Immunohistochemical detection of advanced glycosylation end products in diabetic tissues using monoclonal antibody to pyrraline.

Abstract

Pyrraline is one of the major Maillard compounds resulting from the reaction of glucose with amino compounds at slightly acidic pH. For in vivo studies, monoclonal pyrraline antibodies were raised after immunization of Balb/c mice with keyhole limpet hemocyamin-caproyl pyrraline conjugate. Of 660 hybridoma clones from one donor, 260 produced an antibody to the free hapten, two of which named Pyr-A and Pyr-B also cross-reacted with L-lysyl pyrraline. Using Pyr-B antibody and an ELISA, a gradual increase in pyrraline immunoreactivity was observed in serum albumin incubated with glucose or 3-deoxyglucosone. Plasma pyrraline levels increased fourfold (P < 0.001) in Sprague-Dawley rats upon induction of diabetes with streptozotocin and were twofold increased in randomly selected plasmas from diabetic humans. Highly specific pyrraline immunoreactivity was detected in sclerosed glomeruli from diabetic and old normal kidneys as well as in renal arteries with arteriolosclerosis and in perivascular and peritubular sclerosed extracellular matrix and basement membranes. The preferential localization of pyrraline immunoreactivity in the extracellular matrix strengthens the notion that the advanced glycosylation reaction may contribute to decreased turnover and thickening of the extracellular matrix in diabetes and aging. (J. Clin.

Nonenzymatic Glycation at the N Terminus of Pathogenic Prion Protein in Transmissible Spongiform Encephalopathies

“…Furthermore, the deleterious effects of AGEs may be associated with oxidative stress (7)(8)(9)(10)(11)(12). It has been shown that tau, a protein associated with paired helical filaments (PHFs), which plays an important role in AD pathology, is advanced-glycated; AGE-positive immunostaining is present in the neuritic plaques and the neurofibrillary tangles (NFT) in AD brains (13).…”

Accumulation of Advanced Glycation Endproducts in the Rat Nephron: Link with Circulating AGEs During Aging

“…However, there is a link between the pattern of glomerular changes that occur in aging kidneys and the age-dependent accumulation of advanced glycosylation endproducts (AGEs) (Sell and Monnier 1990;Makita et al 1992). AGEs are the late products of the glycation of matrix proteins exposed to glucose and they accumulate slowly in the renal and extrarenal matrices during normal aging Miyata and Monnier 1992) and more rapidly in diabetes as a result of hyperglycemia (Brownlee et al 1988;Makita et al 1992;Vlassara et al 1994a). Glucose tolerance often becomes impaired with aging (Defronzo 1979;Fink et al 1984), and although hyperglycemia is not directly involved in aging, several lines of evidence indicate that the accumulation of GECM in aging kidneys is due to the formation of AGE (Miyata and Monnier 1992;Li et al 1996).…”

“…AGEs are the late products of the glycation of matrix proteins exposed to glucose and they accumulate slowly in the renal and extrarenal matrices during normal aging Miyata and Monnier 1992) and more rapidly in diabetes as a result of hyperglycemia (Brownlee et al 1988;Makita et al 1992;Vlassara et al 1994a). Glucose tolerance often becomes impaired with aging (Defronzo 1979;Fink et al 1984), and although hyperglycemia is not directly involved in aging, several lines of evidence indicate that the accumulation of GECM in aging kidneys is due to the formation of AGE (Miyata and Monnier 1992;Li et al 1996). Glycated basement membrane macromolecules, especially Type IV collagen (Cohen and Wu 1981;Ziyadeh 1993), can affect renal function by altering the permselectivity and resistance to degradation of thickened GBM, which is the hallmark of a glomerular defect.…”

Maillard Reaction Products from 3-Deoxyglucosone and Butylamine under Physiological Conditions.

“…N-Butylacetamide, N-butylformamide, and butylpyrraline were also identified in butylamine-glucose reaction system , suggesting that 3DG is an important intermediate of these products. Lysyl-pyrraline has been reported to increase in samples in vivo with diabetes and aging (Hayase et al, 1989;Miyata & Monnier, 1992). We found that N-butylacetamide and N-butylformamide were Table 1. at pH 7.4 and 50'C.…”