Gene targeting of natriuretic peptide family and their functional analyzes

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Natriuretic peptide family is comprised of three peptides (ANP,BNP,CNP). These peptides bind receptor quanylate cyclase (GC-A and GC-B), increase cGMP concentration, and alter cell functions. ANP and BNP mainly function as cardiac hormones regulating blood pressure, while CNP is a neuropeptide in brain or a local regulator in the vascular wall. In order to elucidate their in vivo functions and estimate the contribution to blood pressure regulation, we have planned to establish a mouse which is deleted the natriuretic peptide genes by using the newly developed gene targeting method.We cloned mouse ANP,BNP and CNP gene of 5-6 kb containing complete sets of each exons. About 3-6 kb sequence of each gene was determine. In the case of CNP gene, the third exon region was first sequenced by the present study. In addition to three natriuretic peptide genes, we further cloned mouse GC-A and GC-B gene to analyze their functions. Portions of GC-A and GC-B gene structure containing functional domains are now being determined. By a series of these studies, appropriate fragments used for the gene targeting of the natriuretic peptides and their receptors have been obtained, and most of the sequences have already determined.