scholarly journals Ecdysone signaling is required for proper organization and fluid secretion of stellate cells in the Malpighian tubules of Drosophila melanogaster

2010 ◽  
Vol 54 (4) ◽  
pp. 635-642 ◽  
Author(s):  
Naveen-Kumar Gautam ◽  
Madhu Tapadia
Keyword(s):  
Drosophila Melanogaster ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Ecdysone Signaling

The malpighian tubules of Drosophila melanogaster: a novel phenotype for studies of fluid secretion and its control.

1994 ◽  
Vol 197 (1) ◽  
pp. 421-428 ◽  
Author(s):  
J A Dow ◽  
S H Maddrell ◽  
A Görtz ◽  
N J Skaer ◽  
S Brogan ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Fluid Secretion ◽  
Malpighian Tubules

scholarly journals A SLC4-like anion exchanger from renal tubules of the mosquito (Aedes aegypti): evidence for a novel role of stellate cells in diuretic fluid secretion

2010 ◽  
Vol 298 (3) ◽  
pp. R642-R660 ◽  
Author(s):  
Peter M. Piermarini ◽  
Laura F. Grogan ◽  
Kenneth Lau ◽  
Li Wang ◽  
Klaus W. Beyenbach
Keyword(s):  
Aedes Aegypti ◽  
Anion Exchanger ◽  
Xenopus Oocytes ◽  
Malpighian Tubule ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Renal Tubules ◽  
Basal Region ◽  
Principal Cells

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.

scholarly journals Isolation and characterization of a leucokinin-like peptide of Drosophila melanogaster

1999 ◽  
Vol 202 (24) ◽  
pp. 3667-3676 ◽  
Author(s):  
S. Terhzaz ◽  
F.C. O'Connell ◽  
V.P. Pollock ◽  
L. Kean ◽  
S.A. Davies ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Genomic Sequence ◽  
Genetic Model ◽  
Stellate Cells ◽  
Malpighian Tubules ◽  
Peptide Sequence ◽  
Renal Tubules ◽  
Isolation And Characterization ◽  
Wide Range ◽  
Intracellular Ca

The leucokinin (LK) family of neuropeptides has been found widely amongst invertebrates. A member of this family was purified from adults of the fruit fly Drosophila melanogaster. The peptide sequence for Drosophila leucokinin (DLK) was determined as Asn-Ser-Val-Val-Leu-Gly-Lys-Lys-Gln-Arg-Phe-His-Ser-Trp-Gly-amide, making it the longest member of the family characterized to date. Synthetic DLK peptide was shown to act to stimulate fluid secretion in D. melanogaster Malpighian (renal) tubules by approximately threefold, with an EC(50) of approximately 10(−)(10)mol l(−)(1), and a secondary effect at approximately 10(−)(7)mol l(−)(1). DLK also acted to elevate intracellular [Ca(2+)] in the Malpighian tubules by approximately threefold, with an EC(50) of 10(−)(10) to 10(−)(9)mol l(−)(1). Responses were detected in stellate cells and occasionally in principal cells, although at no concentration tested did [Ca(2+)] in the principal cell increase significantly above background. In stellate cells, DLK produced a biphasic rise in intracellular [Ca(2+)] from resting levels of 80–100 nmol l(−)(1), with a transient peak being followed by a slower rise that peaked at 200–300 nmol l(−)(1) after 3 s, then decayed over approximately 10 s. The wide range of concentrations over which DLK acts suggests the involvement of more than one receptor. The genomic sequence encoding the DLK peptide has been identified, and the gene has been named pp. The gene resides at cytological location 70E3-70F4 of chromosome 3L. The localisation of this first Drosophila LK gene in a genetic model permits a genetic analysis of the locus.

