Determination of caveolin-1 in renal caveolar and non-caveolar fractions in experimental type 1 diabetes

Title:

Determination of caveolin-1 in renal caveolar and non-caveolar fractions in experimental type 1 diabetes

Creator:

Hana Demová and Radko Komers

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:788fa988-819f-4ad9-913b-8bfe83bf1023
uuid:788fa988-819f-4ad9-913b-8bfe83bf1023

Subject:

Patologie. Klinická medicína, diabetologie, diabetes mellitus, nemoci ledvin, inzulin, diabetology, kidney diseases, insulin, caveolin-1, type 1 diabetes mellitus, sucrose fractination, losartan, 14, and 616

Type:

article, články, model:article, and TEXT

Description:

1a_Caveolin-1 (CAV-1) is the main structural component of caveolae, acting as a modulator of signal transduction. CAV-1 might be involved in the pathophysiology of microvascular complications in Type 1 diabetes (DM). We sought to determine whether fractionation on sucrose gradient (SF), a method routinely utilized for isolation of caveolar fractions in homogenous cell lines, is applicable for CAV-1-related studies in tissues with multiple cell types, such as the normal rat kidney cortex (C). Using this method, we also determined whether streptozotocin- induced DM in rats (4-week duration) leads to changes in renal subcellular targeting of CAV-1, and evaluated the effects of tight metabolic control (insulin, 12 IU /day) and angiotensin receptor blocker, losartan (4 weeks, 20 mg/kg/day). Immunoblotting of individual fractions obtained from C revealed CAV-1 expression in fractions 4-6 that corresponded to light scattering band that typically forms after separating cellular fractions on SF. These fractions were considered to be caveolar fractions. In C, CAV-1 was also detectable in fracti ons 8-10. These and all other fractions except caveolar fractions were considered to be non-caveolar fractions. A ratio of caveolar/non-caveolar expression of CAV-1 (CNCR) was computed for each renal cortex allowing comparisons of CAV-1 subcellular distribution in C and DM rats, and effects of treatments., 2a_Using this approach, DM was characterized by marked increases in CNCR as compared to C (5.54±1.56 vs. 2.65±1.33, p<0. 05) that were reduced by treatment with insulin (0.78±0. 24, p<0.01 vs. DM) or losartan (0.84±0.06, p<0.01 vs. DM). In summary, analysis of CAV-1 following the SF of renal cortex detected similar distribution of the protein as in homogenous cell lines, DM-induced changes in CAV-1 targeting, and the effects of pharmacological treatments. This suggests applicability of SF in studies focusing on CAV-1 targeting in organs with various cell lines in vivo., H. Demová, R. Komers., and Obsahuje seznam literatury

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 2009 Volume:58 | Number:4

Harvested from:

CDK

Metadata only:

false