Original ArticlesIsolation and Identification of a Second Diuretic Hormone from Tenebrio molitor 1
Section snippets
Experimental Animals
T. molitor were reared as described earlier [14]. Heads of pupae were collected and stored at −80°C as described [14].
Bioassay
DH activity in fractions from all purification steps was detected by the elevation of cAMP in isolated Mt dissected from newly emerged adult T. molitor as described earlier [14]. Preparation of aliquots of fractions for bioassay was also performed as already described [14].
Extraction and Preliminary Purification (Step 1)
Collected heads (fresh weight ≈103 g) from ≈20,000 T. molitor pupae were homogenized in cold methylene
Extraction and Preliminary Purification (Step 1)
The preliminary purification used a Vydac C4 reversed-phase cartridge as already described for identifying T. molitor DH37 [14]. Fractions were assayed for the presence of a DH based on their ability to stimulate cAMP production in Mt of T. molitor. Strong diuretic activity was found in the fraction that eluted from the cartridge with 45% acetonitrile.
Liquid Chromatographic Purification (Steps 2 to 9)
The 45% CH3CN fraction from above was diluted with 12 volumes of 0.1% TFA and then loaded onto a Vydac C4 semipreparative column (Step 2)
Discussion
We report the identity of a second DH in head extracts of the coleopteran insect T. molitor. We named this hormone T. molitor DH47; the subscripted number signifies that it is a 47 amino acid peptide. Earlier, we identified a smaller DH from this species that we now designate T. molitor DH37 for consistency (rather than Tem-DH1). Both of these DH are members of a family of peptides that have sequence similarity to the sauvagine-CRF-urotensin I family of peptides from vertebrates (Fig. 2).
Acknowledgements
We thank Hewlett–Packard for sequence analysis of the N-terminal 40 residues of T. molitor DH47, Chris Grimley of Applied Biosystems for amino acid sequencing of the C-terminal fragment, Dr. Arnold Falick of PerSeptive Biosystems for the MALDI MS analysis of the C-terminal fragment, and Dr. George C. J. Fernandez for statistical analysis of the dose/response data. This work was supported by NIH #GM48172, and is a contribution of the Nevada Agriculture Experiment Station. We thank Dr. Houle Wang
References (39)
- et al.
Quantification of Locusta diuretic hormone in the central nervous system and corpora cardiacaInfluence of age and feeding status, and mechanism of release
Regul. Pept.
(1997) - et al.
Cross reactivity studies of CRF-related peptides on insect Malpighian tubules
Comp. Biochem. Physiol. [A]
(1995) - et al.
Serotonin-induced elevation of cAMP levels in the epidermis of the blood-sucking bug, Rhodnius prolixus
J. Insect Physiol.
(1990) - et al.
Isolation and identification of a new diuretic peptide from the tobacco hornworm, Manduca sexta
Biochem. Biophys. Res. Commun.
(1991) The excretory systemStructure and physiology
- et al.
Isolation and characterization of a diuretic peptide common to the house fly and stable fly
Peptides
(1994) Neuropeptides implicated in the control of diuresis in insects
Peptides.
(1996)- et al.
The diuretic activity of a series of cephalomyotropic neuropeptides, the achetakinins, on isolated Malpighian tubules of the house cricket, Acheta domesticus
J. Insect Physiol.
(1990) - et al.
Leucokinins, a new family of ion transport stimulators and inhibitors in insect Malpighian tubules
Life Sci.
(1989) - et al.
Isolation, primary structure and synthesis of leucokinins VII and VIIIThe final members of this new family of cephalomyotropic peptides isolated from head extracts of Leucophaea maderae
Comp. Biochem. Physiol.
(1987)
Isolation, characterization and biological activity of a CRF-related diuretic peptide from Periplaneta americana L
Regul. Pept.
Identification of a diuretic hormone of Locusta migratoria
Biochem. Biophys. Res. Commun.
Cyclic AMP in the Malpighian tubule fluid and in the urine of Rhodnius prolixus
Gen. Comp. Endocrinol.
5-Hydroxytryptamine stimulates fluid secretion in locust Malpighian tubules independently of cAMP
Comp. Biochem. Physiol.
Effects of biogenic amines and hormones on butterfly Malpighian tubulesdopamine stimulates fluid secretion
J. Insect Physiol.
Expression cloning of an insect diuretic hormone receptor—a member of the calcitonin/secretin receptor family
J. Biol. Chem.
Effects of 5-hydroxytryptamine on the Malpighian tubules of Aedes aegypti
J. Insect Physiol.
Assay and characterisation of diuretic factors in insects
J. Insect Physiol.
The effects of Manduca sexta diuretic hormone on fluid transport by the Malpighian tubules and cryptonephric complex of Manduca sexta
J. Exp. Biol.
Cited by (41)
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.
Receptors for neuronal or endocrine signalling molecules as potential targets for the control of insect pests
2014, Advances in Insect PhysiologyCitation Excerpt :In many insect species, diuretic peptides were discovered which showed a high resemblance to vertebrate corticotrophin-releasing factor (CRF), urotensin and urocortin (Gäde, 2004). Such CRF/DH, also referred to as DH44, were first identified in M. sexta (Kataoka et al., 1989b) and to date their occurrence has been confirmed in a wide variety of insect pests including the mealworm beetle Tenebrio molitor (Furuya et al., 1995, 1998), R. prolixus (Te Brugge et al., 2011), the whitelined sphinx moth Hyles lineata (Furuya et al., 2000b), P. americana (Kay et al., 1992), and the locusts L. migratoria (Kay et al., 1991; Lehmberg et al., 1991) and S. gregaria (Van Wielendaele et al., 2012). Furthermore, a gene encoding CRF/DH has also been found in the genome of An.
Water Homeostasis and Osmoregulation as Targets in the Control of Insect Pests
2013, Advances in Insect PhysiologyCitation Excerpt :Identical peptide sequences were isolated and identified in the house fly Musca domestica and the stable fly Stomoxys calcitrans (Clottens et al., 1994), and an almost identical sequence of the above dipterans was described for D. melanogaster (Drome-DH44) neuropeptide (Cabrero et al., 2002). 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).
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 EndocrinologyToward a consensus nomenclature for insect neuropeptides and peptide hormones
2011, PeptidesCitation Excerpt :Corticotropin releasing-factor (CRF)-related diuretic hormone (CRF-related DH): Members of this peptide family have diuretic activity in that they stimulate MT secretion, and have been described as both CRF-like and CRF-related, although the latter is more accurate as they are sufficiently similar to vertebrate CRF/urotensin I/urocortin/sauvagine to be placed in the same peptide superfamily [21]. Most insects appear to have a single isoform, but two isoforms are present in M. sexta [10,61], T. molitor [32,33], T. castaneum [69], and three isoforms in B. mori [103]. In T. castaneum and B. mori the isoforms are produced by alternate splicing of the gene [69,103], but in vertebrates the different paralogs of the CRF superfamily are each synthesized as unique prepropeptides, which are not splice variants.
- 1
Taken in part from a paper presented at a Joint Meeting of the European Neuropeptide Club-Winter Discussion and the Winter Neuropeptide Conference February 8–11, 1997, Kitzbühel, Austria.
- 2
Present address: Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080.