Metabolic stress response implicated in diabetic retinopathy: The role of calpain, and the therapeutic impact of calpain inhibitor

Shanab, Ahmed Y.; Nakazawa, Toru; Ryu, Morin; Tanaka, Yuji; Himori, Noriko; Taguchi, Keiko; Yasuda, Masayuki; Watanabe, Ryoji; Takano, Jiro; Saido, Takaomi C.; Minegishi, Naoko; Miyata, Toshio; Abe, Toshiaki; Yamamoto, Masayuki

doi:10.1016/j.nbd.2012.07.025

Cited by 58 publications

(40 citation statements)

References 45 publications

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“…Adult mouse retinal mixed neuronal culture was prepared as previously described [25]. 20 retinas from 6week-old mice were dissected in Hank's buffered-saline solution (HBSS) and incubated at 37°C for 10 min in a digestion solution containing papain (10 U/mL; Worthington, Lakewood, NJ) and L-cysteine (0.3 mg/mL; Sigma) in HBSS.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2013

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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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Section: Methodsmentioning

confidence: 99%

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

et al. 2013

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“…Studies suggested that calpain plays a crucial role in retinal ganglion cell death caused by hyperglycemia and oxidative stress, in which treatment with calpain inhibitor prevented neuronal dysfunction in diabetic retinopathy (Shanab et al, 2012). Recent reports suggested that calpain activation mediates mitochondrial ROS generation and endothelium-dependent dysfunction in diabetes.…”

Section: Discussionmentioning

confidence: 99%

Calpain inhibitor, MDL 28170 confer electrophysiological, nociceptive and biochemical improvement in diabetic neuropathy

2015

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“…Total RNA (200 ng per sample) from the samples was first reverse-transcribed into cDNA using SuperScript III (Invitrogen Life Technologies, Carlsbad, CA). Quantitative RT-PCR was then performed with a 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA) as previously described, with minor modifications [73]. For each 20 μl reaction the following were used: 10 μl TaqMan Fast Universal PCR Master Mix (Applied Biosystems, Foster City, CA), 1 μl Taqman probe, 1 μl template DNA, and 8 μl DEPC water.…”

Section: Methodsmentioning

confidence: 99%

RNA Sequence Reveals Mouse Retinal Transcriptome Changes Early after Axonal Injury

et al. 2014

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Glaucoma is an ocular disease characterized by progressive retinal ganglion cell (RGC) death caused by axonal injury. However, the underlying mechanisms involved in RGC death remain unclear. In this study, we investigated changes in the transcriptome profile following axonal injury in mice (C57BL/6) with RNA sequencing (RNA-seq) technology. The experiment group underwent an optic nerve crush (ONC) procedure to induce axonal injury in the right eye, and the control group underwent a sham procedure. Two days later, we extracted the retinas and performed RNA-seq and a pathway analysis. We identified 177 differentially expressed genes with RNA-seq, notably the endoplasmic reticulum (ER) stress-related genes Atf3, Atf4, Atf5, Chac1, Chop, Egr1 and Trb3, which were significantly upregulated. The pathway analysis revealed that ATF4 was the most significant upstream regulator. The antioxidative response-related genes Hmox1 and Srxn1, as well as the immune response-related genes C1qa, C1qb and C1qc, were also significantly upregulated. To our knowledge, this is the first reported RNA-seq investigation of the retinal transcriptome and molecular pathways in the early stages after axonal injury. Our results indicated that ER stress plays a key role under these conditions. Furthermore, the antioxidative defense and immune responses occurred concurrently in the early stages after axonal injury. We believe that our study will lead to a better understanding of and insight into the molecular mechanisms underlying RGC death after axonal injury.

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Metabolic stress response implicated in diabetic retinopathy: The role of calpain, and the therapeutic impact of calpain inhibitor

Cited by 58 publications

References 45 publications

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

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

Calpain inhibitor, MDL 28170 confer electrophysiological, nociceptive and biochemical improvement in diabetic neuropathy

RNA Sequence Reveals Mouse Retinal Transcriptome Changes Early after Axonal Injury

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