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Int. J. Mol. Sci. 2017, 18(12), 2567; doi:10.3390/ijms18122567

Immunohistochemical Characterization of Connexin43 Expression in a Mouse Model of Diabetic Retinopathy and in Human Donor Retinas

1
Buchanan Ocular Therapeutics Unit, Department of Ophthalmology, New Zealand National Eye Centre, University of Auckland, Auckland 1142, New Zealand
2
Department of Ophthalmology, New Zealand National Eye Centre, University of Auckland, Auckland 1142, New Zealand
3
School of Optometry and Vision Science, New Zealand National Eye Centre, University of Auckland, Auckland 1142, New Zealand
4
Centre for Ophthalmology and Visual Science, University of Western Australia, Perth 6009, Western Australia, Australia
*
Author to whom correspondence should be addressed.
Received: 20 October 2017 / Revised: 21 November 2017 / Accepted: 24 November 2017 / Published: 29 November 2017
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Abstract

Diabetic retinopathy (DR) develops due to hyperglycemia and inflammation-induced vascular disruptions in the retina with connexin43 expression patterns in the disease still debated. Here, the effects of hyperglycemia and inflammation on connexin43 expression in vitro in a mouse model of DR and in human donor tissues were evaluated. Primary human retinal microvascular endothelial cells (hRMECs) were exposed to high glucose (HG; 25 mM) or pro-inflammatory cytokines IL-1β and TNF-α (10 ng/mL each) or both before assessing connexin43 expression. Additionally, connexin43, glial fibrillary acidic protein (GFAP), and plasmalemma vesicular associated protein (PLVAP) were labeled in wild-type (C57BL/6), Akita (diabetic), and Akimba (DR) mouse retinas. Finally, connexin43 and GFAP expression in donor retinas with confirmed DR was compared to age-matched controls. Co-application of HG and cytokines increased connexin43 expression in hRMECs in line with results seen in mice, with no significant difference in connexin43 or GFAP expression in Akita but higher expression in Akimba compared to wild-type mice. On PLVAP-positive vessels, connexin43 was higher in Akimba but unchanged in Akita compared to wild-type mice. Connexin43 expression appeared higher in donor retinas with confirmed DR compared to age-matched controls, similar to the distribution seen in Akimba mice and correlating with the in vitro results. Although connexin43 expression seems reduced in diabetes, hyperglycemia and inflammation present in the pathology of DR seem to increase connexin43 expression, suggesting a causal role of connexin43 channels in the disease progression. View Full-Text
Keywords: diabetic retinopathy; connexins; hyperglycemia; inflammation; Akimba diabetic retinopathy; connexins; hyperglycemia; inflammation; Akimba
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MDPI and ACS Style

Mugisho, O.O.; Green, C.R.; Zhang, J.; Binz, N.; Acosta, M.L.; Rakoczy, E.; Rupenthal, I.D. Immunohistochemical Characterization of Connexin43 Expression in a Mouse Model of Diabetic Retinopathy and in Human Donor Retinas. Int. J. Mol. Sci. 2017, 18, 2567.

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