中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (4): 578-590.doi: 10.4103/1673-5374.180742

• 原著:视神经损伤修复保护与再生 • 上一篇    下一篇

成年虹鳟鱼损伤视神经和大脑整合中枢神经再生过程中神经细胞增殖与凋亡

  

  • 收稿日期:2015-12-22 出版日期:2016-04-30 发布日期:2016-04-30
  • 基金资助:

    俄罗斯总统基金；俄罗斯科学科学学会基金；印度政府科技部DST-INSPIRE基金

Cell proliferation and apoptosis in optic nerve and brain integration centers of adult trout Oncorhynchus mykiss after optic nerve injury

Evgeniya V. Pushchina1, *, Sachin Shukla2, Anatoly A. Varaksin1, Dmitry K. Obukhov3   

  1. 1 Laboratory of Cytophysiology, A.V. Zhirmunsky Institute of Marine Biology Far Eastern Branch of Russian Academy of Sciences, Vladivostok, Russia
    2 Prof. Brien Holden Eye Research Centre, L.V. Prasad Eye Institute, Hyderabad, India
    3 St. Petersburg State University, Universitetskaya nab. 7/9, St. Petersburg, Russia
  • Received:2015-12-22 Online:2016-04-30 Published:2016-04-30
  • Contact: Evgeniya V. Pushchina, D.Sc., Ph.D., puschina@mail.ru.
  • Supported by:

    This work was supported by a grant from President of Russian Federation (No. MD-4318.2015.4), a grant from Program for Basic Research of the Far East Branch of the Russian Academy of Sciences 2015–2017 (No. 15-I-6-116, section III), and DST-INSPIRE Faculty Grant (No. IFA14-LSBM-104) from the Department of Science and Technology (DST), Government of India.

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摘要：

研究已知，鱼在中枢神经损伤后，有重建神经细胞和神经纤维的能力。然而与成年大脑相关的神经发生过程尚不明确。实验应用增殖细胞核抗原免疫组化染色检测成年虹鳟鱼视神经和脑损伤后神经细胞增殖。TUNEL染色检测视神经损伤后神经细胞凋亡。发现视神经损伤后1周，巨噬细胞和小胶质细胞聚集区细胞大量死亡。脑损伤后1周，视顶盖区发现增殖细胞核抗原阳性的放射状胶质细胞样细胞。小脑背侧增殖区和分子颗粒层检测到增殖细胞，而在顶盖和小脑则发现未分化的细胞形成的神经发生微环境。实验以成年虹鳟鱼为实验对象，研究了成年损伤视神经和一些大脑整合中枢再生过程，从而为揭示中枢神经系统损伤后的神经再生过程提供了新模型。

orcid: 0000-0003-0388-3147 (Evgeniya V. Pushchina)

关键词: 神经再生, 增殖, 凋亡, 视神经, 大脑, 鳟鱼, 放射状胶质细胞, 神经龛, 神经球

Abstract:

Fishes have remarkable ability to effectively rebuild the structure of nerve cells and nerve fibers after central
nervous system injury. However, the underlying mechanism is poorly understood. In order to address this
issue, we investigated the proliferation and apoptosis of cells in contralateral and ipsilateral optic nerves,
after stab wound injury to the eye of an adult trout Oncorhynchus mykiss. Heterogenous population of
proliferating cells was investigated at 1 week after injury. TUNEL labeling gave a qualitative and quantitative
assessment of apoptosis in the cells of optic nerve of trout 2 days after injury. After optic nerve injury,
apoptotic response was investigated, and mass patterns of cell migration were found. The maximal concentration
of apoptotic bodies was detected in the areas of mass clumps of cells. It is probably indicative of
massive cell death in the area of high phagocytic activity of macrophages/microglia. At 1 week after optic
nerve injury, we observed nerve cell proliferation in the trout brain integration centers: the cerebellum and
the optic tectum. In the optic tectum, proliferating cell nuclear antigen (PCNA)-immunopositive radial
glia-like cells were identified. Proliferative activity of nerve cells was detected in the dorsal proliferative
(matrix) area of the cerebellum and in parenchymal cells of the molecular and granular layers whereas local
clusters of undifferentiated cells which formed neurogenic niches were observed in both the optic tectum
and cerebellum after optic nerve injury. In vitro analysis of brain cells of trout showed that suspension cells
compared with monolayer cells retain higher proliferative activity, as evidenced by PCNA immunolabeling.
Phase contrast observation showed mitosis in individual cells and the formation of neurospheres which
gradually increased during 1–4 days of culture. The present findings suggest that trout can be used as a
novel model for studying neuronal regeneration.

Key words: nerve regeneration, proliferation, apoptosis, optic nerve, brain, radial glia cells, neurogenic niches, neurospheres, neural regeneration