Isolation and identification of a diuretic hormone from Zootermopsis nevadensis☆
Introduction
The control of water balance is of fundamental importance in maintenance of homeostasis in insects [4], [20]. A variety of factors have now been chemically identified which stimulate basal secretion of ions, and hence urine, by the Malpighian tubules [7], [9], [23]. The two best studied families of factors stimulating diuresis are the corticotropin-releasing factor (CRF)-related DH and the myokinins. The former are similar to a family of peptides including sauvagine, CRF, urotensin I, and urocortin, and have been identified in nine species from five insect orders [3], [5], [6], [11], [12], [13], [14], [15], [16], [17], [18]. Their action on the Malpighian tubules (Mt) is mediated via cyclic AMP (cAMP) as second messenger [8].
M. sexta was the first species found to possess two CRF-related DH, Manse-DH [14] and Manse-DPII [3]. Manse-DPII has eleven less amino acids than Manse-DH and shows lower similarity to other CRF-related DH than does Manse-DH. Subsequently we identified two CRF-related DH from the coleopteran Tenebrio molitor, Tenmo-DH37 [13] and Tenmo-DH47 [12], again with the shorter peptide showing lower similarity to other CRF-related DH than does Tenmo-DH47 (Fig. 3). Very recently we reported the structures of two CRF-related DH from the sphingid moth, Hyles lineata: Hylli-DH41 and Hylli-DH30 [10]. It is possible that many insects may have two CRF-related DH; the biological significance of having two DH that use the same second messenger is not known.
We also reported recently the existence of two DH in the Pacific beetle cockroach, Diploptera punctata [11]. One of these two DH, Dippu-DH31, was found to be ineffective in elevating levels of cAMP produced by Malpighian tubules of M. sexta; previously the only DH found to be incapable of stimulating Malpighian tubules of this species were the two T. molitor DH, perhaps due to their C-terminal free carboxylate group [12], [13]. The 31 residue Dippu-DH31 is more similar to calcitonin than to the CRF superfamily, and elevates cAMP production by tubules of Schistocerca americana, Locusta migratoria, and D. punctata [11]. Its actions in D. punctata are more consistent with its causing an elevation of intracellular Ca2+ rather than cAMP [11].
We now report the characterization of two factors from the dampwood termite Z. nevadensis which have strong biological activity on Malpighian tubules of M. sexta. We have identified the more hydrophobic of these factors as a 46 residue neuropeptide which we designate Zoone-DH. We could not isolate the more hydrophilic, more basic of the two factors in two attempted isolation procedures. We presume that the factor which we could not isolate is another CRF-related DH, due to its high activity on M. sexta Malpighian tubules; to date no “short” CRF-like DH has been isolated from the hemimetabolous insects, which are more primitive than holometabolous insects from which the short CRF-like DH have been identified.
Section snippets
Experimental animals
Z. nevadensis were collected from April to September in the Toiyabe National Forest from Section 23, Township 20 North, Range 17 East, approximately 5.5 km north of Verdi, Nevada, and about 5 km west of the California-Nevada border. Soldier and reproductive insects were removed and the remaining animals were decapitated, the heads collected in liquid N2, and then stored at 27°C until extraction. M. sexta were reared at 25°C during the day, −80°C during the night, on a 16:8 light:dark cycle;
Results
We used Mt of adult male M. sexta to assay fractions, as the DH receptor of this organism has been shown to be quite non-specific in its sensitivity to members of the CRF-like DH [2], save for those with a carboxyl-terminal free acid residue, such as Tenmo-DH37 and -DH47. Four other CRF-like DH have been isolated using this heterologous bioassay system [6], [10], [11]. After the first high resolution fractionation via ion exchange LC, two distinct, well resolved zones were observed, both of
Discussion
We decided to investigate DH of a termite, because no DH was known from any species of order Isoptera. This order is regarded as phylogenetically more advanced than Blattodea (cockroaches, from which Peram-DP and Dippu-DH46 were identified) or Orthoptera (locusts and crickets, from which Locmi-DH and Achdo-DP, have been identified, respectively; see Fig. 3). The Orthoptera are ranked just below Blattodea by taxonomists. From the more highly evolved holometabolous orders, two DH are known from
Acknowledgements
This research was supported by NIH grant GM48172. We thank Dr. Barbara L. Thorne (U. of Maryland) for identifying our insects as Zootermopsis nevadensis subspecies nevadensis, and Dr. David Quilici for ESI-MS analysis of Zoone-DH and peptide fragments.
