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Department of Anatomy and Reproductive Biology,4 John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii 96822
Center for Research on Reproduction and Women's Health,5 University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
Quantitative real-time polymerase chain reaction assay was used to compare the temporal transcriptional activation and mRNA removal for a number of genes in mouse embryos derived by round spermatid injection (ROSI) or intracytoplasmic sperm injection. A number of marker genes with widely different cellular functions were analyzed. Similar patterns of activation were found for the transcription factor Oct 4, the translation initiation factor eukaryotic initiation factor 1A, the L1 ribosomal protein, the chromatin modifying protein histone deacetylase 1, the enzyme hypoxanthine phosphoribosyl transferase, the murine endogenous retrovirus-like element, and the repetitive DNA LINE retrotransposons. Expression of the retrovirus-like mobile element intracisternal A particle, however, was markedly elevated from the two-cell to the blastocyst stages in ROSI embryos. Analyses performed for various paternal mRNAs introduced into the oocyte by the round spermatid, including protamines 1 and 2, transition protein 2, ropporin, and glyceraldehydes 3-phosphate dehydrogenase, revealed all were removed from the preimplantation embryos, albeit with distinct temporal patterns.
2 Correspondence: Norman B. Hecht, Center for Research on Reproduction and Women's Health, University of Pennsylvania Medical School, 1310 Biomedical Research Building II/III, 421 Curie Blvd., Philadelphia, PA 19104-6142. FAX: 215 573 5408; nhecht{at}mail.med.upenn.edu
3 These two authors contributed equally to this research
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