Neuropeptide stimulation of the nitric oxide signaling pathway in Drosophila melanogaster Malpighian tubules

1997 ◽  
Vol 273 (2) ◽  
pp. R823-R827 ◽  
Author(s):  
S. A. Davies ◽  
E. J. Stewart ◽  
G. R. Huesmann ◽  
N. J. Skaer ◽  
S. H. Maddrell ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Drosophila Melanogaster ◽  
Signaling Pathway ◽  
Guanylate Cyclase ◽  
Soluble Guanylate Cyclase ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
No Production ◽  
Intracellular Cgmp ◽  
Stimulation Of

Activation of the nitric oxide (NO) and guanosine 3', 5'-cyclic monophosphate (cGMP) signaling pathway stimulates fluid secretion by the Drosophila melanogaster Malpighian tubule. The neuropeptide cardioacceleratory peptide 2b (CAP2b) has been previously shown to stimulate fluid secretion in this epithelium by elevating intracellular cGMP levels. Therefore, it was of interest to investigate if CAP2b acts through NO in isolated tubules and thus presumably through stimulation of a tubule NO synthase (NOS). We show here by reverse-transcription polymerase chain reaction that Drosophila NOS (dNOS) is expressed in Malpighian tubules. Biochemical assays of NOS activity in whole tubules show that CAP2b significantly stimulates NOS activity. Additionally, fluid secretion and cyclic nucleotide assays show that CAP2b-induced elevation of intracellular cGMP levels and fluid secretion rates are dependent on the activation of a soluble guanylate cyclase. Treatment of tubules with a specific NOS inhibitor abolishes the CAP2b-induced rise in intracellular cGMP levels. These data indicate that CAP2b stimulates NOS and therefore, endogenous NO production, which, in turn, stimulates a soluble guanylate cyclase. This is the first demonstration of stimulation of an endogenous NOS by a defined peptide in Drosophila.

scholarly journals When two cells are better than one: specialized stellate cells provide a privileged route for uniquely rapid water flux in Drosophila renal tubule

2019 ◽  
Author(s):  
Pablo Cabrero ◽  
Selim Terhzaz ◽  
Anthony J. Dornan ◽  
Saurav Ghimire ◽  
Heather L. Holmes ◽  
...  
Keyword(s):  
Water Permeability ◽  
Plasma Membranes ◽  
Water Flux ◽  
Malpighian Tubule ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Principal Cells ◽  
Very High

AbstractInsects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 members of the Major Intrinsic Protein family are expressed at very high level in the fly renal tissue; the aquaporins Drip and Prip, and the aquaglyceroporins Eglp2 and Eglp4. As predicted from their structure and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes impacts tubule performance resulting in impaired hormone-induced fluid secretion. The Drosophila tubule has two main secretory cell types: active cation-transporting principal cells with the aquaglyceroporins localize to opposite plasma membranes and small stellate cells, the site of the chloride shunt conductance, with these aquaporins localising to opposite plasma membranes. This suggests a model in which cations are pumped by the principal cells, causing chloride to follow through the stellate cells in order to balance the charge. As a consequence, osmotically obliged water follows through the stellate cells. Consistent with this model, fluorescently labelled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects.Significance statementThe tiny insect renal (Malpighian) tubule can transport fluid at unparalleled speed, suggesting unique specialisations. Here we show that strategic allocation of Major Intrinsic Proteins (MIPs) to specific cells within the polarized tubule allow the separation of metabolically intense active cation transport from chloride and water conductance. This body plan is general to at least many higher insects, providing a clue to the unique success of the class Insecta.

Fluid secretion by isolated Malpighian tubules of Drosophila melanogaster Meig.: effects of organic anions, quinacrine and a diuretic factor found in the secreted fluid

1999 ◽  
Vol 202 (17) ◽  
pp. 2339-2348 ◽  
Author(s):  
J.A. Riegel ◽  
R.W. Farndale ◽  
S.H. Maddrell
Keyword(s):  
Drosophila Melanogaster ◽  
Adenylate Cyclase ◽  
Cyclic Nucleotides ◽  
Fluid Secretion ◽  
Organic Anions ◽  
Malpighian Tubules ◽  
Cyclase Activity ◽  
Adenylate Cyclase Activity ◽  
Phenol Red ◽  
Stimulation Of

