Targeted genomic disruption of H-ras and N-ras has no effect on early renal changes after unilateral ureteral ligation.
OBJECTIVE
To assess the contribution of two different Ras monomeric GTPases isoforms H- and N-Ras in the early changes associated to obstructive nephropathy induced by unilateral ureteral obstruction (UUO).


METHODS
UUO was performed in N-ras (N-ras−/−) and H-ras (H-ras−/−) knock-out mice and control (H-ras+/+/N-ras+/+) mice of C57Bl/6 background. Fibronectin, α-smooth muscle actin, cleaved caspase-3, ki-67, Ras-GTP, pERK, and pAkt expression was analyzed by western blot and/or immunohistochemistry. Ras isoforms activation and caspase activity were determined by both western blot and ELISA.


RESULTS
Three days after UUO, obstructed (O) kidneys of H-ras−/−, N-ras−/−and H-ras+/+/N-ras+/+mice showed no significant differences in activated total ras, pERK1/2, pAkt, total Akt levels, fibronectin, α-SMA expression, cell proliferation, and activated caspase-3. The morphological alterations in the O kidneys, revealed by histological and immunohistochemical studies, were also similar in H-ras−/−, N-ras−/−, and H-ras+/+/N-ras+/+mice.


CONCLUSIONS
These data suggest that the activation of H-ras and N-ras isoforms does not play a major role in the early renal damage induced by UUO.
Grande
María T
MT
Departamento de Fisiología y Farmacología, Instituto Reina Sofía de Investigación Nefrológica, Universidad de Salamanca, Edificio Departamental, Campus Miguel de Unamuno, 37007 Salamanca, Spain.
Arévalo
Miguel
M
Núñez
Alejandro
A
Cannata-Andía
Jorge B
JB
Santos
Eugenio
E
López-Novoa
José M
JM
D000199:Actins / Q000378:metabolism; D000818:Animals; D053148:Caspase 3 / Q000378:metabolism; D004195:Disease Models, Animal; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D016678:Genome; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D007674:Kidney Diseases / Q000473:pathology / Q000503:physiopathology*; D008026:Ligation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics* / Q000378:metabolism*; D014513:Ureter / Q000502:physiology*; D014517:Ureteral Obstruction / Q000473:pathology / Q000503:physiopathology*
D000199:Actins; D005353:Fibronectins; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases; C505409:Casp3 protein, mouse; D053148:Caspase 3; D016283:Proto-Oncogene Proteins p21(ras)