scholarly journals A comparison of the effects of two putative diuretic hormones from Locusta migratoria on isolated locust malpighian tubules

1993 ◽  
Vol 175 (1) ◽  
pp. 1-14 ◽  
Author(s):  
G. M. Coast ◽  
R. C. Rayne ◽  
T. K. Hayes ◽  
A. I. Mallet ◽  
K. S. Thompson ◽  
...  

Previous work has shown that a peptide related to arginine vasopressin is present in the suboesophageal ganglion of the locust, Locusta migratoria. This peptide was determined to be an anti-parallel dimer of the nonapeptide Cys-Leu-Ile-Thr-Asn-Cys-Pro-Arg-Gly-NH2 and was reported to stimulate cyclic AMP production and fluid secretion in a combined Malpighian tubules and midgut preparation from locusts. For these reasons the peptide has been called the arginine-vasopressin-like insect diuretic hormone (AVP-like IDH). Recently, a second diuretic peptide (Locusta-DP), which is related to corticotropin releasing factor, has been identified: this is a potent stimulant of fluid secretion and cyclic AMP production by isolated locust tubules. Because water balance in insects is likely to be controlled by a cocktail of hormones acting on both Malpighian tubules and hindgut, this study directly compares the activity of these two peptides in fluid secretion and cyclic AMP production bioassays on one target organ, the isolated Malpighian tubule of Locusta migratoria. Locusta-DP was synthesised directly, whereas the dimeric AVP-like IDH was obtained by oxidation of a synthetic nonapeptide monomer. Products were separated by RP-HPLC and their structures unequivocally confirmed by enzymatic digestion, sequence analysis and electrospray mass spectrometry. We show that Locusta-DP causes strong stimulation of fluid secretion and cyclic AMP production, whereas the AVP-like IDH has no effect in either assay. These findings are discussed in the light of recent work on the anatomy and physiology of the vasopressin-like immunoreactive (VPLI) neurones in the suboesophageal ganglion of Locusta migratoria, the proposed source of the AVP-like peptide.

1993 ◽  
Vol 178 (1) ◽  
pp. 231-243 ◽  
Author(s):  
N. Audsley ◽  
G. M. Coast ◽  
D. A. Schooley

1. Manduca sexta diuretic hormone (Mas-DH) stimulates fluid secretion by adult Malpighian tubules of M. sexta, demonstrating its site of diuretic action in M. sexta for the first time. It was not possible to develop a suitable bioassay to measure fluid secretion in larval proximal tubules. 2. Mas-DH has an antidiuretic action on the cryptonephric complex of larval M. sexta because it increases fluid absorption from the rectum. It appears that in this complex Mas-DH is acting on a Na+/K+/2Cl- co-transporter, presumably on the basal membrane of the cryptonephric Malpighian tubules, because Mas-DH-stimulated fluid absorption by the cryptonephric complex is inhibited by bumetanide or the removal of Cl-, Na+ or K+ from the haemolymph side of the tissue. This is the first demonstration of hormonal control of fluid absorption by the cryptonephric complex. 3. Concomitant with the stimulation of fluid transport, Mas-DH increases the amount of cyclic AMP secreted by adult Malpighian tubules and the cryptonephric complex. In addition, Mas-DH promotes cyclic AMP production by the larval proximal tubules.


1997 ◽  
Vol 200 (17) ◽  
pp. 2363-2367 ◽  
Author(s):  
M C Quinlan ◽  
N J Tublitz ◽  
M J O'Donnell

Rhodnius prolixus eliminates NaCl-rich urine at high rates following its infrequent but massive blood meals. This diuresis involves stimulation of Malpighian tubule fluid secretion by diuretic hormones released in response to distention of the abdomen during feeding. The precipitous decline in urine flow that occurs several hours after feeding has been thought until now to result from a decline in diuretic hormone release. We suggest here that insect cardioacceleratory peptide 2b (CAP2b) and cyclic GMP are part of a novel mechanism of anti-diuresis. Secretion rates of 5-hydroxytryptamine-stimulated Malpighian tubules are reduced by low doses of CAP2b or cyclic GMP. Maximal secretion rates are restored by exposing tubules to 1 mmol l-1 cyclic AMP. Levels of cyclic GMP in isolated tubules increase in response to CAP2b, consistent with a role for cyclic GMP as an intracellular second messenger. Levels of cyclic GMP in tubules also increase as urine output rates decline in vivo, suggesting a physiological role for this nucleotide in the termination of diuresis.


