Abstract
Purpose
The present study was conducted to investigate the expression and possible role of the c-mycprotooncogene in preimplantation embryos by using reverse-transcriptase/polymerase chain reaction (RT-PCR) technique and microinjection of synthetic antisense c-mycoligonucleotide probe, respectively. Total RNA was extracted from oocytes and two cell-, four cell-, early morula-, late morula-, early blastocyst-, and late blastocyst-stage embryos, and cDNA was constructed using MMLV reverse transcriptase. Sense primer (P 1)and antisense primer (P 2)used were based on the c-mycgene sequence bp 1609–1629 and bp 3279–3299, respectively, that span a 1.37-kb intron. PCR-amplified products of cDNA from oocyte-, two cell-, four cell-, early and late morula-, and blastocyst-stage embryos demonstrated the expected 313-bp product in Southern blot hybridization using a c-mycspecific DNA probe, with an indication of lower levels in oocytes and early morulae.
Results
Cytoplasmic injection of the antisense c-mycoligonucleotide probe (P 2)and not the sense probe (P 1)into pronuclear-stage zygotes caused a significant (P=0.02 to 0.0001) inhibition of development to blastocysts in a concentration-dependent manner, with a maximal inhibition at the first cleavage of zygotes to two cell-stage embryos. There was no effect on the P 2 antisense injection on pronucleus formation.
Conclusion
These results indicate that the c-mycprotooncogene is expressed in preimplantation embryos and may have an essential role in normal embryogenesis in mice.
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Naz, R.K., Kumar, G. & Minhas, B.S. Expression and role of c-myc protooncogene in murine preimplantation embryonic development. J Assist Reprod Genet 11, 208–216 (1994). https://doi.org/10.1007/BF02211810
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DOI: https://doi.org/10.1007/BF02211810