Para-aminohippuric acid (PAH, 0.2 and 1 mmol l(−)(1)) had no effect on the basal fluid secretion rate (FSR) of isolated Malpighian tubules of Drosophila melanogaster Meig. and did not affect stimulation of the FSR induced by adenosine 3′,5′-monophosphate (cAMP). Phenol Red (phenolsulphonphthalein, PSP; 0.5 and 1 mmol l(−)(1)) slowed the FSR and abolished stimulation of the FSR by cAMP. Diodrast (1 mmol l(−)(1)) slightly, but significantly, reduced the FSR and greatly reduced the stimulation of the FSR normally provoked by cAMP and by the 3′,5′-monophosphates of guanosine (cGMP), inosine (cIMP) and uridine (cUMP). However, stimulation of the FSR by the 3′, 5′-monophosphate of cytidine (cCMP) was little affected by diodrast. Probenecid (0.2 or 1 mmol l(−)(1)) consistently stimulated the FSR, on average by approximately 25 %, but did not markedly inhibit the subsequent stimulation of the FSR by cAMP, cGMP or cIMP. However, the FSR of tubules stimulated by cGMP was temporarily lowered by probenecid. Quinacrine (0.1 mmol l(−)(1)) slowed basal FSR by an average of approximately 30 %, but subsequent stimulation of the FSR by cAMP was not noticeably affected. Both 0.1 mmol l(−)(1) cAMP and 1 mmol l(−)(1) probenecid stimulated adenylate cyclase activity in extracts of Malpighian tubules, but cIMP, cGMP, cUMP and diodrast were without effect in this regard. Uptake of radioactivity from a solution containing 500 nmol l(−)(1) [(3)H]cAMP and 9.5 μmol l(−)(1) cAMP was reduced by more than 90 % by 1 mmol l(−)(1) PSP, by approximately 40 % by 0.2 mmol l(−)(1) probenecid, by 36 % by 1 mmol l(−)(1) diodrast and by 30 % by 1 mmol l(−)(1) PAH. Neither 0.01 mmol l(−)(1) ouabain nor 0.1 mmol l(−)(1) quinacrine affected the uptake of [(3)H]cAMP by the Malpighian tubules. Fluid secreted by isolated Malpighian tubules of Drosophila melanogaster contains a factor that stimulated the FSR on average by approximately 50 %. The presence in the secreted fluid of cGMP at a concentration of 8.3 μmol l(−)(1) did not explain the stimulatory effect on FSR. These results support the existence of a carrier-mediated uptake of cyclic nucleotides into the Malpighian tubules of Drosophila melanogaster, possibly involving a multispecific transporter.

Contributions of K+:Cl- cotransport and Na+/K+-ATPase to basolateral ion transport in malpighian tubules of Drosophila melanogaster

1999 ◽  
Vol 202 (11) ◽  
pp. 1561-1570 ◽  
Author(s):  
S.M. Linton ◽  
M.J. O'Donnell
Keyword(s):  
Drosophila Melanogaster ◽  
Membrane Potential ◽  
Cyclic Amp ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Minor Role ◽  
Electrochemical Gradient ◽  
Tubule Cells

Mechanisms of Na+ and K+ transport across the basolateral membrane of isolated Malpighian tubules of Drosophila melanogaster were studied by examining the effects of ion substitution and putative inhibitors of specific ion transporters on fluid secretion rates, basolateral membrane potential and secreted fluid cation composition. Inhibition of fluid secretion by [(dihydroindenyl)oxy]alkanoic acid (DIOA) and bumetanide (10(−)4 mol l-1) suggested that a K+:Cl- cotransporter is the main route for K+ entry into the principal cells of the tubules. Differences in the effects of bumetanide on fluxes of K+ and Na+ are inconsistent with effects upon a basolateral Na+:K+:2Cl- cotransporter. Large differences in electrical potential across apical (>100 mV, lumen positive) and basolateral (<60 mV, cell negative) cell membranes suggest that a favourable electrochemical gradient for Cl- entry into the cell may be used to drive K+ into the cell against its electrochemical gradient, via a DIOA-sensitive K+:Cl- cotransporter. A Na+/K+-ATPase was also present in the basolateral membrane of the Malpighian tubules. Addition of 10(−)5 to 10(−)3 mol l-1 ouabain to unstimulated tubules depolarized the basolateral potential, increased the Na+ concentration of the secreted fluid by 50–73 % and increased the fluid secretion rate by 10–19 %, consistent with an increased availability of intracellular Na+. We suggest that an apical vacuolar-type H+-ATPase and a basolateral Na+/K+-ATPase are both stimulated by cyclic AMP. In cyclic-AMP-stimulated tubules, K+ entry is stimulated by the increase in the apical membrane potential, which drives K+:Cl- cotransport at a faster rate, and by the stimulation of the Na+/K+-ATPase. Fluid secretion by cyclic-AMP-stimulated tubules was reduced by 26 % in the presence of ouabain, suggesting that the Na+/K+-ATPase plays a minor role in K+ entry into the tubule cells. Malpighian tubules secreted a Na+-rich (150 mmol l-1) fluid at high rates when bathed in K+-free amino-acid-replete saline (AARS). Secretion in K+-free AARS was inhibited by amiloride and bafilomycin A1, but not by bumetanide or hydrochlorothiazide, which inhibit Na+:Cl- cotransport. There was no evidence for a Na+ conductance in the basolateral membrane of unstimulated or cyclic-AMP-stimulated tubules. Possible mechanisms of Na+ entry into the tubule cells include cotransport with organic solutes such as amino acids and glucose.

