RESUMO
BACKGROUND: The tick Haemaphysalis longicornis exhibits two separate reproductive populations: bisexual and parthenogenetic, which have diploid and triploid karyotypes, respectively. The parthenogenetic population can undergo engorgement without copulation and produce viable female-only offspring with a longer incubation period than the bisexual population. Three enzymes, cathepsin B, cathepsin D and acid phosphatase, were found to be involved in vitellin degradation during the embryonic development of bisexual H. longicornis. However, the expression and activity profiles of these enzymes during the embryonic development of parthenogenetic ticks remain unknown. In the present study, the transcriptional expression profile, enzyme activity and roles in embryogenesis of the three enzymes during the embryonic development of parthenogenetic H. longicornis were investigated. METHODS: Quantitative real-time polymerase chain reaction (qPCR) and fluorescence detection were used to analyze the dynamic changes in the three enzymes during embryogenesis. The roles of the three enzymes during embryogenesis were also explored using RNA interference (RNAi). RESULTS: The three enzymes were all expressed during embryonic development in parthenogenetic H. longicornis. The expression of cathepsin B was highest on day 15, whereas that of cathepsin D was highest on day 3 and the peak of acid phosphatase expression occurred on day 9. The activity of cathepsin B was highest on day 3 and lowest on day 5, then gradually increased and remained stable. Cathepsin D activity was highest on day 1 and showed a gradually decreasing trend, whereas acid phosphatase showed the opposite trend and reached a peak on day 23. RNA interference experiments in engorged female ticks revealed that there was no significant difference in the number of eggs laid, but the hatching rate of the eggs was significantly decreased. CONCLUSION: The three enzymes all play important roles in embryonic development of H. longicornis, but the expression patterns and changes in the activity of the enzymes in the bisexual and parthenogenetic populations are different. The results will help a better understanding of the similarities and differences underlying embryonic development in the bisexual and parthenogenetic populations and contribute to the future exploration of the development of the parthenogenetic population of H. longicornis.
Assuntos
Fosfatase Ácida/metabolismo , Vetores Aracnídeos/embriologia , Catepsina B/metabolismo , Catepsina D/metabolismo , Ixodidae/embriologia , Partenogênese/fisiologia , Fosfatase Ácida/genética , Animais , Vetores Aracnídeos/enzimologia , Vetores Aracnídeos/fisiologia , Catepsina B/genética , Catepsina D/genética , Clonagem Molecular , Desenvolvimento Embrionário , Feminino , Inativação Gênica , Ixodidae/enzimologia , Ixodidae/fisiologia , Oviposição/fisiologia , Interferência de RNA/fisiologia , RNA de Cadeia Dupla/fisiologia , RNA Mensageiro/metabolismo , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Tempo , Vitelinas/metabolismoRESUMO
BACKGROUND: Three main enzymes including cathepsin B, cathepsin D and acid phosphatase are involved in vitellin degradation, which is a major biochemical event of the embryonic development and can provide nutrients and metabolites for tick embryos. In the present study, the mRNA expression profiles and enzymatic activity of cathepsin B, cathepsin D and acid phosphatase were investigated during embryonic development in the tick Haemaphysalis longicornis. RESULTS: The results revealed that all three enzymes were expressed throughout embryonic development. Both cathepsin B and acid phosphatase transcripts were accumulated during the first four days. Cathepsin B reached its highest expression on day 5, whereas the peak expression of acid phosphatase and cathepsin D occurred on day 11. The highest activity of cathepsin B was observed on the first day of egg development, whereas cathepsin D reached its highest activity on day 13. Acid phosphatase activity increased gradually during the first five days and then remained stable until the end of egg development. CONCLUSIONS: Three enzymes were expressed and activated in eggs, and also presented different dynamic changes with the development of embryos. The profiles of both mRNA expression and enzymatic activity of these enzymes indicate that they are controlled orderly and play multiple roles during embryonic development in ticks.
Assuntos
Fosfatase Ácida/genética , Catepsina B/genética , Catepsina D/genética , Regulação da Expressão Gênica no Desenvolvimento , Ixodidae/enzimologia , Fosfatase Ácida/metabolismo , Animais , Catepsina B/metabolismo , Catepsina D/metabolismo , Feminino , Ixodidae/embriologia , Ixodidae/genéticaRESUMO
Coxiella-like endosymbiont (CLS-Hl) is a primary endosymbiont of Haemaphysalis longicornis. CLS-Hl infects tick special tissues and its prevalence is 100% in ovaries and Malpighian tubules. Tetracycline was injected into females, which then fed on rabbits also treated with tetracycline. The densities of CLS-Hl were measured by semi-quantitative PCR. CLS-Hl densities in ovaries and Malpighian tubes of H. longicornis had significant effects on engorged weight, feeding time, number of eggs, oviposition period, and hatching period. These findings suggested that CLS-Hl plays a role in the reproduction and development of H. longicornis.