1991 ◽  
Vol 158 (1) ◽  
pp. 165-180
Author(s):  
ANGELA WENNING ◽  
U. T. E. GREISINGER ◽  
JACQUES P. PROUX

Fluid secretion by isolated upper and lower portions of Malpighian tubules in the centipede Lithobius forficatus L. was studied. Ion requirements, cellular and transepithelial potentials, dependence on external osmolality and the effects of an insect diuretic factor and transport-active drugs were investigated. Unlike many insects, L. forficatus exhibited strongly Na+-dependent, K+-independent urine formation. However, as in many insects, upper and lower tubule portions from L. forficatus produced a K+-enriched, hypertonic fluid, and the transepithelial potential was positive with respect to the haemolymph. Furthermore, furosemide (5×10−4moll−1) reversibly inhibited urine formation. Ouabain, even at 10−3moll−1, had little effect on urine flow rate in upper tubules but inhibited secretion in lower tubules, albeit not completely. Locust diuretic hormone (at 10−7moll−1) enhanced fluid secretion in L. forficatus, but its action was not mimicked by dibutyryl cyclic AMP. The results suggest that some characteristics attributed exclusively to insects are common to non-insect arthropods.


2002 ◽  
Vol 205 (11) ◽  
pp. 1645-1655 ◽  
Author(s):  
Juan P. Ianowski ◽  
Robert J. Christensen ◽  
Michael J. O'Donnell

SUMMARYIntracellular ion activities (aion) and basolateral membrane potential (Vbl) were measured in Malpighian tubule cells of Rhodnius prolixus using double-barrelled ion-selective microelectrodes. In saline containing 103mmoll-1Na+, 6mmoll-1 K+ and 93mmoll-1Cl-, intracellular ion activities in unstimulated upper Malpighian tubules were 21, 86 and 32mmoll-1, respectively. In serotonin-stimulated tubules, aCl was unchanged, whereas aNa increased to 33mmoll-1 and aK declined to 71mmoll-1. Vbl was -59mV and -63mV for unstimulated and stimulated tubules, respectively. Calculated electrochemical potentials(Δμ/F) favour passive movement of Na+ into the cell and passive movement of Cl- out of the cell in both unstimulated and serotonin-stimulated tubules. Passive movement of K+ out of the cell is favoured in unstimulated tubules. In stimulated tubules, Δμ/F for K+ is close to 0 mV.The thermodynamic feasibilities of Na+-K+-2Cl-, Na+-Cl-and K+-Cl- cotransporters were evaluated by calculating the net electrochemical potential (Δμnet/F) for each transporter. Our results show that a Na+-K+-2Cl- or a Na+-Cl- cotransporter but not a K+-Cl- cotransporter would permit the movement of ions into the cell in stimulated tubules. The effects of Ba2+ and ouabain on Vbl and rates of fluid and ion secretion show that net entry of K+ through ion channels or the Na+/K+-ATPase can be ruled out in stimulated tubules. Maintenance of intracellular Cl- activity was dependent upon the presence of both Na+ and K+ in the bathing saline. Bumetanide reduced the fluxes of both Na+ and K+. Taken together, the results support the involvement of a basolateral Na+-K+-2Cl- cotransporter in serotonin-stimulated fluid secretion by Rhodnius prolixus Malpighian tubules.


Genetics ◽  
2020 ◽  
Vol 214 (2) ◽  
pp. 235-264 ◽  
Author(s):  
Erez Cohen ◽  
Jessica K. Sawyer ◽  
Nora G. Peterson ◽  
Julian A. T. Dow ◽  
Donald T. Fox

The insect excretory system contains two organ systems acting in concert: the Malpighian tubules and the hindgut perform essential roles in excretion and ionic and osmotic homeostasis. For over 350 years, these two organs have fascinated biologists as a model of organ structure and function. As part of a recent surge in interest, research on the Malpighian tubules and hindgut of Drosophila have uncovered important paradigms of organ physiology and development. Further, many human disease processes can be modeled in these organs. Here, focusing on discoveries in the past 10 years, we provide an overview of the anatomy and physiology of the Drosophila excretory system. We describe the major developmental events that build these organs during embryogenesis, remodel them during metamorphosis, and repair them following injury. Finally, we highlight the use of the Malpighian tubules and hindgut as accessible models of human disease biology. The Malpighian tubule is a particularly excellent model to study rapid fluid transport, neuroendocrine control of renal function, and modeling of numerous human renal conditions such as kidney stones, while the hindgut provides an outstanding model for processes such as the role of cell chirality in development, nonstem cell–based injury repair, cancer-promoting processes, and communication between the intestine and nervous system.


