Gene expression studies on human primordial germ cells and preimplantation embryos.
Research on the regulation of gene expression in human germ cells and preimplantation embryos is restricted due to the scarcity of samples and the requirement for highly sensitive molecular techniques to investigate the few cells available. To overcome these difficulties, we have developed a reliable procedure capable of generating amplified cDNA preparations from single cells. Using this procedure, we prepared cDNA from primordial germ cells (PGCs) isolated from the gonads of fetuses at 10 weeks gestation and from preimplantation embryos at the 1-cell, 4-cell, 8-cell and blastocyst stages. Our cDNA preparations allow us to investigate the expression profile of an almost unlimited number of different genes in the same sample preparation. This is of great advantage for studies of a panel of genes in a particular family or functional group, or with related mechanisms of regulation, e.g., developmental genes, oncogenes, cell cycle-control genes and imprinted genes. We have used these cDNA preparations in conjunction with differential display to identify genes specifically expressed in PGCs and preimplantation embryos in a sex- and developmental stage-specific manner. Genes specifically expressed in PGCs, oocytes and embryos were further analysed for their expression in embryonal carcinoma cells and in their differentiated derivatives following treatment by retinoic acid. Our strategy will disclose genes essential for gametogenesis and embryonic development which may only be expressed at certain stages of their development. The germ cell- and embryo-specific cDNA molecules, cDNA libraries and microarrays are a valuable resource for other researchers in this field.
Goto
T
T
Molecular Embryology Unit, Institute of Child Health, London, UK. Tetsuya.Goto@med.monash.edu.au
Holding
C
C
Daniels
R
R
Salpekar
A
A
Monk
M
M
D001755:Blastocyst / Q000737:chemistry / Q000378:metabolism*; D018076:DNA, Complementary / Q000032:analysis / Q000302:isolation & purification; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000737:chemistry / Q000378:metabolism*; D005865:Gestational Age; D006801:Humans; D008297:Male; D011247:Pregnancy
D018076:DNA, Complementary