Changes of RUNX1T1 mRNA and protein expression in rat hippocampus after fimbria/fornix transection

doi:10.3969/j.issn.0529-1356.2013.06.007

Abstract

Abstract:

Objective To observe the temporal and spatial changes of RUNX1T1 mRNA and protein expression in rat hippocampus after fimbria/fornix transection at different time points. Methods Rat models were prepared by cutting fimbria fornix, and divided into normal group, cutting 3days, 7days, 14days, 21days and 28days groups. 1. The total hippocampal tissue mRNA was extracted, and the expression of RUNX1T1 mRNA was detected by real-time PCR; 2. The total protein of hippocampus was extracted, and the expression of RUNX1T1 protein was detected by western blotting; 3. Frozen brain sections were detected by RUNX1T1 immunofluorescence and Hoechst labeled, to observe the RUNX1T1/Hoechst double labeled positive cells in hippocampal dentate gyrus. Results Real-time PCR analysis showed that the RUNX1T1 mRNA expression in the hippocampus was significantly upregulated on the 3rd day after transection. Western blotting analysis indicated that the expression of RUNX1T1 protein was detected in the hippocampus of rats at basal condation, and the levels of RUNX1T1 protein were markedly increased at days 3 and 7, and peaked at day 3 after transection. In contrast, no siginificant change in the RUNX1T1 protein expression at days 14, 21 and 28. Immunostaining showed that the nember of RUNX1T1-positive cells was siginificantly higher at day 3 compared with control and decreased slowly after day 7. Conclusion The transient upregulation of RUNX1T1 mRNA and protein expression in the early phase after fimbria/fornix transection, and location in the hippocampal dentate gyrus granule lower, which may be related to the hippocampal neural regeneration.

Key words: RUNX1T1, Hippocampus, Fimbria/fornix transection, Neural regeneration, Immunofluorescence, Rat

ZOU Lin-qing JIN Guo-hua* WANG Hai-qin LI Hao-ming ZHU Pei-pei QIN Jian-bing. Changes of RUNX1T1 mRNA and protein expression in rat hippocampus after fimbria/fornix transection[J]. AAS, 2013, 44(6 ): 767-771.

References

［1］Malatesta P, Appolloni I, Calzolari F. Radial glia and neural stem cells［J］. Cell Tissue Res, 2008,331(1):165-178.
［2］Pollard SM, Conti L. Investigating radial glia in vitro［J］.Prog Neurobiol, 2007,83(1):53-67. 
［3］Li H, Jin G, Qin J, et al. Identification of neonatal rat hippocampal radial glia cellsin vitro［J］. Neurosci Lett, 2011,490(3):209-214.
［4］Zhao H, Jin G, Tian M, et al. Extract of deafferented hippocampus promotes in vitro radial glial cell differentiation into neurons［J］.Neurosci Lett, 2011,498(1):93-98. 
［5］Li H, Jin G, Qin J, et al. Generation and identification of rat fetal cerebral radial glia-like cells in vitro［J］. In Vitro Cell Dev Biol Anim, 2011,47(7):431-437. 
［6］Li H, Jin G, Qin J, et al. Characterization and identification of Sox2+ radial glia cells derived from rat embryonic cerebral cortex［J］. Histochem Cell Biol, 2011,136(5):515-526. 
［7］Shi JH，Jin GH，Li HM, et al. Gene expression of rest and RUNX1T1 in activiated hippocampal radial glial cells［J］. Acta Anatomica Sinica,2012,43(1):13-18. (in Chinese)
施金洪, 金国华, 李浩明, 等. 海马放射状胶质细胞激活后Rest和RUNX1T1基因的表达变化［J］. 解剖学报, 2012,43(1):13-18. 
［8］Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR［J］. Nucleic Acids Res, 2001,29(9):e45.
［9］Davis JN, McGhee L, Meyers S. The ETO (MTG8) gene family［J］. Gene, 2003,303:1-10.
［10］Schlenk RF, D?hner K, D?hner H. Acute myeloid leukemia. Genetic diagnostics and molecular therapy［J］. Internist (Berl), 2013,54(2):171-178. 
［11］Lam K, Zhang DE. RUNX1 and RUNX1-ETO: roles in hematopoiesis and leukemogenesis［J］. Front Biosci(Landmork Ed), 2012,17:1120-1139.
［12］Theriault FM, Nuthall HN, Dong Z, et al. Role for Runx1 in the proliferation and neuronal differentiation of selected progenitor cells in the mammalian nervous system［J］. J Neurosci, 2005,25(8):2050-2061.
［13］Yoshikawa M, Senzaki K, Yokomizo T, et al. Runx1 selectively regulates cell fate specification and axonal projections of dorsal root ganglion neurons［J］. Dev Biol, 2007,303(2):663-674.
［14］Koyano-Nakagawa N, Kintner C. The expression and function of MTG/ETO family proteins during neurogenesis［J］. Dev Biol, 2005,278(1):22-34.
［15］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.
［16］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］.Suzhou 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. 
［17］Shi J, Jin G, Zhu H, et al. The role of Brn-4 in the regulation of neural stem cell differentiation into neurons［J］. Neurosci Res, 2010,67(1): 8-17.
［18］Zou L, Jin G, Zhang X, et al. Proliferation, migration and neuronal differentiation of the endogenous NSCs in hippocampus after fimbria fornix transection［J］. Int J Neuroscie, 2010, 120(3): 192-200.