1999 ◽  
Vol 202 (15) ◽  
pp. 2017-2027 ◽  
Author(s):  
V.A. Te Brugge ◽  
S.M. Miksys ◽  
G.M. Coast ◽  
D.A. Schooley ◽  
I. Orchard

The blood-feeding bug Rhodnius prolixus ingests a large blood meal, and this is followed by a rapid diuresis to eliminate excess water and salt. Previous studies have demonstrated that serotonin and an unidentified peptide act as diuretic factors. In other insects, members of the corticotropin-releasing factor (CRF)-related peptide family have been shown to play a role in post-feeding diuresis. Using fluorescence immunohistochemistry and immunogold labelling with antibodies to the Locusta CRF-like diuretic hormone (Locusta-DH) and serotonin, we have mapped the distribution of neurones displaying these phenotypes in R. prolixus. Strong Locusta-DH-like immunoreactivity was found in numerous neurones of the central nervous system (CNS) and, in particular, in medial neurosecretory cells of the brain and in posterior lateral neurosecretory cells of the mesothoracic ganglionic mass (MTGM). Positively stained neurohaemal areas were found associated with the corpus cardiacum (CC) and on abdominal nerves 1 and 2. In addition, Locusta-DH-like immunoreactive nerve processes were found over the posterior midgut and hindgut. Double-labelling studies for Locusta-DH-like and serotonin-like immunoreactivity demonstrated some co-localisation in the CNS; however, no co-localisation was found in the medial neurosecretory cells of the brain, the posterior lateral neurosecretory cells of the MTGM or neurohaemal areas. To confirm the presence of a diuretic factor in the CC and abdominal nerves, extracts were tested in Malpighian tubule secretion assays and cyclic AMP assays. Extracts of the CC and abdominal nerves caused an increase in the rate of secretion and an increase in the level of cyclic AMP in the Malpighian tubules of fifth-instar R. prolixus. The presence of the peptide in neurohaemal terminals of the CC and abdominal nerves that are distinct from serotonin-containing terminals indicates that the peptide is capable of being released into the haemolymph and that this release can be independent of the release of serotonin.


1971 ◽  
Vol 54 (3) ◽  
pp. 779-804 ◽  
Author(s):  
S.H. P. MADDRELL ◽  
D.E. M. PILCHER ◽  
B.O. C. GARDINER

1. 5-Hydroxytryptamine (5-HT) and a few of its derivatives will cause an acceleration of secretion by the isolated Malpighian tubules of Rhodnius prolixus and Carausius morosus. Substitution in the terminal amino group of 5-HT causes little loss of effect, but changes in the indole ring and the hydroxy group in general abolish activity. 2. Most derivatives of 5-HT which do not stimulate secretion, and many other related amines, act as inhibitors of secretion in Rhodnius. This inhibition is probably of the competitive type. 3. Substances which stimulate secretion in Rhodnius are easily washed off the Malpighian tubules, although they will stimulate secretion at very low concentrations By contrast, competitive inhibitors wash away relatively slowly and inhibit only at much higher concentrations. It is suggested therefore that stimulation involves only momentary interactions between stimulant and receptor sites, but that competitive inhibitors spend considerably longer attached to the sites. 4. Cyclic 3',5'-adenosine monophosphate (cyclic AMP) will stimulate secretion by tubules of both Rhodnius and Carausius. Tubules of both species are very sensitive to cyclic AMP, measurable effects being produced at concentrations of 4 x 10-5 M/l in Rhodnius and 10-4 M/l in Carausius. 5. Aminophylline (theophylline ethylene diamine) at a concentration of 10-4 M/l stimulates secretion by Carausius tubules. This compound and other inhibitors of phosphodiesterase have no stimulatory action on Rhodnius tubules, but any such effect may be masked by the adverse reaction of the tubules to the high concentrations necessary. 6. Inhibitors of 5-HT action such as tryptamine and tyramine also inhibit secretion by Rhodnius Malpighian tubules induced by cyclic AMP. This may be due to a dependence of the action of cyclic AMP on the state of the diuretic hormone receptor site. 7. The evidence is consistent with the idea that 5-HT, other stimulant molecules related to 5-HT and the diuretic hormones interact with Malpighian tubule cells of Rhodnius and Carausius at specific sites, probably on the cell membrane facing the haemolymph. As a result of this, secretion is induced, possibly through the action of intracellular cyclic AMP produced as a response.


