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Expression of dynamin and nitric oxide synthase (NOS) isoforms in rat and mouse collecting ducts
Author(s) -
Hyndman Kelly Anne,
Kim Eujin,
Pollock Jennifer S
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.1025.20
Subject(s) - nos1 , dynamin , gene isoform , microbiology and biotechnology , blot , nitric oxide synthase , biology , knockout mouse , nitric oxide , chemistry , endocytosis , endocrinology , biochemistry , receptor , gene
Dynamins are a family of proteins (DNM1, DNM2 and DNM3) involved in facilitating endocytosis. Recently, a report linked increased dynamin activity to S‐nitrosylation of DNM. We reported that DNM2 and DNM3 interact with NOS1 and NOS3 in the rat kidney inner medulla. The renal inner medulla is comprised of various tubular and vascular structures. The purpose of this study was to determine the expression of NOS and DNM isoforms in isolated rat and mouse renal collecting ducts (CD) and whether DNM is S‐nitrosylated in the CD. Primary suspensions of CDs were prepared by collagenase digestion from rat and mouse inner medulla. CD suspensions and mIMCD3 cells were homogenized and Western blotting analysis for NOS and DNM isoforms performed. Rat CD suspensions express NOS1α, NOS1β, and NOS3; whereas mouse CD suspensions and mIMCD3 express NOS1β and NOS3. Rat CD, mouse CD, and mIMCD3 cells express DNM2 and DNM3. We found little to no expression of NOS2 or DNM1 in any of the CD preparations. Rat CDs were immunoprecipitated with anti‐DNM2 followed with immunoblotting with anti‐S‐nitrosocysteine. We found that DNM2 in rat CD is S‐nitrosylated. In conclusion, these three different models can be used to test questions of DNM/NOS interactions in the CD. This work was funded by NIH HL60653 to JSP; NKF Postdoctoral fellowship to KAH; MCG Diabetes and Obesity Discovery Institute Scholar award to EK.