Diabetes and Overexpression of proNGF Cause Retinal Neurodegeneration via Activation of RhoA Pathway

Mohammed M.H. Al-Gayyar, Barbara A Mysona, Suraporn Matragoon, Mohammed A. Abdelsaid, Mona F. El-Azab, Ahmed Y. Shanab, Yonju Ha, Sylvia B Smith, Kathryn Elizabeth Bollinger, Azza B. El-Remessy

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Abstract

Our previous studies showed positive correlation between accumulation of proNGF, activation of RhoA and neuronal death in diabetic models. Here, we examined the neuroprotective effects of selective inhibition of RhoA kinase in the diabetic rat retina and in a model that stably overexpressed the cleavage-resistance proNGF plasmid in the retina. Male Sprague-Dawley rats were rendered diabetic using streptozotosin or stably express cleavage-resistant proNGF plasmid. The neuroprotective effects of the intravitreal injection of RhoA kinase inhibitor Y27632 were examined in vivo. Effects of proNGF were examined in freshly isolated primary retinal ganglion cell (RGC) cultures and RGC-5 cell line. Retinal neurodegeneration was assessed by counting TUNEL-positive and Brn-3a positive retinal ganglion cells. Expression of proNGF, p75NTR, cleaved-PARP, caspase-3 and p38MAPK/JNK were examined by Western-blot. Activation of RhoA was assessed by pull-down assay and G-LISA. Diabetes and overexpression of proNGF resulted in retinal neurodegeneration as indicated by 9- and 6-fold increase in TUNEL-positive cells, respectively. In vitro, proNGF induced 5-fold cell death in RGC-5 cell line, and it induced >10-fold cell death in primary RGC cultures. These effects were associated with significant upregulation of p75NTR and activation of RhoA. While proNGF induced TNF-α expression in vivo, it selectively activated RhoA in primary RGC cultures and RGC-5 cell line. Inhibiting RhoA kinase with Y27632 significantly reduced diabetes- and proNGF-induced activation of proapoptotic p38MAPK/JNK, expression of cleaved-PARP and caspase-3 and prevented retinal neurodegeneration in vivo and in vitro. Taken together, these results provide compelling evidence for a causal role of proNGF in diabetes-induced retinal neurodegeneration through enhancing p75NTR expression and direct activation of RhoA and p38MAPK/JNK apoptotic pathways.

Original languageEnglish (US)
Article numbere54692
JournalPloS one
Volume8
Issue number1
DOIs
StatePublished - Jan 30 2013

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Retinal Ganglion Cells
Medical problems
diabetes
Chemical activation
Cells
Cell culture
Phosphotransferases
Neuroprotective Agents
Cell death
mitogen-activated protein kinase
Caspase 3
neuroprotective effect
phosphotransferases (kinases)
Rats
cell culture
Cell Culture Techniques
Plasmids
cell lines
In Situ Nick-End Labeling
caspase-3

ASJC Scopus subject areas

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

Cite this

Al-Gayyar, M. M. H., Mysona, B. A., Matragoon, S., Abdelsaid, M. A., El-Azab, M. F., Shanab, A. Y., ... El-Remessy, A. B. (2013). Diabetes and Overexpression of proNGF Cause Retinal Neurodegeneration via Activation of RhoA Pathway. PloS one, 8(1), [e54692]. https://doi.org/10.1371/journal.pone.0054692

Diabetes and Overexpression of proNGF Cause Retinal Neurodegeneration via Activation of RhoA Pathway. / Al-Gayyar, Mohammed M.H.; Mysona, Barbara A; Matragoon, Suraporn; Abdelsaid, Mohammed A.; El-Azab, Mona F.; Shanab, Ahmed Y.; Ha, Yonju; Smith, Sylvia B; Bollinger, Kathryn Elizabeth; El-Remessy, Azza B.

In: PloS one, Vol. 8, No. 1, e54692, 30.01.2013.

Research output: Contribution to journalArticle

Al-Gayyar, MMH, Mysona, BA, Matragoon, S, Abdelsaid, MA, El-Azab, MF, Shanab, AY, Ha, Y, Smith, SB, Bollinger, KE & El-Remessy, AB 2013, 'Diabetes and Overexpression of proNGF Cause Retinal Neurodegeneration via Activation of RhoA Pathway', PloS one, vol. 8, no. 1, e54692. https://doi.org/10.1371/journal.pone.0054692
Al-Gayyar, Mohammed M.H. ; Mysona, Barbara A ; Matragoon, Suraporn ; Abdelsaid, Mohammed A. ; El-Azab, Mona F. ; Shanab, Ahmed Y. ; Ha, Yonju ; Smith, Sylvia B ; Bollinger, Kathryn Elizabeth ; El-Remessy, Azza B. / Diabetes and Overexpression of proNGF Cause Retinal Neurodegeneration via Activation of RhoA Pathway. In: PloS one. 2013 ; Vol. 8, No. 1.
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AU - Abdelsaid, Mohammed A.

AU - El-Azab, Mona F.

AU - Shanab, Ahmed Y.

AU - Ha, Yonju

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AU - Bollinger, Kathryn Elizabeth

AU - El-Remessy, Azza B.

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N2 - Our previous studies showed positive correlation between accumulation of proNGF, activation of RhoA and neuronal death in diabetic models. Here, we examined the neuroprotective effects of selective inhibition of RhoA kinase in the diabetic rat retina and in a model that stably overexpressed the cleavage-resistance proNGF plasmid in the retina. Male Sprague-Dawley rats were rendered diabetic using streptozotosin or stably express cleavage-resistant proNGF plasmid. The neuroprotective effects of the intravitreal injection of RhoA kinase inhibitor Y27632 were examined in vivo. Effects of proNGF were examined in freshly isolated primary retinal ganglion cell (RGC) cultures and RGC-5 cell line. Retinal neurodegeneration was assessed by counting TUNEL-positive and Brn-3a positive retinal ganglion cells. Expression of proNGF, p75NTR, cleaved-PARP, caspase-3 and p38MAPK/JNK were examined by Western-blot. Activation of RhoA was assessed by pull-down assay and G-LISA. Diabetes and overexpression of proNGF resulted in retinal neurodegeneration as indicated by 9- and 6-fold increase in TUNEL-positive cells, respectively. In vitro, proNGF induced 5-fold cell death in RGC-5 cell line, and it induced >10-fold cell death in primary RGC cultures. These effects were associated with significant upregulation of p75NTR and activation of RhoA. While proNGF induced TNF-α expression in vivo, it selectively activated RhoA in primary RGC cultures and RGC-5 cell line. Inhibiting RhoA kinase with Y27632 significantly reduced diabetes- and proNGF-induced activation of proapoptotic p38MAPK/JNK, expression of cleaved-PARP and caspase-3 and prevented retinal neurodegeneration in vivo and in vitro. Taken together, these results provide compelling evidence for a causal role of proNGF in diabetes-induced retinal neurodegeneration through enhancing p75NTR expression and direct activation of RhoA and p38MAPK/JNK apoptotic pathways.

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