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LM23 is a novel member of the Speedy/Ringo family at the :rossroads of life and death of spermatogenic cell 被引量:4
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作者 Yi-Ming Cheng Mei-Ling Liu Meng-Chun Jia 《Asian Journal of Andrology》 SCIE CAS CSCD 2011年第3期446-452,513,共8页
LM23 is a gene specifically expressed in the testis of Rattus norvegicus, as previously reported by our laboratory. The aim of the study is to further investigate the biological function of LM23. Several bioinformatic... LM23 is a gene specifically expressed in the testis of Rattus norvegicus, as previously reported by our laboratory. The aim of the study is to further investigate the biological function of LM23. Several bioinformatic tools were utilized, including PROSITE and BLAST. To determine the subcellullar localization of LM23, a polyclonal antibody specific for LM23 was generated via the immunization of rabbits. The LM23gene was cloned from rat testis tissue, and LM23 protein was expressed in Escherichia co/i. The biological function of LM23 was analyzed with microarray analysis and immunohistochemistry, using a rat model of LM23 gene knockdown. The results suggested that LM23 belongs to the Speedy/Ringo family. LM23 regulated the GI/S and G2/M transitions of the cell cycle during spermatogenesis. Downregulation of the LM23gene during spermatogenesis could lead to the activation of both the Fas-FasL pathway and the mitochondrial pathway. These novel findings indicate that LM23 has a diverse array of functions that are important in both the life and death of the spermatogenic cell. 展开更多
关键词 cell cycle lm23 SPERMATOGENESIS spermatogenic cell
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LM23基因降调激活Fas-FasL和线粒体通路使精母细胞凋亡 被引量:3
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作者 成毅明 刘美玲 +1 位作者 王介东 贾孟春 《生殖医学杂志》 CAS 2011年第3期206-210,共5页
目的通过LM23基因RNA干扰(RNAi)动物模型,探讨LM23基因降调对大鼠生精细胞凋亡的影响。方法使用本实验室建立的LM23基因RNAi大鼠动物模型,分别通过苏木精-伊红染色(HE)和原位末端标记方法(TUNEL)检测LM23基因降调后生精细胞的凋亡发生情... 目的通过LM23基因RNA干扰(RNAi)动物模型,探讨LM23基因降调对大鼠生精细胞凋亡的影响。方法使用本实验室建立的LM23基因RNAi大鼠动物模型,分别通过苏木精-伊红染色(HE)和原位末端标记方法(TUNEL)检测LM23基因降调后生精细胞的凋亡发生情况;基因芯片技术,分析LM23基因降调后凋亡相关基因的差异表达;免疫组织化学技术检测LM23基因降调后caspase3蛋白分子在生精细胞中的表达。结果 TUNEL结果显示,LM23基因降调后大量精母细胞发生凋亡;基因芯片分析结果,LM23基因降调后Fas-FasL信号转导通路和线粒体信号转导通路被激活;免疫组织化学技术发现,LM23基因降调后,caspase3蛋白分子在大鼠睾丸精母细胞中的表达量显著增加。结论 LM23基因降调后,可能激活Fas-FasL信号转导通路和线粒体信号转导通路,从而导致大量精母细胞发生凋亡。 展开更多
关键词 lm23 生精细胞 凋亡 大鼠
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兔抗大鼠LM23蛋白合成肽多克隆抗体的制备与初步应用 被引量:2
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作者 成毅明 刘美玲 +2 位作者 石心泉 王介东 贾孟春 《生殖医学杂志》 CAS 2009年第3期279-283,共5页
目的制备大鼠精子发生相关蛋白LM23的合成肽多克隆抗体,对其特性进行初步鉴定,并对LM23多克隆抗体进行初步应用。方法利用生物信息学方法分析选择LM23蛋白的多肽序列。应用Fmoc法化学合成大鼠LM23蛋白N端第1~20和第274~291个氨基酸多... 目的制备大鼠精子发生相关蛋白LM23的合成肽多克隆抗体,对其特性进行初步鉴定,并对LM23多克隆抗体进行初步应用。方法利用生物信息学方法分析选择LM23蛋白的多肽序列。应用Fmoc法化学合成大鼠LM23蛋白N端第1~20和第274~291个氨基酸多肽(接近C端),经C18的RP-HPLC纯化后,通过高碘酸钠法,将纯化的LM23蛋白的多肽与KLH交联,免疫新西兰大耳白兔,获得LM23蛋白的多克隆抗体。用ELISA方法鉴定多克隆抗体效价。通过Western blot方法鉴定LM23多克隆抗体的特异性并研究LM23在大鼠睾丸中的表达;通过免疫组化方法研究LM23在大鼠睾丸组织和细胞中的定位。结果化学Fmoc法分别合成大鼠LM23蛋白N端第1~20,第274~291个氨基酸多肽,纯化后两条多肽纯度都达到90%以上,符合免疫用抗原标准。将多肽与KLH交联,用于免疫动物。经ELISA鉴定,两种多克隆抗体的效价分别达到1:64 000和1:128 000。应用大鼠睾丸组织切片进行LM23-18肽多克隆抗体的免疫组织化学研究,发现精母细胞有阳性颗粒反应,且LM23主要表达于细胞核。Western blot研究证实,LM23-18肽多克隆抗体可特异识别大鼠睾丸组织中相对分子量(Mr)约为36 kD的LM23蛋白。结论所制备的LM23合成肽多克隆抗体可用于ELISA、Western blot及免疫组化研究。LM23合成肽多克隆抗体的制备为进一步研究LM23的功能和作用机制提供了有力支持。 展开更多
关键词 lm23 合成肽 精子发生 多克隆抗体
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Role of LM23 in cell proliferation and apoptosis and its expression during the testis development 被引量:1
