RESUMEN
AIM: To investigate the expression of Spindlin 1 (Spin 1) isoform2 and assess its function in mouse testis. METHODS: First, reverse-transcription polymerase chain reaction (RT-PCR) was used to determine whether Spin1 isoform2 is present in mouse testis. Then the expression patterns of the isoform between newborn and adult mice testes were compared by immunoblot analysis. Finally, the diversity of its localization in mice testes at different ages (days 0, 7, 14, 21, 28 and 60) was observed by immunohistochemistry. The localization of the protein in mouse sperm was also investigated by immunofluorescence. RESULTS: The RT-PCR results show that Spin1 isoform2 is present in mouse testis. As shown by immunoblot analysis, the isoform was more highly expressed in adult testes compared with newborn testes. Interestingly, Spin1 isoform2 did not show up in the cytoplasm of primary spermatocytes until day 14. Also, the protein exists at the tail of the mouse sperm. CONCLUSION: Spin1 isoform2 is a protein expressed highly in adult testis, which might be involved in spermatogenesis and could be necessary for normal sperm motility.
Asunto(s)
Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Testículo/fisiología , Factores de Edad , Animales , Animales Recién Nacidos , Western Blotting , Citoplasma/metabolismo , Femenino , Inmunohistoquímica , Masculino , Meiosis/fisiología , Ratones , Ratones Endogámicos ICR , Embarazo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Motilidad Espermática/fisiología , Espermatogénesis/fisiologíaRESUMEN
Hemin-catalytic decomposition of artemisinin (qinghaosu, QHS) was studied using pyronine B (PB) as an indicator. The interaction between hemin and QHS was an enzyme-substrate model, and the action sites were the endoperoxide moiety of QHS and the central metal ion of enzyme respectively. The kinetic catalytic constant depends upon enzyme and substrate concentrations, and the Michaelis-Menten parameters Km, Vmax and Kcat was 8.4 x 10(-5) mol x L(-1), 7.4 x 10(-6) mol x L(-1) s(-1) and 50.23 s(-1) respectively. The catalytic activity of hemin was inhibited in the presence of deactivated agents and at high temperature. Under optimal conditions, the change in fluorescence intensity (Fo-F) of pyronine B was proportional to the QHS concentration from 0.0 to 1.27 x 10(-6) mol x L(-1), and the detection limit (3sigma) was as low as 2.3 x 10(-8) mol x L(-1). The proposed method was applied to detect the concentration of QHS in the media of plasma and urine.