / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D011863:Radioimmunoassay; D013964:Thyroid Neoplasms / Q000378:metabolism
D000911:Antibodies, Monoclonal; C506150:CHGA protein, human; D053379:Chromogranin A; D002864:Chromogranins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger