Adeno-associated Virus-mediated Ezh2 Knockdown Reduced the Increment of Newborn Neurons Induced by Forebrain Ischemia in Gerbil Dentate Gyrus

Yoshihide Sehara; Yuki Hashimotodani; Ryota Watano; Kenji Ohba; Ryosuke Uchibori; Kuniko Shimazaki; Kensuke Kawai; Hiroaki Mizukami

doi:10.1007/s12035-024-04200-w

Adeno-associated Virus-mediated Ezh2 Knockdown Reduced the Increment of Newborn Neurons Induced by Forebrain Ischemia in Gerbil Dentate Gyrus

Mol Neurobiol. 2024 Apr 27. doi: 10.1007/s12035-024-04200-w. Online ahead of print.

Authors

Yoshihide Sehara¹, Yuki Hashimotodani², Ryota Watano³, Kenji Ohba³, Ryosuke Uchibori³, Kuniko Shimazaki⁴, Kensuke Kawai⁴, Hiroaki Mizukami³

Affiliations

¹ Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi, 329-0498, Japan. ysehara@jichi.ac.jp.
² Graduate School of Brain Science, Doshisha University, Kyotanabe, Japan.
³ Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi, 329-0498, Japan.
⁴ Department of Neurosurgery, Jichi Medical University, Shimotsuke, Japan.

PMID: 38676810
DOI: 10.1007/s12035-024-04200-w

Abstract

It is established that neurogenesis of dentate gyrus is increased after ischemic insult, although the regulatory mechanisms have not yet been elucidated. In this study, we focused on Ezh2 which suppresses gene expression through catalyzing trimethylation of lysine 27 of histone 3. Male gerbils were injected with adeno-associated virus (AAV) carrying shRNA targeting to Ezh2 into right dentate gyrus 2 weeks prior to forebrain ischemia. One week after ischemia, animals were injected with thymidine analogue to label proliferating cells. Three weeks after ischemia, animals were killed for histological analysis. AAV-mediated knockdown of Ezh2 significantly decreased the ischemia-induced increment of proliferating cells, and the proliferated cells after ischemia showed significantly longer migration from subgranular zone (SGZ), compared to the control group. Furthermore, the number of neural stem cells in SGZ significantly decreased after ischemia with Ezh2 knockdown group. Of note, Ezh2 knockdown did not affect the number of proliferating cells or the migration from SGZ in the non-ischemic condition. Our data showed that, specifically after ischemia, Ezh2 knockdown shifted the balance between self-renewal and differentiation toward differentiation in adult dentate gyrus.

Keywords: Adeno-associated virus vector; Gerbil; Neurogenesis; Polycomb group complex; Transient forebrain ischemia.