2002 ◽  
Vol 205 (4) ◽  
pp. 493-501 ◽  
Author(s):  
U. I. M. Wiehart ◽  
S. W. Nicolson ◽  
R. A. Eigenheer ◽  
D. A. Schooley

SUMMARYFluid 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-DH37 and Tenmo-DH47, were found to stimulate Tenebrio molitor tubules in vitro in a dose-dependent manner with EC50 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-DH37 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 CAP2b, originally isolated from Manduca sexta, also increases intracellular cyclic GMP levels and inhibited fluid secretion by Tenebrio molitor tubules, with an EC50 value of 85 nmol l–1. This inhibitory effect was reversed by Tenmo-DH37. 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.


2010 ◽  
Vol 298 (3) ◽  
pp. R642-R660 ◽  
Author(s):  
Peter M. Piermarini ◽  
Laura F. Grogan ◽  
Kenneth Lau ◽  
Li Wang ◽  
Klaus W. Beyenbach

Transepithelial fluid secretion across the renal (Malpighian) tubule epithelium of the mosquito ( Aedes aegypti ) is energized by the vacuolar-type (V-type) H+-ATPase and not the Na+-K+-ATPase. Located at the apical membrane of principal cells, the V-type H+-ATPase translocates protons from the cytoplasm to the tubule lumen. Secreted protons are likely to derive from metabolic H2CO3, which raises questions about the handling of HCO3−by principal cells. Accordingly, we tested the hypothesis that a Cl/HCO3anion exchanger (AE) related to the solute-linked carrier 4 (SLC4) superfamily mediates the extrusion of HCO3−across the basal membrane of principal cells. We began by cloning from Aedes Malpighian tubules a full-length cDNA encoding an SLC4-like AE, termed AeAE. When expressed heterologously in Xenopus oocytes, AeAE is both N- and O-glycosylated and mediates Na+-independent intracellular pH changes that are sensitive to extracellular Cl−concentration and to DIDS. In Aedes Malpighian tubules, AeAE is expressed as two distinct forms: one is O-glycosylated, and the other is N-glycosylated. Significantly, AeAE immunoreactivity localizes to the basal regions of stellate cells but not principal cells. Concentrations of DIDS that inhibit AeAE activity in Xenopus oocytes have no effects on the unstimulated rates of fluid secretion mediated by Malpighian tubules as measured by the Ramsay assay. However, in Malpighian tubules stimulated with kinin or calcitonin-like diuretic peptides, DIDS reduces the diuretic rates of fluid secretion to basal levels. In conclusion, Aedes Malpighian tubules express AeAE in the basal region of stellate cells, where this transporter may participate in producing diuretic rates of transepithelial fluid secretion.


1995 ◽  
Vol 198 (3) ◽  
pp. 793-804 ◽  
Author(s):  
M Patel ◽  
T Hayes ◽  
G Coast

Locusta-DP is a corticotropin-releasing factor (CRF)-related diuretic peptide isolated from the migratory locust Locusta migratoria. At nanomolar concentrations, synthetic Locusta-DP stimulated fluid secretion and cyclic AMP production by Malpighian tubules isolated in vitro and increased the rate of amaranth clearance in starved locusts to levels comparable with those observed during post-feeding diuresis. The peptide also caused a marked (approximately 10 %), but short-lived, reduction in the haemolymph volume of starved locusts. A polyclonal antiserum raised against Locusta-DP(29-46) was shown to block peptidergic signal transfer in vitro and in vivo. Pre-treatment of Locusta-DP (5 nmol l-1) with antiserum diluted 1:100 resulted in a rapid reduction in the free peptide concentration to less than 1 nmol l-1, the threshold for a measurable effect on cyclic AMP production by isolated tubules. In intact insects, passive immunization with Locusta-DP antiserum blocked increases in the rate of amaranth clearance in response to exogenous diuretic peptide or in response to feeding. The latter was due specifically to the binding of Locusta-DP, because when the relevant antibodies were preadsorbed with Locusta-DP(29-46), the antiserum had no effect on amaranth clearance by recently fed insects. This provides unequivocal evidence of a hormonal function for Locusta-DP in the control of primary urine production.


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