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作者 Qing Liu Ya-Juan Song +8 位作者 Li-Jun Meng Fen Hu Li-Xia Gou Chang-Hong Jia Hong-Mei Tang Wei Jie Wang Mi Li Xiu-Jun Zhang Meng-Chun Jia 《Asian Journal of Andrology》 SCIE CAS CSCD 2013年第4期539-544,I0010,共7页
LM23, a gene expressed specifically in the testis in a stage-specific manner, has a diverse range of functions that are important in both the life and death of spermatogenic cells. The aim of this study was to further... LM23, a gene expressed specifically in the testis in a stage-specific manner, has a diverse range of functions that are important in both the life and death of spermatogenic cells. The aim of this study was to further investigate the expression of LM23 in the developing rat testis and the biological function of LM23 in proliferation and antiapoptosis in vitro. Semiquantitative reverse transcription (RT)-PCR and real-time PCR were used to examine the expression of LM23 in testis at different developmental stages. The results suggested that LM23 mRNA levels in the testis increased progressively after birth. The role of LM23 in proliferation was analyzed with cell counting kit-8 (CCK8), colony-forming efficiency (CFE) and flow cytometry assays. The results indicated that ectopic expression of LM23 in 293T cells significantly promoted cell proliferation by increasing cell numbers in S phase. Several methods were used, including CCK8, annexin V and propidium iodide staining and western blotting, to determine the role of LM23 in apoptosis. The results showed that LM23 played a protective role in H202-induced apoptosis of 293T cells, mediated at least in part through the Akt/PI3K signal pathway. Taken together, these results provide new insights into the role of LM23 in the development of the testes andspermatogenesis. 展开更多
关键词 APOPTOSIS lm23 PROLIFERATION rat testis
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LM23 may regulate the G1/S and G2/M transitions of the cell cycle in rat spermatogenesis
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作者 成毅明 刘美玲 +2 位作者 詹皓 成玉 贾孟春 《生殖医学杂志》 CAS 2011年第B12期51-56,共6页
Objective:LM23(AF492385) is a gene specifically expressed in the testis of Rattus norvegicus previously reported by our laboratory.The aim of the study is to further investigate its biological function. Methods:Bioi... Objective:LM23(AF492385) is a gene specifically expressed in the testis of Rattus norvegicus previously reported by our laboratory.The aim of the study is to further investigate its biological function. Methods:Bioinformatic tools were utilized,including Protfun server and CPHmodles.The biological functions of LM23 were analyzed with microarray analysis,using a rat model of LM23 gene knock-down. Results:Protfun server shows that LM23 is likely a growth factor or Lyase.LM23 is more likely involved in regulatory functions of translation.The expressions of some genes related to the cell cycle were significantly changed after LM23 knock-down,as shown by microarray analysis. Conclusions:LM23 may regulate the G1/S and G2/M transitions of the cell cycle during spermatogenesis. 展开更多
关键词 精子发生过程 细胞周期 大鼠模型 G2 S期 生物学功能 基因敲除 基因芯片
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LM23基因沉默引起大鼠睾丸生精细胞凋亡 被引量:3
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作者 石心泉 贾孟春 刘美玲 《生殖医学杂志》 CAS 2010年第4期337-341,共5页
目的研究LM23基因敲低后生精细胞的凋亡状况及与凋亡相关基因表达改变。方法用TUNEL方法检测睾丸细胞的凋亡情况,用大鼠全基因组表达谱芯片检测LM23基因敲低后凋亡相关基因的表达改变。结果 TUNEL结果显示,LM23基因敲低侧大鼠睾丸生精... 目的研究LM23基因敲低后生精细胞的凋亡状况及与凋亡相关基因表达改变。方法用TUNEL方法检测睾丸细胞的凋亡情况,用大鼠全基因组表达谱芯片检测LM23基因敲低后凋亡相关基因的表达改变。结果 TUNEL结果显示,LM23基因敲低侧大鼠睾丸生精小管内大量生精细胞发生凋亡。大鼠全基因组表达谱芯片分析结果显示,许多促凋亡基因如Bcl-2家族的促凋亡基因表达上调,而抗凋亡基因如Faf1和Zfp91基因的表达明显下调。结论敲低大鼠的LM23基因,启动了Bcl-2家族介导的线粒体凋亡途径,引发了生精细胞发生大量凋亡。 展开更多
关键词 lm23基因 生精细胞 细胞凋亡 CASPASE-3
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