Human Plasma Membrane Ca Research Articles

Studies on a PMCA-like protein in the outer mantle epithelium of Anodonta cygnea: insights on calcium transcellular dynamics

Early studies on the outer mantle epithelium (OME) cells of the freshwater bivalve Anodonta cygnea (Linnaeus, 1758) revealed high ionic calcium concentrations by electrophysiological methods and subsequently a high tendency to reach an intracellular toxic condition. This toxicity could be neutralized by specific mechanisms in the cytosol of OME cells of A. cygnea. The present immunocytochemistry studies of OME cells by light and transmission electron microscopy (TEM) clearly showed a positive reaction of an antibody directed against the human plasma membrane Ca(2+)-ATPase 1 (PMCA-1) in the cytoplasm of OME cells. Also, western blot analysis of different fractions of OME cells with anti human PMCA-1 and C28R2 antibodies confirmed the presence of a PMCA-like protein with an unusual topographical localization and a molecular weight of only 70-80 kDa. These results lead us to speculate that this PMCA-like protein is distributed either in the plasma membrane or in the entire cytosol, where it eventually regulates intracellular calcium levels. Interestingly, the antibody reactions showed seasonal variations, being highest in OME samples prepared during summer when A. cygnea live under natural acidosis and absent in samples taken in winter conditions, which is in accordance with the seasonal variation of shell calcification rates. During winter, PMCA-1 antibody reaction was also detected in OME cells of animals kept only under experimentally induced acidosis conditions. Therefore, we assume that a functional role for this PMCA-like protein in the intracellular calcium regulation of OME cells during the mineralization of the shells of A. cygnea can be speculated.

Localization of two genes encoding plasma membrane Ca 2+-transporting ATPases to human chromosomes 1q25–32 and 12q21–23

Human plasma membrane Ca 2+-ATPase (PMCA) isoforms are encoded by at least four separate genes and the diversity of these enzymes is further increased by alternative splicing of transcripts. Cloned cDNAs for two of these isoforms have been used as probes to localize chromosomally the human PMCA1 (ATP2B1) gene to 12q21–q23 and PMCA4 (ATP2B2) to 1q25–q32. These results were obtained by three independent methods, including Southern analysis of human-rodent somatic cell hybrids, in situ hybridization of human metaphase spreads, and genetic linkage analysis in the CEPH pedigrees. High-frequency RFLPs detected at each locus were used in these linkage analyses. No evidence was obtained for multiple copies of the gene at either locus. A cross-hybridizing sequence was detected with PMCA4 probes on Xq13-qter at low stringency. Further studies are required to determine whether this X-chromosomal sequence represents a third member of the PMCA gene family.