In total, 4165 medaka (Oryzias latipes) oocytes were injected with three DNA constructs separately, and results showed that exogenous lacZ expression was transient and stage-dependent. The initiation of the transgene expression was at the mid-blastula stage for embryos derived from oocytes injected with pmiwZ, containing the long terminal repeat (LTR) of the Rous sarcoma virus, and with pCAGGS-lacZ, containing the enhancer and promoter of the immediate early gene of the human cytomegalovirus, respectively, whereas embryos derived from oocytes injected with pMoZtk, containing the LTR of the Moloney murine leukemia virus, started expression at the late-blastula stage. These reveal that the earliest onset of the exogenous lacZ gene should be by the mid-blastula stage. Therefore the mid-blastula transition phenomenon in embryogenesis known in other animal species exists in medaka embryos.