Effects of Lhx8 in hippocampal cholinergic neural regeneration

doi:10.3969/j.issn.0529-1356.2013.06.001

Abstract

Abstract:

Objective To investigate the effect of Lhx8 in hippocampal cholinergic neural regeneration of SD rat after fimbria-fornix transection. Methods According to the different treatments, 72 SD rats were randomly divided into four groups: the normal group, the transected group, the overexpressed group and the transected combined with overexpressed group. On the 28th day after treatments, the sections of rat hippocampus of each group were prepared and stained with immunohistochemical methods to detect the number of newborn Lhx8 and ChAT positive cells. PCR and Western blotting were performed after total RNA and protein of hippocampus were obtained respectively. Results Compared with the normal group, the levels of Lhx8 mRNA and protein were significantly increased in other three groups and the differences were significant among the groups. No significant difference of the levels of ChAT mRNA was observed between the normal group and the transected group while it was significantly upregulated in the overexpressed and the transected combined with overexpressed groups. ChAT protein was detected in the transected, overexpressed and the transected combined with overexpressed groups and the differences were significant among the groups. There were newborn Lhx8 positive cholinergic neurons in hilus and subgranular zone of hippocampal dentate gyrus in the transected, overexpressed and the transected combined with overexpressed groups, with the most in the transected combined with overexpressed group, and the differences were significant among the groups. Conclusion Upregulation of Lhx8 can promote differentiation of neural stem cells into cholinergic neurons in hippocampus significantly, and Lhx8 may be related to cholinergic neural regeneration of hippocampus.

Key words: Lhx8, Cholinergic neuron, Neural regeneration, Hippocampus, Western blotting, Rat

SHI Jin-hong JIN Guo-hua* LI Hao-ming YI Xin QIN Jian-bing TIAN Mei-ling ZHANG Xin-hua. Effects of Lhx8 in hippocampal cholinergic neural regeneration[J]. AAS, 2013, 44(6 ): 729-734.

References

［1］Zhao Y, Marin O, Hermesz E, et al. The LIM-homeobox gene Lhx8 is required for the development of many cholinergic neurons in the mouse forebrain［J］. Proc Natl Acad Sci USA, 2003,100(15):9005-9010.
［2］Manabe T, Tatsumi K, Inoue M, et al. L3/Lhx8 is a pivotal factor for cholinergic differentiation of murine embryonic stem cells［J］. Cell Death Differ, 2007,14(6):1080-1085.
［3］Manabe T, Tatsumi K, Inoue M, et al. Knockdown of the L3/Lhx8 gene suppresses cholinergic differentiation of murine embryonic stem cell-derived spheres［J］. Int J Dev Neurosci, 2008,26(2):249-252.
［4] Zhang X, Jin G, Tian M, et al. The denervated hippocampus provides proper microenvironment for the survival and differentiation of neural progenitors［J］. Neurosci Lett, 2007,414(2):115-120.
［5］Zou LQ, Jin GH, Dong ChM, et al. Proliferation and diferentiation of the endogenous neural stem cells in hippocampus into neurons after fimbria fornix transection［J］. Su Zhou University Journal of Medical Science, 2007, 27(5):681-683.(in Chinese)
邹琳清, 金国华, 董传明, 等. 穹隆海马伞切割后海马内NSCs的增殖和向神经元的分化［J］. 苏州大学学报(医学版), 2007,27(5):681-683. 
［6］Dong ChM, Jin GH, Qin JB, et al. The effects of NGF on the proliferation and differentiation of endogenous NSCs in the hippocampus into neurons after fimbria fornix transaction ［J］. Acta Anatmica Sinica, 2007,38(6):642-646. (in Chinese)
董传明, 金国华, 秦建兵, 等. 神经生长因子对穹隆海马伞切割后海马自体神经干细胞增殖和向神经元分化的影响［J］. 解剖学报, 2007,38(6):642-646. 
［7］Jiang YY, Jin GH, Qin JB, et al. Change of Lhx8 mRNA expression in rat hippocampus after fimbria/fornix transection［J］. Chinese Journal of Anatomy,2008,31(2): 145-147.(in Chinese)
蒋媛媛, 金国华, 秦建兵, 等. 切割穹隆海马伞大鼠海马内Lhx8mRNA表达的变化［J］. 解剖学杂志, 2008, 31(2):145-147.
［8］Paxinos G, Watson C. The Rat Brain in Stereotaxic Coordinates ［M］. 2nd ed. Sydney: Academic Press, 1986: 21-24.
［9］Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR ［J］. Nucleic Acids Res, 2001, 29(9): e45.
［10］Frielingsdorf H, Thal LJ, Pizzo DP. The septohippocampal cholinergic system and spatial working memory in the Morris water maze［J］. Behav Brain Res, 2006, 168(1):37-46.［11］Mattson MP. Pathways towards and away from Alzheimer’s disease［J］. Nature, 2004, 430(7000): 631-639.
［12］Simmons DK, Pang K, Martindale MQ. Lim homeobox genes in the Ctenophore Mnemiopsis leidyi: the evolution of neural cell type specification［J］. Evodevo, 2012,3(1):2.
［13］Stivastava M, Larroux C, Lu DR, et al. Earlyevolution of the LIM homeobox gene family［J］. BMC Biol, 2010,8:4.
［14］Hung TM, Hu RH, Ho CM, et al. Downregulation of alpha-feto-protein expression by LHX4: a critical role in hepatocarcinogenesis［J］. Carcinogenesis,2011, 32(12):1815-1823.
［15］Colvin SC, Mullen RD, Pfaeffle RW, et al. LHX3 and LHX4 transcription factors in pituitary development and disease［J］. Pediatr Endocrinol Rev, 2009,(6 Suppl 2):283-290.
［16］Sheng HZ, Bertuzzi S, Chiang C, et al. Expression of murine Lhx5 suggests a role in specifying the forebrain［J］. Dev Dyn, 1997, 208(2):266-277.
［17］Shibaguchi T, Kato J, Abe M, et al. Expression and role of Lhx8 in murine tooth development［J］. Arch Histol Cytol, 2003, 66(1):95-108.
［18］Zhang Y, Mori T, Takaki H, et al. Comparison of the expression patterns of two LIM-homeodomain genes, Lhx6 and L3/Lhx8, in the developing palate［J］. Orthod Craniofac Res, 2002, 5(2):65-70.
［19］Pangas SA, Choi Y, Ballow DJ, et al. Oogenesis requires germ cell-specific transcriptional regulators Sohlh1 and Lhx8［J］. Proc Natl Acad Sci USA, 2006,103(21):8090-8095.
［20］Mori T, Yuxin Z, Takaki H, et al. The LIM homeobox gene, L3/Lhx8, is necessary for proper devdelopment of basal forebrain cholinergic neurons［J］. Eur J Neurosci, 2004,19(12): 3129-3141.
［21］Manabe T, Tatsumi K, Inoue M, et al. L3/Lhx8 is a pivotal factor for cholinergic differentiation of murine embryonic stem cells ［J］. Cell Death Differ, 2007, 14(6): 1080-1085.
［22］Manabe T, Tatsumi k, Inoue M, et al. Knockdown of the L3/Lhx8 gene suppresses cholinergic differentiation of murine embrgonic stem cell-derived spheres［J］. Int J Dev Neurosci, 2008, 26(2):249-252.

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