Fluorescence, birefringence and confocal microscopy of the abdominal aorta from nonobese diabetic (NOD) mice.
In this study, nonenzymatic glycosylation was assessed in aorta extracellular matrix (ECM) from nonobese diabetic (NOD) mice, using nitroblue tetrazolium (NBT). Molecular and structural changes were investigated in elastic lamellae and collagen fibers of diabetic mice aortas after staining with dansyl chloride and anilinonaphthalene sulfonate (ANS). Alterations in arterial autofluorescence and birefringence of collagen fibers were investigated in unstained aortas. Proliferation of smooth muscle cells (SMC) was also investigated by Feulgen reaction staining assessed by confocal microscopy and image analysis. Assessment of nonenzymatic glycosylation demonstrated glycosylation products in the aorta ECM of NOD mice. Elastic lamellae and collagen fibers from NOD mouse aortas presented less intense fluorescence after staining with dansyl chloride and ANS when compared to aortas of control nondiabetic mice. However, unstained NOD aortas showed more intense autofluorescence when compared to controls. Birefringence analysis suggests alterations in the higher molecular packing of the arterial collagen fibers in NOD aortas. In aortas stained by Feulgen reaction, no evidence of SMC proliferation was observed in diabetic aortas.
Aldrovani
Marcela
M
Department of Cell Biology, Institute of Biology, State University of Campinas, Cidade Universitária Zeferino Vaz, 13.083-970 Campinas, SP, Brazil. m_ald@unicamp.br
Aparecida Guaraldo
Ana Maria
AM
de Campos Vidal
Benedicto
B
D000818:Animals; D001012:Aorta, Abdominal / Q000378:metabolism / Q000473:pathology*; D001718:Birefringence; D049109:Cell Proliferation; D003094:Collagen / Q000737:chemistry / Q000648:ultrastructure; D003619:Dansyl Compounds / Q000737:chemistry; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism / Q000473:pathology*; D004547:Elastic Tissue / Q000737:chemistry / Q000473:pathology; D006031:Glycosylation; D007202:Indicators and Reagents / Q000737:chemistry; D051379:Mice; D016688:Mice, Inbred NOD; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009131:Muscle, Smooth, Vascular / Q000473:pathology; D009580:Nitroblue Tetrazolium / Q000737:chemistry; D013194:Staining and Labeling
D003619:Dansyl Compounds; D007202:Indicators and Reagents; D009580:Nitroblue Tetrazolium; D003094:Collagen; C029038:dansyl chloride