Increased neuronal nitric oxide synthase activity in retinal neurons in early diabetic retinopathy.
OBJECTIVE
There are increased levels of nitric oxide (NO) in diabetic retinas. The purpose of this study was to determine the extent that neuronal nitric oxide synthase (nNOS) contributes to the increased levels of retinal NO in early diabetic retinopathy by examining the expression and activity of nNOS in retinal neurons after 5 weeks of diabetes.


METHODS
Changes in NO levels were measured using NO imaging of retinal neurons in mice with streptozotocin-induced diabetes for five weeks. NO imaging was compared to nNOS localization using immunocytochemistry, and nNOS message and protein levels were measured using quantitative real-time PCR and western blots.


RESULTS
There was a close anatomic correlation between the localization of the increased NO production and the nNOS immunoreactivity in the retinal plexiform layers of diabetic retinas. There was no change in nNOS message, but nNOS protein was decreased and its subcellular localization was altered. Treatment with insulin or aminoguanidine partially ameliorated the increase in NO in diabetic retinas.


CONCLUSIONS
These results suggest that increased nNOS activity is responsible for the majority of increased NO in retinal neurons in early diabetic retinopathy. This supports a role for increased nNOS activity in the early neuronal dysfunction in the diabetic retina.
Giove
Thomas J
TJ
Laboratory of Visual Neurobiology, Department of Biology, Boston University, Boston, MA 02215, USA.
Deshpande
Monika M
MM
Gagen
Christine S
CS
Eldred
William D
WD
D000818:Animals; D003930:Diabetic Retinopathy / Q000201:enzymology* / Q000473:pathology; D005453:Fluorescence; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006146:Guanidines / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012160:Retina / Q000473:pathology; D055351:Retinal Neurons / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology*; D013311:Streptozocin; D013997:Time Factors
D006146:Guanidines; D007328:Insulin; D012333:RNA, Messenger; D009569:Nitric Oxide; D013311:Streptozocin; D052248:Nitric Oxide Synthase Type I; C004479:pimagedine; D019331:NG-Nitroarginine Methyl Ester