Identification of Differentially Expressed Genes of Primary
Cell Research (2004); 14(6):507-512 ARTICLE http://www.cell-research.com Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique Gang LIANG1,4, Xiao Dong ZHANG1, Lu Jing WANG1, Yu Shen SHA2, Jian Chao ZHANG2, Shi Ying MIAO1, Shu Dong ZONG2, Lin Fang WANG1,*, S.S. KOIDE3 1National Laboratory Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, 5 Dong Dan San Tiao, 100005 Beijing, China 2National Research Institute for Family Planning, WHO Collaboration Center for Research in Human Reproduction, Beijing, 12 Da Hui Si, 100081 Beijing, China 3Center for Biomedical Research, Population Council, 1230 York Avenue, New York, NY 10021, USA 4Chinese National Human Genome Center, Beijing, 3-707 North Yong Chang Road BDA, Beijing 100176, China ABSTRACT The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differen- tiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals.
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