Diabetes-induced changes of proteins synthesized by adult cardiac myocytes are partially reversed by insulin.
In the rat heart diabetes mellitus leads to a change in myosin heavy chain (MHC) mRNAs and corresponding alterations in myosin isoenzymes as well as a decrease in total cardiac protein synthesis. However, it is still unknown whether cardiac proteins other than MHC are altered by diabetes and if so whether these abnormalities are mediated by insulin deficiency. To answer these questions we analyzed proteins synthesized by isolated cardiac myocytes in the presence or absence of insulin. Enzymatically dispersed adult cardiac myocytes from control and streptozotocin-induced diabetic rats were incubated in medium containing [35S]-methionine for 4 h; diabetic cells were incubated with or without the addition of 5 x 10(-7)M insulin. The labelled peptides were then separated by two-dimensional polyacrylamide gel electrophoresis and analyzed by fluorometry. The abundance of six individual polypeptides was consistently affected by diabetes: one protein was significantly decreased while four others were increased in diabetic myocytes. The remaining protein showed a shift in isoelectric point without a change in molecular weight possibly representing isoforms of a single polypeptide. The addition of insulin reverted the predominance of three proteins back to normal while it did not affect the other three at all. In conclusion diabetes induces changes in the abundance of a few proteins synthesized in vitro by cardiac myocytes and only half of them show an acute response to insulin.
Cagliero
E
E
Department of Medicine, University of California, San Diego.
Barrieux
A
A
Lehman
C
C
Dillmann
W H
WH
D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006321:Heart / Q000187:drug effects; D007328:Insulin / Q000097:blood / Q000494:pharmacology*; D008297:Male; D008715:Methionine / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values; D013462:Sulfur Radioisotopes
D001786:Blood Glucose; D007328:Insulin; D011506:Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine