Wang Duo, Li Mengmeng, Ma Jingyi, Wang Xuanxuan, Liu Jingze
Hebei Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology, Hebei Collaborative Innovation Center for Eco-Environment, Ministry of Education Key Laboratory of Molecular and Cellular Biology, College of Life Sciences, Hebei Normal University, Hebei province, 050024, Shijiazhuang, Hebei province, China.
Exp Appl Acarol. 2023 Jan;89(1):105-115. doi: 10.1007/s10493-022-00774-z. Epub 2023 Jan 19.
The effects of temperature on the expression patterns and enzyme activity of cathepsin B (HlCatB), cathepsin D (HlCatD) and acid phosphatase (HlACP) during the embryo development of Haemaphysalis longicornis (bisexual population) were investigated in this study. Eggs were exposed to 20 °C (low temperature), 26 °C (normal temperature), and 30 °C (high temperature) immediately after laying, and collected on odd days of embryo development to measure HlCatB, HlCatD and HlACP gene expression using quantitative real-time PCR, as well as three enzyme activities using spectrophotometry. Then the associations between mRNA expression levels of three enzymes and their enzyme activities were assessed. Compared with normal temperature, the mRNA expression peaks of HlCatB were higher and appeared later at low and high temperatures and the activity of HlCatB increased on most days of embryonic development at high temperature. As for HlCatD, the expression peak appeared later at low temperature, but earlier at high temperature. The activity peaks of HlCatD were lower and appeared earlier at low and high temperatures. As for HlACP, the expression peak was higher and appeared later at low temperature, whereas it formed no prominent peak at high temperature. The activity peak of HlACP was higher at low temperature, but lower at high temperature. The linear regression analysis showed that activities of three enzymes were associated with their mRNA expression levels (P < 0.05). Three enzymes are involved in the embryo adaptation to temperature stress. Moreover, the mRNA expression level may be another factor affecting its enzyme activity.
本研究调查了温度对长角血蜱（两性种群）胚胎发育过程中组织蛋白酶B（HlCatB）、组织蛋白酶D（HlCatD）和酸性磷酸酶（HlACP）表达模式及酶活性的影响。产卵后，将卵立即置于20℃（低温）、26℃（常温）和30℃（高温）环境中，并在胚胎发育的奇数天收集样本，采用定量实时PCR检测HlCatB、HlCatD和HlACP基因表达，同时采用分光光度法测定三种酶的活性。然后评估三种酶的mRNA表达水平与其酶活性之间的相关性。与常温相比，HlCatB的mRNA表达峰值在低温和高温下更高且出现时间更晚，并且在高温下胚胎发育的大多数天数里HlCatB的活性增加。至于HlCatD，其表达峰值在低温下出现时间更晚，但在高温下出现时间更早。HlCatD的活性峰值在低温和高温下更低且出现时间更早。至于HlACP，其表达峰值在低温下更高且出现时间更晚，而在高温下未形成明显峰值。HlACP的活性峰值在低温下更高，但在高温下更低。线性回归分析表明，三种酶的活性与其mRNA表达水平相关（P < 0.05）。三种酶参与胚胎对温度胁迫的适应。此外，mRNA表达水平可能是影响其酶活性的另一个因素。
