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1 Cell Biology, University of Alabama at Birmingham, Birmingham, AL, USA
2 Medicine, Division of Cardiovascular Disease, University of Alabama at Birmingham, Birmingham, AL, USA
* To whom correspondence should be addressed. E-mail: jchatham{at}uab.edu.
We have previously reported that glucosamine and hyperglycemia attenuate the response of cardiomyocytes to IP3 generating agonists such as Angiotensin II (AngII). This appears to be related to an increase in flux through hexosamine biosynthesis pathway (HBP) and decreased Ca2+ entry into the cells; however, a direct link between HBP and intracellular Ca2+ homeostasis has not been established. Therefore, using neonatal rat ventricular myocytes (NRVM), we investigated the relationship between glucosamine treatment, the concentration of UDP-N-acetylglucosamine (UDP-GlcNAc) an end product of the HBP, and the level of protein O-linked-N-acetylglucosamine (O-GlcNAc) on AngII mediated changes in intracellular free Ca2+-ion concentrations ([Ca2+]i). We found that glucosamine blocked AngII induced [Ca2+]i increase and this was associated with a significant increase in UDP-GlcNAc and O-GlcNAc levels. PUGNAc (O-(2-acetamido-2-deoxy-D-glucopyranosylidene)-amino-N-phenylcarbamate) an inhibitor of O-GlcNAcase, which increased O-GlcNAc levels without changing UDP-GlcNAc concentrations, mimicked the effect of glucosamine of AngII induced increase in [Ca2+]i. An inhibitor of O-GlcNAc-transferase, alloxan prevented the glucosamine-induced increase in O-GlcNAc but not the increase in UDP-GlcNAc; however it abrogated the inhibition of the AngII induced increase in [Ca2+]i. These data support the notion changes in O-GlcNAc levels mediated via increased HBP flux may be involved in the regulation of [Ca2+]i homeostasis in heart.
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