Renal distribution of ganglioside GM3 in rat models of types 1 and 2 diabetes
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ORIGINAL PAPER
Renal distribution of ganglioside GM3 in rat models of types 1 and 2 diabetes Anela Novak & Nikolina Režić Mužinić & Vedrana Čikeš Čulić & Joško Božić & Tina Tičinović Kurir & Lejla Ferhatović & Livia Puljak & Anita Markotić
Received: 17 October 2012 / Accepted: 14 March 2013 # University of Navarra 2013
Abstract Ganglioside GM3 is particularly abundant in the kidney tissue and is thought to play an important role in the maintenance of the charge-selective filtration barrier of glomeruli. Altered expression of ganglioside GM3 was pathologically related with glomerular hypertrophy occurring in diabetic human and rat kidneys. Considering the role of GM3 ganglioside in kidney function, the aim of this study was to determine the difference in expression of GM3 ganglioside in glomeruli and tubules using immunofluorescence microscopy both in rat models of types 1 and 2 diabetes mellitus. Diabetes was induced with streptozotocin (55 mg/kg for type 1 diabetes and 35 mg/kg for type 2 diabetes) injection to male Sprague–Dawley rats which were fed with normal pellet diet (type 1 diabetes) or high-fat diet (type 2 diabetes). Rats were sacrificed 2 weeks after diabetes induction, frozen renal sections were stained with primary antibody
GM3(Neu5Ac) and visualized by secondary antibody coupled with Texas red. In addition, renal gangliosides GM3 were analyzed by high-performance thin-layer chromatography followed by GM3 immunostaining. Immunofluorescent microscopy detected 1.7-fold higher GM3 expression in tubules and 1.25-fold higher GM3 in glomeruli of type 1 diabetes mellitus compared with control group. Type 2 diabetes mellitus rats showed slight GM3 increase in whole kidney, unchanged GM3 in glomeruli, but significant higher GM3 expression in tubules, compared with control animals. Taking into consideration increased tubular GM3 content in both types of diabetes, we could hypothesize the role of GM3 in early pathogenesis of diabetic nephropathy.
A. Novak Section of Endocrinology, Department of Internal Medicine, University Hospital Split, Soltanska 1, Split, Croatia
T. Tičinović Kurir (*) Department of Pathophysiology, University of Split School of Medicine, Soltanska 2, 21 000 Split, Croatia e-mail: [email protected]
N. Režić Mužinić : V. Čikeš Čulić : A. Markotić Department of Medical Chemistry and Biochemistry, University of Split School of Medicine, Soltanska 2, Split, Croatia J. Božić Department of Physiology, University of Split School of Medicine, Soltanska 2, Split, Croatia
Keywords Rat . Diabetes mellitus . Streptozotocin . Kidney . GM3
L. Ferhatović : L. Puljak Department of Anatomy, Histology and Embryology, University of Split School of Medicine, Soltanska 2, Split, Croatia
A. Novak et al.
Introduction
Material and methods
Diabetes mellitus is a group of metabolic disorders characterized with hyperglycaemia developed due to dysregulation of insulin excretion and/or action. Diabetes mellitus type 1 is caused in general by the lack of insulin while type 2 diabetes mellitus is
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