Differential expression of pkc isoforms in fresh and cultured
rabbit ccd.
Wilborn, Teresa W., and James A. Schafer.
Departments of Physiology and Biophysics, Nephrology Training and
Research Center, University of Alabama at Birmingham, Birmingham, AL
35294
APStracts 2:0201F, 1995.
Growth of rabbit cortical collecting duct (CCD) cells in primary
culture results in a phenotype in which there is a stable stimulation
of Na+ transport by AVP. Our objective was to determine if this
altered phenotype was associated with altered expression of protein
kinase C (PKC) isoforms. Western blot analysis of extracted proteins
and RT/PCR analysis of extracted RNA both showed expression of PKC-a,
e and z in microdissected CCD segments, and in both fresh and
cultured immunodissected CCD cells. These techniques also suggested
that the rabbit CCD expresses PKC-h and q-like isoforms, which have
not as yet been identified in this species. Growth of CCD cells in
primary culture produced no apparent change in the level of
expression of PKC-a, e or z; however, the q-like isoform was strongly
expressed in fresh but only weakly expressed in cultured CCDs. The
putative h isoform was more heavily expressed in cultured than in
fresh CCD cells. The consequences of altered expression of these two
isoforms in fresh and cultured rabbit CCD remains unknown but the
possiblility exists that they are involved in the altered AVP
-response of cultured cells.
Received 12 September 1995; accepted in final form 3 November
1995.
APS Manuscript Number F310-5.
Article publication pending Am. J. Physiol. (Renal Fluid Electrolyte
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 30 November 95