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Articles in PresS, published online ahead of print March 7, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.01028.2001
Submitted on November 29, 2001
Accepted on February 19, 2002
1 Medicine, State University of New York @ Stony Brook, Stony Brook, NY, USA
2 Physiology and Biophysics, State University of New York @ Stony Brook, Stony Brook, NY, USA
3 Pharmacology, New York Medical College, Valhalla, NY, USA
4 Medicine, State University of New York @ Stony Brook, Stony Brook, NY, USA; Physiology and Biophysics, State University of New York @ Stony Brook, Stony Brook, NY, USA
* To whom correspondence should be addressed. E-mail: mgoligorsky{at}mail.som.sunysb.edu.
Hyperhomocysteinemia (HHCy) is an established cause of defective vasorelaxation. Gene expression screening of endothelial cells (HUVEC) treated with Hcy revealed that connexin-43 (Cx43) was upregulated. Expression of Cx43 was increased more than 2-fold in Hcy-treated HUVEC. Gap junctional communication (Lucifer yellow and whole-cell patch-clamp) was not enhanced in Hcy-treated HUVEC. HUVEC expressing chimeric Cx43-green fluorescent protein exhibited it at cell-cell contacts in control, but showed redistribution to the intracellular compartment(s) in Hcy-treated cells. Confocal microscopy of HUVEC stained with anti-Cx43, mitochondrial and endoplasmic reticulum fluorescent markers showed the localization of Cx43 to the plasma membrane of control cells, and its co-localization with the mitochondrial marker in Hcy-treated HUVEC. Studies of isolated mitochondria confirmed overexpression of Cx43 in the mitochondria of Hcy-treated HUVEC. Microdissected renal interlobar arteries, which normally exhibit EDHF-induced vasorelaxation, showed reduced NOS- and COX-independent vasorelaxation to acetylcholine after pretreatment with Hcy. In summary, Hcy-induced upregulation of Cx43 transcript and protein expression, associated with unaltered intercellular communication, redistribution of Cx43 in HUVEC and reduced NO- and prostanoid-independent vascular responses to acetylcholine in Hcy-treated arteries.
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