TY - JOUR
T1 - Ecel1 knockdown with an AAV2-mediated CRISPR/Cas9 system promotes optic nerve damage-induced RGC death in the mouse retina
AU - Sato, Kota
AU - Shiga, Yukihiro
AU - Nakagawa, Yurika
AU - Fujita, Kosuke
AU - Nishiguchi, Koji
AU - Tawarayama, Hiroshi
AU - Murayama, Namie
AU - Maekawa, Shigeto
AU - Yabana, Takeshi
AU - Omodaka, Kazuko
AU - Katayama, Shota
AU - Feng, Qiwei
AU - Tsuda, Satoru
AU - Nakazawa, Toru
N1 - Funding Information:
The authors thank Yoko Hirata (Gifu University, Japan) for generously providing the mouse HT22 hippocampal cells, Tim Hilts for reviewing and editing the language of the manuscript, Junko Sato, Kanako Sakai, and Eriko Kamii for their technical assistance, and the Biomedical Research Unit of Tohoku University Hospital for technical support. Supported by Grant-in-Aid from the Ministry of Education, Science and Technology of Japan 15K20247 (KS) and 17K16956 (ST). Disclosure: K. Sato, None; Y. Shiga, None; Y. Nakagawa, None; K. Fujita, None; K.M. Nishiguchi, None; H. Tawarayama, None; N. Murayama, None; S. Maekawa, None; T. Yabana, None; K. Omodaka, None; S. Katayama, None; Q. Feng, None; S. Tsuda, None; T. Nakazawa, None
Funding Information:
The authors thank Yoko Hirata (Gifu University, Japan) for generously providing the mouse HT22 hippocampal cells, Tim Hilts for reviewing and editing the language of the manuscript, Junko Sato, Kanako Sakai, and Eriko Kamii for their technical assistance, and the Biomedical Research Unit of Tohoku University Hospital for technical support. Supported by Grant-in-Aid from the Ministry of Education, Science and Technology of Japan 15K20247 (KS) and 17K16956 (ST).
Publisher Copyright:
© 2018 The Authors.
PY - 2018/8
Y1 - 2018/8
N2 - PURPOSE. To assess the therapeutic potential of endothelin-converting enzyme-like 1 (Ecel1) in a mouse model of optic nerve crush. METHODS. Ecel1 expression was evaluated with real time quantitative (qRT)-PCR, Western blotting, and immunohistochemistry in mouse retinas after optic nerve crush. Vinblastine administration to the optic nerve and the intravitreal injection of N-methyl-d-aspartate (NMDA) were used to assess Ecel1 gene expression. Ecel1 was deleted with an adeno-associated viral (AAV) clustered regulatory interspaced short palindromic repeat (CRISPR)/ Cas9 system, and retinal ganglion cell (RGC) survival was investigated with retrograde labeling, qRT-PCR, and visual evoked potential. RESULTS. Optic nerve crush induced Ecel1 expression specifically in the RGCs, peaking on day 4 after optic nerve crush. Ecel1 gene expression was induced by the vinblastine-induced inhibition of axonal flow, but not by NMDA-induced excitotoxicity, even though both are triggers of RGC death. Knockdown of Ecel1 promoted the loss of RGCs after optic nerve crush. CONCLUSIONS. Our data suggest that Ecel1 induction is part of the retinal neuroprotective response to axonal injury in mice. These findings might provide insight into novel therapeutic targets for the attenuation of RGC damage, such as occurs in traumatic optic neuropathy.
AB - PURPOSE. To assess the therapeutic potential of endothelin-converting enzyme-like 1 (Ecel1) in a mouse model of optic nerve crush. METHODS. Ecel1 expression was evaluated with real time quantitative (qRT)-PCR, Western blotting, and immunohistochemistry in mouse retinas after optic nerve crush. Vinblastine administration to the optic nerve and the intravitreal injection of N-methyl-d-aspartate (NMDA) were used to assess Ecel1 gene expression. Ecel1 was deleted with an adeno-associated viral (AAV) clustered regulatory interspaced short palindromic repeat (CRISPR)/ Cas9 system, and retinal ganglion cell (RGC) survival was investigated with retrograde labeling, qRT-PCR, and visual evoked potential. RESULTS. Optic nerve crush induced Ecel1 expression specifically in the RGCs, peaking on day 4 after optic nerve crush. Ecel1 gene expression was induced by the vinblastine-induced inhibition of axonal flow, but not by NMDA-induced excitotoxicity, even though both are triggers of RGC death. Knockdown of Ecel1 promoted the loss of RGCs after optic nerve crush. CONCLUSIONS. Our data suggest that Ecel1 induction is part of the retinal neuroprotective response to axonal injury in mice. These findings might provide insight into novel therapeutic targets for the attenuation of RGC damage, such as occurs in traumatic optic neuropathy.
KW - Ecel1
KW - Neuroprotection
KW - Optic nerve injury
KW - Retinal ganglion cells
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U2 - 10.1167/iovs.18-23784
DO - 10.1167/iovs.18-23784
M3 - Article
C2 - 30073365
AN - SCOPUS:85051198293
VL - 59
SP - 3943
EP - 3951
JO - Investigative Ophthalmology
JF - Investigative Ophthalmology
SN - 0146-0404
IS - 10
ER -