Laboratory Investigation

Kidney International (1991) 40, 997–1006; doi:10.1038/ki.1991.307

Localization and polar distribution of several G-protein subunits along nephron segments

Nigel Brunskill1, Bahar Bastani1, Catalina Hayes1, Jerry Morrissey1 and Saulo Klahr1

1Renal Division, Washington University School of Medicine and the Department of Medicine, Jewish Hospital at Washington University Medical Center, St. Louis, Missouri 63110, USA

Correspondence: Saulo Klahr MD, Department of Medicine, Jewish Hospital at Washington University Medical Center, 216 South Kingshighway, St. Louis, Missouri 63110, USA.

Received 1 April 1991; Revised 8 July 1991; Accepted 11 July 1991.

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Abstract

Localization and polar distribution of several G-protein subunits along nephron segments. Guanine nucleotide binding proteins (G-proteins) are heterotrimeric proteins involved in signal transduction in epithelia. In addition to possessing a polarized epithelium the nephron is composed of well defined segments, each with distinct receptors, transporters and other functions. Since different G-proteins interact with different receptors, a differential distribution of the various G-protein subunits would be expected. To investigate this possibility, polyclonal antipeptide antibodies to several G-protein subunits were used to examine the distribution of these proteins in nephron segments and their localization to apical or basal membrane domains. Immunoblotting of cortical membrane vesicles demonstrated the presence of G-proteins in both the brush border and basolateral membrane. Staining with Galpha common and Galphas antibodies demonstrated more of these G-protein subunits in the brush border than basolateral membrane G-protein beta subunits were also present in greater quantity in brush border membranes Immuno-cytochemical analysis demonstrated good antigenic preservation and basically confirmed the results of immunoblotting of renal cortex G-proteins were also found to be differentially distributed in the medulla, the amounts increasing with proximity to inner medulla. Only Galphas and Gbeta subunits were demonstrated in glomeruli, but all subunits examined were detected in brush borders of proximal and apical membrane of distal tubules in cortex. In outer medulla Galphas was detected in both basolateral and brush borders of some tubules and only brush borders of others, Galphai2 and Gbeta were found in brush border and Galphai3 was not detected in this area. Basolateral membranes of inner medulla contained Galphas, Galphai3, and Gbeta subunits; Galphai3 was also present in apical membranes in some tubules. Galphai2 was detected only in apical membranes in this region. We conclude that G-proteins are differentially distributed in nephron segments. Investigation of regional tubular function may disclose a role for the G-proteins we have localized.

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