Advanced glycation end (AGE) product modification of laminin downregulates Kir4.1 in retinal Müller cells

Kayla Thompson, Jonathan Chen, Qianyi Luo, Yucheng Xiao, Theodore Cummins, Ashay Bhatwadekar

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Diabetic retinopathy (DR) is a major cause of adult blindness. Retinal Müller cells maintain water homeostasis and potassium concentration via inwardly rectifying Kir4.1 channels. Accumulation of advanced glycation end products (AGEs) is a major pathologic event in DR. While diabetes leads to a decrease in the Kir4.1 channels, it remains unknown whether AGEs-linked to the basement membrane (BM) affect normal Kir4.1 channels. For this study, we hypothesized that AGE-modification of laminin is detrimental to Kir4.1 channels, therefore, disrupting Müller cell function. The AGE-modified laminin-coated substrates were prepared by incubating Petri-dishes with laminin and methylglyoxal for seven days. The rat Müller cells (rMC-1) were propagated on AGE-modified laminin, and Kir4.1 expression and function were evaluated. Quantification of AGEs using ELISA revealed a dose-dependent increase in methylglyoxal-hydro-imidazolone adducts. The rMC-1 propagated on AGE-modified laminin demonstrated a decrease in Kir4.1 levels in immunofluorescence and western blot studies and a decrease in the Kir4.1 channel function. Kir4.1 decrease on AGE-modified laminin resulted in a disorganization of an actin cytoskeleton and disruption of α-dystrogly-can-syntrophin-dystrophin complexes. Our studies suggest that AGE-modification of laminin is detrimental to Kir4.1 channels. By studying the role of AGEs in Kir4.1 channels we have identified a novel mechanism of Müller cell dysfunction and its subsequent involvement in DR.

Original languageEnglish (US)
Article numbere0193280
JournalPLoS One
Volume13
Issue number2
DOIs
StatePublished - Feb 1 2018

Fingerprint

Advanced Glycosylation End Products
laminin
Laminin
glycation
Down-Regulation
diabetic retinopathy
Diabetic Retinopathy
Pyruvaldehyde
cells
dystrophin
Dystrophin
blindness
basement membrane
Blindness
Medical problems
advanced glycation end products
microfilaments
Actin Cytoskeleton
cans
Basement Membrane

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Advanced glycation end (AGE) product modification of laminin downregulates Kir4.1 in retinal Müller cells. / Thompson, Kayla; Chen, Jonathan; Luo, Qianyi; Xiao, Yucheng; Cummins, Theodore; Bhatwadekar, Ashay.

In: PLoS One, Vol. 13, No. 2, e0193280, 01.02.2018.

Research output: Contribution to journalArticle

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abstract = "Diabetic retinopathy (DR) is a major cause of adult blindness. Retinal M{\"u}ller cells maintain water homeostasis and potassium concentration via inwardly rectifying Kir4.1 channels. Accumulation of advanced glycation end products (AGEs) is a major pathologic event in DR. While diabetes leads to a decrease in the Kir4.1 channels, it remains unknown whether AGEs-linked to the basement membrane (BM) affect normal Kir4.1 channels. For this study, we hypothesized that AGE-modification of laminin is detrimental to Kir4.1 channels, therefore, disrupting M{\"u}ller cell function. The AGE-modified laminin-coated substrates were prepared by incubating Petri-dishes with laminin and methylglyoxal for seven days. The rat M{\"u}ller cells (rMC-1) were propagated on AGE-modified laminin, and Kir4.1 expression and function were evaluated. Quantification of AGEs using ELISA revealed a dose-dependent increase in methylglyoxal-hydro-imidazolone adducts. The rMC-1 propagated on AGE-modified laminin demonstrated a decrease in Kir4.1 levels in immunofluorescence and western blot studies and a decrease in the Kir4.1 channel function. Kir4.1 decrease on AGE-modified laminin resulted in a disorganization of an actin cytoskeleton and disruption of α-dystrogly-can-syntrophin-dystrophin complexes. Our studies suggest that AGE-modification of laminin is detrimental to Kir4.1 channels. By studying the role of AGEs in Kir4.1 channels we have identified a novel mechanism of M{\"u}ller cell dysfunction and its subsequent involvement in DR.",
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