Establishment of a Purified Hippocampal Neuron Model in Naked Mole-rat and Preliminary Study on Its Hypoxic Tolerance

doi:10.3969/j.issn.1674-5817.2019.06.008

Abstract

Abstract: Objective To establish a method for purifying and culturing hippocampal neurons of naked mole-rats and to preliminarily explore their hypoxic tolerance characteristics. Methods Cells in hippocampus of naked mole-rats during 60-65 days of gestation were collected, and purified by the medium containing Neurobasal+2%B27+10 μmol/L 5-fluorouracil. Hippocampal neurons were maintained by neurobasal medium containing 2% B27. Cell growth was observed by inverted microscope after 6-7 days of adherent culture in vitro. Neuron-specific antibody was used to identify neuron-specific antibody. CCK8 kits were used to study on hypoxic tolerance of hippocampal neurons. Axonal length of naked mole-rat hippocampal neurons were measured by Image-Pro Plus 6.0. QPCR analysis was used to detect the hypoxia-inducible factor 1α (HIF1α) mRNA levels of hippocampal neurons under normoxia and hypoxia conditions. Results The hippocampal neurons obtained by this method exibits typical morphological characteristics of neurons, and immunocytochemical staining shows that the purified cells are NeuN-positive. The cell survival rate under hypoxia is more than twice of that under conditions with normal oxygen, and the axon lengths of cells in hypoxia conditions were smaller than those under nomoxia conditions. HIF1α levels of naked mole-rat hippocampal neurons were higher than those under normoxia conditions. There are statistically differences among these results (P<0.05). Conclusion The method established in this study is a simple and practical method for isolating and culturing hippocampal neurons of naked mole-rats in vitro, and has strong hypoxic tolerance.

Key words: Naked mole-rats, Hippocampus, Neurons, Hypoxia

CLC Number:

Q95-33

YANG Wen-jing, LI Yu, FENG Yan, LI Lei-chen, XU Yue-rong, LIU Pan, LIN Li-fang, SUN Wei, CUI Shu-fang. Establishment of a Purified Hippocampal Neuron Model in Naked Mole-rat and Preliminary Study on Its Hypoxic Tolerance[J]. Laboratory Animal and Comparative Medicine, 2019, 39(6): 467-472.

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