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Neuroprotective and regenerative effect of neurotrophin‐4 on neuronal degeneration induced by advanced glycation end‐products in adult rat retinas
Author(s) -
BIKBOVA G,
OSHITARI T,
YAMAMOTO S
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.4475.x
Subject(s) - dapi , neurite , tunel assay , andrology , retinal , staining , endocrinology , biology , chemistry , medicine , apoptosis , pathology , in vitro , biochemistry
Purpose To determine the effect of AGEs with or without NT‐4 on neuronal cell death and regeneration in isolated rat retinas. Methods Retinal explants of 4 adult SD rats were three‐dimensionally cultured in collagen gel and were incubated either in; 1) serum free control culture media, 2) 10 μg/ml glucose‐AGE‐BSA media, 3) 10 μg/ml glycolaldehyde‐AGE‐BSA media, 4) 10 μg/ml glyceraldehyde‐AGE‐BSA media, 5) glucose‐AGE+100ng/ml NT‐4 media, 6) glycol‐AGE+100ng/ml NT‐4 media, or 7) glycer‐AGE+100 ng/ml NT‐4 supplemented culture media. After 7 days, the number of regenerating neurites from the explants was counted under a phase‐contrast microscope. After counting, retinal explants were fixed, cryosectioned, and stained by TUNEL and DAPI. The ratio of TUNEL‐positive cells to the number of DAPI‐staining nuclei in the ganglion cell layer was calculated. Statistical analysis was performed by one‐way ANOVA. Results In retinas incubated with AGEs (glucose‐AGE, glycol‐AGE, and glycer‐AGE), the numbers of regenerating neurites were fewer than in retinas without AGE (P=0.0033, P=0.0044, and P=0.0238, respectively) and the numbers of TUNEL‐positive cells were higher than in control media (P<0.0001, respectively). NT‐4 supplementation increased, the numbers of regenerating neuritis (P<0.0001) and the numbers of TUNEL‐positives cells were significantly lower than those in glucose‐AGE without NT‐4 (P<0.001) in glycol‐AGE without NT‐4 (P=0.005) and in glycer‐AGE without NT‐4 (P=0.0003). Conclusion AGEs induce neuronal cell death and impede neurite regeneration in adult rat retinas. NT‐4 significantly enhances neuronal survival and regeneration in retinas exposed to AGEs.