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Differentiation of HT22 neurons induces expression of NMDA receptor that mediates homocysteine cytotoxicity

NMDA受体 碘化丙啶 细胞生物学 细胞分化 程序性细胞死亡 化学 受体 谷氨酸受体 兴奋毒性 细胞凋亡 生物 生物化学 基因
作者
Zhongyan Zhao,Run Lu,Bei Zhang,Jun Shen,Liguo Yang,Songhua Xiao,Jun Liu,William Z. Suo
出处
期刊:Neurological Research [Informa]
卷期号:34 (1): 38-43 被引量:35
标识
DOI:10.1179/1743132811y.0000000057
摘要

Introduction: Neurotoxic homocysteine (Hcy) is thought to be an independent risk factor for neurodegenerative diseases, including Alzheimer's disease. This study is to determine whether HT22 cells, a murine hippocampal neuronal model, can be used as an in vitro model, besides the primary neuronal cultures, to investigate the effects of Hcy.Materials and methods: MTS assay and Hoechst 33342/propidium iodide discrimination were used to assess the cell viability and cell death on undifferentiated and differentiated HT22 cells. Semi-quantitative reverse transcription polymerase chain reaction and western blot were used to determine the expression of N-methyl D-aspartate (NMDA) receptor.Results: We found that undifferentiated and differentiated HT22 cells responded to Hcy toxicity differentially, with the undifferentiated cells resistant while the differentiated cells sensitive. The underlying mechanism appeared to be the differential expression levels of NMDA glutamate receptor between the undifferentiated and differentiated cells. Similar to what have been observed in primary neuronal cultures, the Hcy toxicity in the differentiated HT22 cells was largely attenuated by NMDA receptor antagonists, MK-801 and memantine.Conclusion: These results suggest for the first time that the differentiation of HT22 cells could induce the expression of NMDA receptors, which lead to Hcy mediate concentration-dependent apoptosis-necrotic continuum of HT22 cell death. The differentiation status of the HT22 cells is important for modeling neurons in vitro, with the differentiated HT22 neurons resembling more characteristics of primary hippocampal neurons while the undifferentiated HT22 cells being proliferating neuronal precursor cells. The differentiated HT22 neurons can be used as a platform to study Hcy toxicity.
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