[1]	YANG Meng-yi NIU Shi-bo ZHANG Jing DANG Jie MA Zhan-bing LU Hong HUO Zheng-hao. Association between index finger and ring finger length ratios and polymorphism of homeobox A11 gene locus among Ningxia college students [J]. Acta Anatomica Sinica, 2024, 55(1): 62-66.
[2]	XIE Hui-lan FANG Fang LIN Yi. Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells [J]. Acta Anatomica Sinica, 2024, 55(1): 67-72.
[3]	WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen. Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia [J]. Acta Anatomica Sinica, 2024, 55(1): 3-9.
[4]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[5]	SU Peng WANG Xing-yi LIANG Jing-yan XIONG Tian-qing. Optimization and identification of a low density and high purity method for primary hippocampal neuron culture from fetal rats [J]. Acta Anatomica Sinica, 2024, 55(1): 113-119.
[6]	HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe . Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method [J]. Acta Anatomica Sinica, 2024, 55(1): 120-124.
[7]	XU Ya-ping YU Yue-xin CHEN Jin-fu WANG Ke-ke GUO Zhikun . Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats [J]. Acta Anatomica Sinica, 2023, 54(6): 716-721.
[8]	LIU Li-ping ZHU Meng-ni XU Hui . Effect of interleukin-6 on nucleated erythrocytes in lipopolysaccharide induced preeclampsia rats [J]. Acta Anatomica Sinica, 2023, 54(6): 722-729.
[9]	ZHAO Wei-ming LI Xiao YU Guo-ying WANG Gai-ping CHANG Cui-fang . Effect of serine protease inhibitor Kazaltype 1 on the proliferation of hepatocellular carcinoma cell and its mechanism [J]. Acta Anatomica Sinica, 2023, 54(6): 695-702.
[10]	WANG Wen-juan DING Yu-tong SU Hao REN Jie SUN Yan-rong WANG Han-fei LU Jia-li ZHANG Lin-qian BAI Yu QIN Li-hua. Changes of Nogo-A expression in hippocampus and striatum of rats under low estrogen condition [J]. Acta Anatomica Sinica, 2023, 54(6): 620-627.
[11]	LIANG Pan-pan YU Ai-mei DU Jing KOU Wen-hui WANG Huan-huan SONG Ai-xia. Inhibitory effect of sodium ferulate on inflammatory response in migraine rats based on c-Jun N-terminal kinase/p38 mitogen-activated protein kinase signaling pathway [J]. Acta Anatomica Sinica, 2023, 54(6): 652-659.
[12]	YAO Su-hua YAN Jun XIE Fang LIN Chun-hua HUANG Zheng-chun. Nerve repair effect of olanzapine-mediated cyclic AMP response element binding protein/brain-derived neurotrophic factor/receptor tyrosine kinase receptors B pathway on schizophrenic rats [J]. Acta Anatomica Sinica, 2023, 54(6): 660-667.
[13]	XU Ying LIU Bin ZHENG Xing HE Zong-zhao. Role and mechanism of complement C3a receptor in the cognitive impairment of septic rats induced by cecal ligation and perforation [J]. Acta Anatomica Sinica, 2023, 54(6): 668-675.
[14]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.
[15]	LI Xia ZHU Shi-jie TANG Zhong-sheng LUO Ya-fei FAN Rui-juan XIE Gao-yu KOU Yun-fang LU Ying. Mechanism of electroacupuncture regulating the ligand pathway of tyrosine kinase receptor in improving vascular dementia [J]. Acta Anatomica Sinica, 2023, 54(6): 689-694.