Identification of a novel testis-specific gene and its potential roles in testis development／spermatogenesis

Aim: To identify and characterize a novel gene with potential roles in testis development and spermatogenesis.Methods: A cDNA microarray was constructed from a human testis large insert cDNA library and hybridized with probes of human or mouse adult and fetal testes. Differentially expressed genes were isolated and sequenced. RT-PCR was used to test the tissue distribution of the genes of interest and in situ hybridization was performed to localize the gene expression in the mouse testis. A range of bioinformatical programs including Gene Runner, SMART, NCBI Blast and Emboss CpGPlot were used to characterize the new gene‘s feature. Results: A novel testis-specific gene,NYD-SPS, was differentially expressed in fetal and adult testes. The deduced protein structure of NYD-SP5 was found to contain an IQ motif (a short calmodulin-binding motif containing conserved lie and Gin residues), a Carbamate kinase-like domain, a Zn-dependent exopeptidase domain and a lactate dehydrogenase (LDH) C-terminal-like domain. RT-PCR analysis revealed that NYD-SP5 was predominantly expressed in the testis but not in other 15 tissues examined. In situ hybridization and RT-PCR examinations revealed that the expression of NYD-SP5 was confined in the male germ cell but not present in the somatic cell in the testes. Conclusion: NYD-SP5 is a newly found testisspecific gene with potential roles in testis development and spermatogenesis through a calmodulin-activated enzyme.     

Screening the stage-specific expression gene from different germ cells of rat

<正> Objective:To screen the stage-specific expression genes from rat spermatogonia,pachytene spermatocytes and round spermatids.Methods:Highly purified spermatogonia were isolated from 9-day-old rats,pachytene spermatocytes and round spermatids from adult rats by sedimentation velocityat unit gravity,using 2%-4% BSA gradient in DMEM/F12 medium.A mRNA differen-tial display method was used for screening the stage-specific expression gene.Results:Nineteen differentially/ expressed cDNA fragments were obtained.Afterexcluding the false positive cDNA fragments by dot blot,13 cDNAs were selected toclone and sequence.To obtain longer cDNAs,six ESTs were used to screen the rattestis λ-zap Ⅱ cDNA library.Two longer cDNA fragments,designated as LY21 andLM66,were obtained.The analysis with DNAMAN software indicated that LY21 had along open reading frame coding 372 amino acids while LM66 had no long open readingframe.LY21 were highly homologous with hnRNP H1.To observe the expression pat-terns of LY21 gene in the testicular cells,we performed in situ hybridization on testissections from adult rats.The LY21 gene expression was found in the spermatogonia andprimary spermatocytes.Conclusion:This study indicated that LY21 gene was associated with spermatogen-esis.Further studies will be needed to explore the function of LY21.     

Cloning of differentially expressed genes following lipopolysaccharide stimulation in human umbilical vein endothelial cells

Objective: To clone the differentially expressed genes in human umbilical vein endothelial cells (HUVEC)stimulated by lipopolysaccharide (LPS). Methods : Two-directional ( forward and backward)suppression subtractive hybridization ( SSH ) was performed on HUVEC cultured in either standard media or treated for 6 hours with LPS (100 ng/ml). To restrict the number of false-positive clones, colony dot hybridization was used to further verify the differentially expressed cDNA clones. Positive clones were sequenced. Results: These analyses have identified both novel and known genes whose expression is influenced by LPS.The known genes include a group related to proinflammatory events, a group related to cellular apoptosis and proliferation, a group related to protein synthesis and cytoskeletal rearrangment, and a group related to energy metabolism and signal transduction. Conclusions: SSH is a powerful technique of high sensitivity for the detection of differential gene expression in HUVEC stimulated by LPS.     

Metastases-related genes in the classification of liver and peritoneal metastasis in human gastric cancer

INTRODUCTION: With the aim of identifying metastases-related genes in gastric cancer, we performed a broad analysis of differential gene expression between low-metastatic parental cell lines and established highly metastatic sublines. MATERIALS AND METHODS: We established novel cell lines, AZ-H5c, NUGC-3H5, and TMK-1H7, with a high potential of liver metastasis, and AZ-P7a, NUGC-3P4T, and TMK-1P4a, with a high potential of peritoneal metastasis. These cell lines were derived from low-metastatic parental AZ-521, NUGC-3, and TMK-1 cell lines, respectively. Furthermore, to investigate different levels of gene expression implicated in metastatic potentials in gastric cancer, we investigated approximately 2000 expressed genes in each cell line using a DNA microarray. RESULTS: Varieties of genes were up-regulated or down-regulated in highly metastatic liver and peritoneal cell lines. Fifty-eight genes, including the transferrin receptor, ras-related rho, and osteopontin, and 22 genes, including apolipoprotein E and inhibin A-submit, were up-regulated and down-regulated in two or three liver metastatic sublines. On the other hand, 19 genes, the transferrin receptor, c-fos, and RANTES, and 26 genes, including MAC25, PISSLRE, and RNA polymerase, were up-regulated and down-regulated in two or three peritoneal metastatic sublines. CONCLUSION: How gene expression is implicated in gastric cancer metastasis has never been thoroughly explained, and further studies are necessary to understand the involvement of genes in cancer metastasis more thoroughly. We hope that our highly metastatic liver and peritoneal experimental models are helpful for further study and gene therapy of human gastric cancer.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.     

特发性高草酸尿症大鼠肝脏差异表达基因的筛选研究

目的 从基因水平探索特发性高草酸尿症(IH)的发病机制,筛选可能导致其发病的相关基因.方法应用含有26 962条大鼠全基因组寡核苷酸芯片,研究3只IH模型大鼠和3只正常大鼠肝脏组织基因表达谱的差异性,同时进行生物信息学分析.结果 在IH模型大鼠与正常大鼠肝脏组织之间存在差异表达基因147条,其中上调基因123条,下调基因24条,包括细胞受体、免疫相关、细胞信号和传递蛋白、代谢蛋白、发育相关等多种基因.结论 基因芯片能有效筛选出IH模型大鼠肝脏中的差异基因,显著的基因差异表达与其发病机制可能存在相关性.