Wiehart U I M, Nicolson S W, Eigenheer R A, Schooley D A
Department of Zoology, University of Cape Town, Rondebosch 7701, South Africa.
J Exp Biol. 2002 Feb;205(Pt 4):493-501. doi: 10.1242/jeb.205.4.493.
Fluid secretion by insect Malpighian tubules is controlled by haemolymph-borne factors. The mealworm Tenebrio molitor provides the first known example of antagonistic interactions between endogenous neuropeptides acting on Malpighian tubules. The two corticotropin-releasing-factor (CRF)-related diuretic peptides previously isolated from Tenebrio molitor, Tenmo-DH(37) and Tenmo-DH(47), were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC(50) values of 0.12 nmol l(-1) and 26 nmol l(-1) respectively. However, no synergistic or additive effect was observed when these two peptides were tested simultaneously. We then investigated antagonism between second messengers: dose-response curves were constructed for stimulation of Tenebrio molitor tubules by cyclic AMP and their inhibition by cyclic GMP. When both cyclic nucleotides were included in the bathing Ringer, the stimulatory effect of cyclic AMP was neutralised by cyclic GMP. Similarly, the stimulatory effect of Tenmo-DH(37) was reversed on addition of an antidiuretic peptide (Tenmo-ADF), which was recently isolated from Tenebrio molitor and acts via cyclic GMP. The cardioacceleratory peptide CAP(2b), originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC(50) value of 85 nmol l(-1). This inhibitory effect was reversed by Tenmo-DH(37). Endogenous diuretic and antidiuretic peptides, effective at low concentrations and acting via antagonistic second messengers, have the potential for fine control of secretion rates in the Malpighian tubules of Tenebrio molitor.
昆虫马氏管的液体分泌受血淋巴中携带的因子控制。黄粉虫是已知的内源性神经肽作用于马氏管时产生拮抗相互作用的首个例子。先前从黄粉虫中分离出的两种促肾上腺皮质激素释放因子(CRF)相关利尿肽,即黄粉虫利尿肽(Tenmo-DH(37))和黄粉虫利尿肽(Tenmo-DH(47)),被发现能在体外以剂量依赖的方式刺激黄粉虫马氏管,其半数有效浓度(EC(50))值分别为0.12 nmol l(-1)和26 nmol l(-1)。然而,当同时测试这两种肽时,未观察到协同或相加效应。然后我们研究了第二信使之间的拮抗作用:构建了环磷酸腺苷(cAMP)刺激黄粉虫马氏管及其被环磷酸鸟苷(cGMP)抑制的剂量反应曲线。当两种环核苷酸都包含在浴用林格液中时,cAMP的刺激作用被cGMP中和。同样,在添加一种抗利尿肽(黄粉虫抗利尿因子,Tenmo-ADF)后,黄粉虫利尿肽(Tenmo-DH(37))的刺激作用被逆转,该抗利尿肽最近从黄粉虫中分离出来,通过cGMP起作用。最初从烟草天蛾中分离出的促心加速肽CAP(2b)也能增加细胞内cGMP水平,并抑制黄粉虫马氏管的液体分泌,其半数有效浓度(EC(50))值为85 nmol l(-1)。这种抑制作用被黄粉虫利尿肽(Tenmo-DH(3(7)))逆转。内源性利尿和抗利尿肽在低浓度下起作用,并通过拮抗的第二信使发挥作用,具有精确控制黄粉虫马氏管分泌速率的潜力。