Ultrastructural and functional analyses of nephropathy in calmodulin-induced diabetic transgenic mice.
Previous animal models of diabetic nephropathy have used diabetic animals for which the underlying defect was either uncertain or the diabetes was induced by potentially specific toxins. In this report, we describe the renal abnormalities in a transgenic mouse model that develops early-onset diabetes due to overexpression of calmodulin in pancreatic beta cells.



Renal tissues were collected from normal and transgenic mice at 112, 182, and 300 days. These were prepared for light microscopic observation, stained with polyethylenimine (for anionic sites), or rendered acellular by detergent extraction prior to observation by transmission and scanning electron microscopy. Morphometric analysis of glomerular basement membrane thickness was carried out by the "orthogonal intercept" method. Twelve-hour urine samples of fed and fasting mice were collected for urine volume and glucose and protein analyses. Blood glucose, blood urea nitrogen, serum insulin, and creatinine were determined in 60-90-day-old and 255-day-old mice by established methods.



Morphometric analyses revealed age-related and transgene-related increases in glomerular basement membrane thickness. A 22% increase in transgenic diabetics over controls was seen at 112 days of age that developed to increases of 43% and 37% at 182 and 300 days of age, respectively. Mesangial matrix area was also increased markedly in transgenic mice. Surprisingly, even in the oldest diabetic mice, there was no reduction in anionic sites. Moreover, despite an eightfold increase in urine volume, these mice did not become significantly proteinuric.



These results indicate that proteinuria of diabetes may be delayed or prevented by maintenance of a normal complement of glomerular basement membrane anionic sites. They also demonstrate that transgenic mice can provide a valuable model for discriminating between different aspects of diabetic nephropathy.
Carlson
E C
EC
Department of Anatomy, University of North Dakota School of Medicine, Grand Forks, North Dakota 58202, USA.
Audette
J L
JL
Klevay
L M
LM
Nguyen
H
H
Epstein
P N
PN
D000818:Animals; D000838:Anions / Q000737:chemistry; D001485:Basement Membrane / Q000378:metabolism / Q000648:ultrastructure; D001774:Blood Chemical Analysis; D003924:Diabetes Mellitus, Type 2 / Q000209:etiology*; D005109:Extracellular Matrix / Q000473:pathology; D007668:Kidney / Q000502:physiology / Q000648:ultrastructure*; D007674:Kidney Diseases / Q000473:pathology* / Q000503:physiopathology; D007678:Kidney Glomerulus / Q000473:pathology; D051379:Mice; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D016482:Urinalysis
D000838:Anions