adrenomedullin and Abnormalities--Multiple

To investigate the expression of endothelin-1 (ET-1) and adrenomedullin (ADM) in the renal pelvis, stenotic ureteropelvic junction, and ureter of 20 male Wistar rats with congenital unilateral ureteropelvic junction obstruction; the normal contralateral kidneys served as controls. The molecular pathophysiology of congenital ureteropelvic junction obstruction is still unclear. The implication of altered peptidergic innervation is under discussion. Our study group has recently been able to demonstrate a significant increase in ET-1 and a significant decrease in ADM in prestenotic and stenotic tissue, but not in the remainder of the ureter, compared with controls.. Twenty animals were killed, and samples of the renal pelvis, ureteropelvic junction, upper ureter, middle part of the ureter, and lower ureter were immediately snap-frozen and stored in liquid nitrogen. Total RNA was extracted, and subsequently 1 microg of RNA was reversely transcribed. mRNA expression of ET-1 and ADM was determined semiquantitatively using on-line polymerase chain reaction. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was determined to relate the specific mRNA expression to the expression of a housekeeping gene.. We found a significant increase in the expression of ET-1 in the obstructed junctions related to GAPDH (P <0.001). The expression of ADM, however, revealed no statistically significant differences. No differences at all could be detected in the tissue samples from the rest of the ureter.. Alterations in the local production of peptidergic neurotransmitters, especially ET-1, may contribute to the molecular pathogenesis of ureteropelvic junction obstruction. Results previously obtained in the stenotic tissue from children were confirmed in the stenotic tissue from the rat model. We hypothesize that the alterations are disease-, but not age-specific.

Topics: Abnormalities, Multiple; Adrenomedullin; Animals; Atrophy; Computer Systems; Constriction, Pathologic; Disease Models, Animal; Endothelin-1; Gene Expression Profiling; Hydronephrosis; Kidney Pelvis; Male; Peptides; Polymerase Chain Reaction; Rats; Rats, Mutant Strains; Rats, Wistar; RNA, Messenger; Ureter; Ureteral Obstruction

Adrenomedullin, a recently identified potent vasodilator, is expressed widely and has been suggested to have functions ranging from reproduction to blood pressure regulation. To elucidate these functions and define more precisely sites of Adm expression, we replaced the coding region of the Adm gene in mice with a sequence encoding enhanced green fluorescent protein while leaving the Adm promoter intact. We find that Adm(-/-) embryos die at midgestation with extreme hydrops fetalis and cardiovascular abnormalities, including overdeveloped ventricular trabeculae and underdeveloped arterial walls. These data suggest that genetically determined absence of Adm may be one cause of nonimmune hydrops fetalis in humans.

Topics: Abnormalities, Multiple; Adrenomedullin; Animals; Aorta; Carotid Arteries; Chimera; DNA, Complementary; Female; Fetal Death; Fetal Heart; Gene Expression Regulation, Developmental; Gene Targeting; Genes, Reporter; Genotype; Gestational Age; Green Fluorescent Proteins; Heart Ventricles; Hydrops Fetalis; Luminescent Proteins; Male; Mice; Mice, Knockout; Peptides; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction