Wroblewski V J, Harshman L G, Hanzlik T N, Hammock B D
Department of Entomology, University of California, Davis 95616.
Arch Biochem Biophys. 1990 May 1;278(2):461-6. doi: 10.1016/0003-9861(90)90285-7.
The tissue distribution, developmental control, and induction of juvenile hormone esterase (JHE) mRNA was examined in Heliothis virescens using an 800-base pair fragment of a JHE cDNA clone. Northern hybridization analysis of poly(A)+RNA from fat body and integument of fifth stadium larvae indicated the presence of a single JHE mRNA species having an estimated length of 3 kilobases. On Day 2 of the fifth stadium (L5D2), basal JHE mRNA levels were 3-fold higher in the integument than the fat body, which correlated with the higher specific activity of the enzyme in the integument at this time. However, JHE mRNA levels in the fat body on Day 4 of the fifth stadium were 9-fold higher than on Day 2, while mRNA levels in the integument remained the same. This endogenous increase in JHE mRNA and activity in the fat body occurred at the time of peak hemolymph JHE activity. JHE mRNA was not detected in third stadium larvae which have very low levels of JHE activity. Treatment of L5D2 larvae with the juvenile hormone mimic epofenonane resulted in a 7- and 14-fold increase in the level of JHE mRNA in the integument and fat body, respectively. The mRNA induced in both tissues was of the same estimated length as the constitutively expressed message. The data indicate that the developmental regulation and induction of JHE can occur at the level of mRNA. There is evidence that the fat body secretes more JHE than does the integument and could be the major source of hemolymph JHE.
利用幼年激素酯酶(JHE)cDNA克隆的一个800碱基对片段,检测了棉铃虫体内JHE mRNA的组织分布、发育调控及诱导情况。对五龄幼虫脂肪体和体壁的聚腺苷酸加尾RNA(poly(A)+RNA)进行的Northern杂交分析表明,存在一种估计长度为3千碱基的单一JHE mRNA种类。在五龄第2天(L5D2),体壁中的基础JHE mRNA水平比脂肪体高3倍,这与此时体壁中该酶较高的比活性相关。然而,五龄第4天脂肪体中的JHE mRNA水平比第2天高9倍,而体壁中的mRNA水平保持不变。脂肪体中JHE mRNA和活性的这种内源性增加发生在血淋巴JHE活性峰值时。在JHE活性非常低的三龄幼虫中未检测到JHE mRNA。用幼年激素类似物埃波烯诺处理L5D2幼虫,导致体壁和脂肪体中JHE mRNA水平分别增加7倍和14倍。在这两种组织中诱导产生的mRNA与组成型表达的信息具有相同的估计长度。数据表明,JHE的发育调控和诱导可发生在mRNA水平。有证据表明,脂肪体分泌的JHE比体壁多,可能是血淋巴JHE的主要来源。