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(Received for publication, March 4, 1996, and in revised form, May 2, 1996)
From the Laboratoire de Biologie Intégrée des Cellules
Rénales, Département de Biologie Cellulaire et
Moléculaire, CEA Saclay 91191 Gif sur Yvette Cedex, France
and ¶ Laboratoire de Physiologie Cellulaire, Collège de
France, 75231 Paris Cedex 05, France
Expression of Ca2+-inhibitable types
V and VI adenylyl cyclases was studied by reverse
transcription-polymerase chain reaction in rat renal glomeruli and
nephron segments isolated by microdissection. Quantitation of each
mRNA was achieved using a mutant cRNA which differed from the wild
type by substituting two bases to create a new restriction site in the
corresponding cDNA. Type VI mRNA was present all along the
nephron but was more abundant in distal than in proximal segments. The
expression of type V mRNA was restricted to the glomerulus and to
the initial portions of the collecting duct. Expression of the
Ca2+-insensitive type IV mRNA studied on the same
samples was evidenced only in the glomerulus. The functional relevance
of the expression of Ca2+-inhibitable isoforms was studied
by measuring cAMP content in the microdissected outer medullary
collecting duct which expressed both type V mRNA (2367 ± 178 molecules/mm tubular length; n = 8) and type VI
mRNA (5658 ± 543 molecules/mm, n = 8).
Agents known to increase intracellular Ca2+ in this segment
induced a Ca2+-dependent inhibition on either
arginine vasopressin- or glucagon-stimulated cAMP level. The
characteristics of these inhibitions suggest a functional and
differential expression of types V and VI adenylyl cyclases in two
different cell types of the rat outer medullary collecting duct.
Volume 271, Number 32,
Issue of August 9, 1996
pp. 19264-19271
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
FUNCTIONAL CONSEQUENCES FOR REGULATION OF THE cAMP
CONTENT
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