Functional analysis of α1 adrenergic receptor using mutant mice generated by the gene targeting.

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α1-ARs play critical roles in the regulation of a variety of physiological processes. Increasing evidence suggests that these physiological responses are regulated by multiple receptor subtypes that are structurally homologous. Molecular cloning identified three distinct cDNA encoding α1-AR subtypes (α1a, α1b, α1d). Among the α1-AR subtypes, the function of the α1D-AR remains uncertain, although it is expressed in various tissues. In order to examine the physiological role and the structure-function relationship, we cloned and characterized a gene for the murine α1D-AR. Using a rat α1D-AR cDNA as a probe, the gene for the murine α1D-AR gene is cloned from murine genomic libraries. It consists of two exons and an intron which is over 5 kilobases and interrupts the coding region of the sixth trasmembrane domain. With these information and DNA fragments, a targeting vector for disrupting the gene in mice was constructed and transfected into ES cells. Several positive clones were isolated which were used for blastocyteinjenction. Homozygous mutant mice for α1D-AR were generated. They look normal and proved to be fertile. These mice should be useful for analysis for interaction among α1-ARs.