Identification and cloning of site C splice variants of plasma membrane Ca-ATPase in the gerbil cochlea.
Plasma membrane Ca-ATPase (PMCA) gene products were identified in the gerbil cochlea by reverse-transcription polymerase chain reaction (RT-PCR). Cochlear cDNA was amplified using PMCA isoform-specific primers from splice site C, the calmodulin binding domain. PCR products were cloned and sequenced. The putative housekeeping PMCA genes, 1b and 4b, as expected, were present in the gerbil cochlea and shared 98.6 and 100% amino acid homology with published rat sequences, at splice site C, respectively. PMCA2b, 3a and 3b splice variants also were detected in cochlear cDNAs and shared 95, 94.3 and 98% amino acid homology with their rat counterparts. PMCA isoforms 2 and 3 have been shown to occur in highly specialized tissues, such as muscle and brain, that require finely tuned regulation of intracellular free Ca2+ levels. The presence of several isoforms and splice variants of PMCA in the cochlea most probably reflects their differential expression among the several cell types that have been shown to contain immunoreactive PMCA. This suggests that the cochlea has developed complex mechanisms that finely tune intracellular Ca2+ levels in different highly specialized cell types.
Crouch
J J
JJ
Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29425, USA. crouchjj@musc.edu
Schulte
B A
BA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D003001:Cloning, Molecular; D003051:Cochlea / Q000201:enzymology*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005849:Gerbillinae; D006706:Homeostasis; D017136:Ion Transport; D007527:Isoenzymes; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D019154:Protein Splicing; D051381:Rats; D017386:Sequence Homology, Amino Acid
D002147:Calmodulin; D018076:DNA, Complementary; D007527:Isoenzymes; D000252:Calcium-Transporting ATPases; D002118:Calcium