Ethanol ingestion induces Shh and Gli1 production and accumulation in renal pericytes.

A) Renal Shh and PDGFR-ß1 co-localize after ethanol ingestion. Fluorescent immunohistochemistry of PDGFR-ß1, Shh, or DAPI nuclear counterstain of renal sections from mice ingesting a ramped ethanol diet or pair-fed a control diet. Image overlay in the right panel of green PDGFR-β and red Shh shows co-expression in yellow. B) Ethanol induces Gli1 transcription. ß-Galactosidase expression in kidney under control of the gli1 locus in kidneys of pair-fed control and ethanol-fed heterozygotic Gli1tm2Alj/J reporter mice was assessed by hydrolysis of fluorogenic LacZ as described in “Methods.” C) Collagen accumulates in the kidneys of wild-type and Gli1 reporter mice. Immunoblots of collagen 1 protein in kidney lysates of wild-type or gli1-LacZ mice pair-fed a control diet or fed the ramped ethanol diet for 25 days. D) Collagen message is increased in Gli1 reporter mice. Collagen 1 mRNA quantified by qPCR relative to ribosomal 18s RNA in the kidneys of wild-type or reporter mice. SEM (n = 4), * p<0.05 E) Ethanol feeding damages renal function in Gli1 reporter mice. Circulating creatinine concentrations in gli1-LacZ or wild-type mice ingesting ethanol or a paired control diet with isomaltose isocalorically substituting for ethanol. SEM (n = 4). *, p<0.05