Upregulation of cardiac cell plasma membrane calcium pump in a canine model of Chagas disease.
We have previously demonstrated that cardiac myocytes isolated from the hearts of adult dogs develop rapid repetitive cytosolic Ca2+ transients, membrane depolarization, and cell contraction by mobilization of sarcoplasmic reticulum Ca2+ stores when exposed to a soluble factor from the trypomastigotes of Trypanosoma cruzi. These findings led us to investigate the regulatory mechanisms of cytosolic Ca2+ in cardiac tissues from dogs chronically infected with T. cruzi. Expression of the plasma membrane calcium pump (PMCA) RNA and protein was determined by Northern and Western blotting, respectively, followed by densitometric analyses. A 642-bp PMCA 1b complementary DNA probe derived from canine epicardial tissue hybridized to 2 major transcripts (7.3 and 5.3 kb) in canine epicardium. Expression of the dominant transcript (7.3 kb) was 77% greater in cardiac tissues obtained from dogs with chronic T. cruzi infection (140 days after inoculation) in comparison with constitutive expression levels in normal dogs. Monoclonal antibody 5F10, known to recognize all isoforms of the PMCA, was used to detect expression of the PMCA protein in epicardial tissue. Expression of a 142-kDa protein was increased by 58% in the cardiac tissues of infected dogs when compared with those from uninfected dogs. To establish a link between the upregulation of PMCA in dogs chronically infected with Chagas disease and the ventricular-based arrhythmias and myocardial failure that occur during this stage of disease both in dogs and humans, further study will be required.
Barr
S C
SC
Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA. scb6@cornell.edu
Pannabecker
T L
TL
Gilmour
R F
RF
Chandler
J S
JS
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D000252:Calcium-Transporting ATPases / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D027682:Cation Transport Proteins; D002462:Cell Membrane / Q000378:metabolism; D014355:Chagas Disease / Q000378:metabolism*; D018076:DNA, Complementary / Q000737:chemistry; D004195:Disease Models, Animal; D004285:Dogs; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000648:ultrastructure; D053499:Plasma Membrane Calcium-Transporting ATPases; D012333:RNA, Messenger / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014349:Trypanosoma cruzi / Q000235:genetics / Q000276:immunology; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D027682:Cation Transport Proteins; D018076:DNA, Complementary; D012333:RNA, Messenger; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S; D053499:Plasma Membrane Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases