Unilateral ureteral obstruction results in growth of both the obstructed contralateral kidney. In the obstructed kidney, renal tubular cell trophy and cell death follow an initial brief phase of interstitial proliferation. The contralateral undergoes hypertrophy. To evaluate the active molecular events associated with both renal growth and death in vivo a renal obstruction model will be studied. Use of this model will aid in analyzing the role of specific gene products during both processes and to determine the mechanisms by which quiescent renal cells may be stimulated to either hypertrophy or death in response to obstruction. 1) To determine the role of cell growth genes during renal hypertrophy/death Northern and Western blot analysis of obstructed/unobstructed kidneys will be performed. Concomitantly in situ procedures will be performed for histologic confirmation. 2) Calcium channel alteration will be studied during ureteral obstruction and modified by use of calcium-channel blocking drugs. 3) TRPM-2 is an evolutionarily conserved gene, expressed exclusively in atrophic and dying cells and tissues from a wide variety of species. With the use of recombinant DNA techniques, we will clone, sequence and functionally characterize this gene to determine whether its activation is the cause of cell death or whether it is induced in dying tissues in an attempt to avert this process. In either case, it is a fascinating marker of tissue degeneration and atrophy. Antibodies will be produced in order to determine its cellular localization and other physical characteristics of the TRPM-2 protein product. 1) To determine the role of cell growth genes during renal hypertrophy/death Northern and Western blot analysis of obstructed/unobstructed kidneys will be performed. Concomitantly in situ procedures will be performed for histologic confirmation. 3) Calcium channel alteration will be studied during ureteral obstruction and modified by use of calcium-channel blocking drugs. 3) TRPM-2 is an evolutionarily conserved gene, expressed exclusively in atrophic and dying cells and tissues from a wide variety of species. With the use of recombinant DNA techniques, we will clone, sequence and functionally characterize this gene to determine whether its activation is the cause of cell death or whether it is induced in dying tissues in an attempt to avert this process. In either case, it is a fascinating marker of tissue degeneration and atrophy. Antibodies will be produced in order to determine its cellular localization and other physical characteristics of the TRPM-2 protein product.