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Functional characterization of a diuretic hormone receptor associated with desiccation, starvation and temperature tolerance in gypsy moth, Lymantria dispar
2022, Pesticide Biochemistry and PhysiologyCitation Excerpt :Diuretic hormone (DH) regulates insect water and ion homeostasis, excretion and feeding behavior by binding to their paired receptors (Gäde and Goldsworthy, 2003, Schooley et al. 2012). DH was first identified in the moth, Manduca sexta (Kataoka et al., 1989), then DHs and its receptors have been identified in diverse insect species (Reagan, 1994; Furuya et al., 1998; Baldwin et al., 2001). The diuretic hormone receptor (DHR) is a family B1 (secretin) of GPCR (Lee et al., 2016), and is one of these neuropeptide receptors which regulates water and ion balance.
Can BRET-based biosensors be used to characterize G-protein mediated signaling pathways of an insect GPCR, the Schistocerca gregaria CRF-related diuretic hormone receptor?
2020, Insect Biochemistry and Molecular BiologyCitation Excerpt :While the physiological role of Schgr-DH was previously investigated in S. gregaria (Van Wielendaele et al., 2012), its receptor was never molecularly characterized. Moreover, this newly characterized receptor, Schgr-CFR-DHR, was also chosen as a proof-of-principle, since it is well-documented that CRF-DHs exert effects on their cellular targets through their interaction with receptors belonging to the secretin receptor-related GPCR family (family B), for which cAMP has been identified to act as an intracellular second messenger of CRF-DH in numerous insects, both in vivo and in vitro (Audsley et al., 1995; Baldwin et al., 2001; Clottens et al., 1994; Hector et al., 2009; Johnson et al., 2004; Lee et al., 2016; Reagan, 1996; Tobe et al., 2005). In addition, the production of primary urine is dependent on cAMP, which increases cationic transport (such as K+ and Na+) into the Malpighian tubules (Beyenbach, 1995; O'Donnell et al., 1996).
Water Homeostasis and Osmoregulation as Targets in the Control of Insect Pests
2013, Advances in Insect PhysiologyCitation Excerpt :Two CRF-like DH peptides, isolated from T. molitor (Tenmo-DH37 and Tenmo-DH47) (Furuya et al., 1995, 1998), are highly potent neurohormones stimulating diuresis in isolated MTs at EC50 values of 12 and 27 nM, respectively (Wiehart et al., 2002), and two similar DHs (DH37 and DH47) were isolated from the closely related T. castaneum species (Li et al., 2008). Other CRF-like DHs from a variety of insect species include the diuteric hormone Dippu-DH46 isolated from the Pacific beetle cockroach, Diploptera punctata (Furuya et al., 2000); Zoone-DH was reported from the dark Pacific Coast rottenwood termite, Zootermopsis nevadensis (Baldwin et al., 2001) and a diuretic peptide (Peram DH) was obtained from the American cockroach Periplaneta americana (Kay et al., 1992). A CRF-like factor with diuretic activity previously found in Rhodnius (Te Brugge et al., 2002) was later identified by using database sequences of the Rhodnius genome project (Te Brugge et al., 2011).
Anti-diuretic factors in insects: The role of CAPA peptides
2012, General and Comparative EndocrinologyCitation Excerpt :Factors that regulate the excretory system in insects include a variety of peptide families along with biogenic amines [9,10,16,21,31,70,76,102,104]. Diuretic factors that stimulate fluid secretion by the MTs include the biogenic amines tyramine [9,10] and serotonin (5-hydroxytryptamine; 5HT) [73,74], as well as several families of peptides such as the corticotropin-releasing factor (CRF)-related peptides [6,7,33,45,46,84], insect kinins [8,18,38,39,105,110], calcitonin-like peptides [17,34] and the CAPA family of peptides [24,25,47,91]. Generally speaking, few of these have been shown to be true diuretic hormones (i.e. actually shown to be present in the haemolymph at appropriate times).
Natriuresis and diuretic hormone synergism in R. prolixus upper Malpighian tubules is inhibited by the anti-diuretic hormone, RhoprCAPA-α2
2012, Journal of Insect PhysiologyCitation Excerpt :Tissues principally involved in maintaining ionic balance include the Malpighian (renal) tubules (MTs) and hindgut, which together form the functional kidney. Numerous diuretic factors have been identified in insects that belong to a variety of peptide families and also include at least two biogenic amines (Baldwin et al., 2001; Blackburn et al., 1991; Blumenthal, 2003; Coast et al., 2001, 2005; Furuya et al., 1995, 1998, 2000a,b; Kean et al., 2002; Maddrell et al., 1991; Orchard, 2006; Te Brugge et al., 2011b). Although less studied, anti-diuretic factors which act upon the hindgut to increase reabsorption of selected ions and water have also been identified (Audsley et al., 1992; Fournier and Girardie, 1988; Meredith et al., 1996; Phillips et al., 1980; Spring and Phillips, 1980).
Hormones Controlling Homeostasis in Insects
2012, Insect Endocrinology
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