P2X7 subunit-like immunoreactivity in the nucleus of visceral smooth muscle cells of the guinea pig.
P2X receptors are adenosine 5'-triphosphate (ATP)-gated cation-selective channels. Seven cloned subunits (P2X(1-7)) are found in the plasma membrane of many cells in the brain and periphery. ATP-dependent opening of nuclear membrane channels, which resemble P2X(7) receptors, has been reported previously. Therefore, the aim of this study was to use specific antisera to examine P2X(7) receptor subunit expression in the nucleus of cells in visceral organs. In sections of the guinea pig vas deferens and bladder, widespread nuclear immunoreactivity was observed in smooth muscle, epithelial, and lamina propria cells using two antisera raised against different sequences within the P2X(7) subunit. Nuclear immunostaining was also seen in a variety of cell types in the myocardium, lung, stomach, ileum, and colon of the guinea pig. In all tissues, omission of the primary antisera or preincubation of the primary antisera with the antigenic peptide resulted in the loss of specific immunostaining, as did dilution of the antisera beyond 1:32000 or omitting antigen retrieval. Western blot analysis revealed that the anti-P2X(7) antiserum recognised a protein in membrane preparations from both guinea pig vas deferens nuclei and HEK-293 cells transfected with P2X(7) subunits. Thus, the nuclear expression of P2X(7) receptors appears to be widespread in smooth muscle and epithelial cells of visceral organs.
Menzies
John
J
Department of Physiology and Pharmacology, University of Strathclyde, Strathclyde Institute for Biomedical Sciences, John Arbuthnott Building, 27, Taylor Street, Glasgow, G4 0NR, UK. jmenzies@dacc.uchicago.edu
Paul
Andrew
A
Kennedy
Charles
C
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D006224:Cricetinae; D006168:Guinea Pigs; D006801:Humans; D019044:Image Cytometry; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018048:Receptors, Purinergic P2 / Q000378:metabolism*; D058486:Receptors, Purinergic P2X7; D013045:Species Specificity; D014162:Transfection / Q000379:methods; D001743:Urinary Bladder / Q000166:cytology*
C546468:P2RX7 protein, human; D018048:Receptors, Purinergic P2; D058486:Receptors, Purinergic P2X7