Stimulation of fluid secretion of malpighian tubules of drosophila melanogaster meig. by cyclic nucleotides of inosine, cytidine, thymidine and uridine

1998 ◽  
Vol 201 (24) ◽  
pp. 3411-3418
Author(s):  
J. A. Riegel ◽  
S. H. P. Maddrell ◽  
R. W. Farndale ◽  
F. M. Caldwell
Keyword(s):  
Drosophila Melanogaster ◽  
Cyclic Nucleotides ◽  
Cyclic Nucleotide ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Secretion Rate ◽  
External Application ◽  
Tubule Cells ◽  
Stimulation Of

External application of the 3',5'-cyclic monophosphates of inosine,cytidine, uridine and thymidine stimulated the fluid secretion rate (FSR)of Malpighian tubules isolated from Drosophila melanogaster. The evidence suggested that the cyclic nucleotides acted intracellularly in some capacity. Receptors of the 'purinergic' type appeared not to be major contributors to fluid secretion; of three purinergic agonists tried,adenosine, adenosine 5'-monophosphate (AMP) and adenosine 5'-triphosphate(ATP), only adenosine had an effect, but this was not observed consistently. None of the purinergic agonists interfered with the stimulation of the FSR by adenosine 3',5'-cyclic monophosphate (cAMP). The maximum stimulation of the fluid-secretion rate by any cyclic nucleotide was approximately double the unstimulated (control) rate. Tubules stimulated to less than maximal FSR by one cyclic nucleotide could be stimulated maximally by an appropriate concentration of another cyclic nucleotide. Malpighian tubules bathed in solutions that contained either[3H]cAMP or [3H]cGMP accumulated radioactivity to a level many times that in the medium. Accumulation of radioactivity by tubules bathed in 430 nmol l-1 [3H]cAMP was suppressed by 1 mmol l-1 non-radioactive cyclic nucleotides in the order cAMP>>cGMP>cIMP>cCMP; neither cTMP nor cUMP suppressed the accumulation of [3H]cAMP. Approximately 35 % of the[3H]cAMP and 80 % of the [3H]cGMP that entered the Malpighian tubule cells was metabolised to compounds that were not identified. It was concluded that cyclic nucleotides enter the Malpighian tubule cells by at least one transport mechanism which is particularly sensitive to purine-based nucleotides.

scholarly journals Specialized stellate cells offer a privileged route for rapid water flux in Drosophila renal tubule

2020 ◽  
Vol 117 (3) ◽  
pp. 1779-1787 ◽  
Author(s):  
Pablo Cabrero ◽  
Selim Terhzaz ◽  
Anthony J. Dornan ◽  
Saurav Ghimire ◽  
Heather L. Holmes ◽  
...  
Keyword(s):  
Water Permeability ◽  
Plasma Membranes ◽  
Water Flux ◽  
Stellate Cells ◽  
Fluid Secretion ◽  
Cell Types ◽  
Renal Tissue ◽  
Malpighian Tubules ◽  
Very High

Insects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 genes of the major intrinsic protein family are expressed at a very high level in the fly renal tissue: the aquaporins (AQPs) Drip and Prip and the aquaglyceroporins Eglp2 and Eglp4. As predicted from their structure, and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip, and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes result in impaired hormone-induced fluid secretion. The Drosophila tubule has 2 main secretory cell types: active cation-transporting principal cells, wherein the aquaglyceroporins localize to opposite plasma membranes, and small stellate cells, the site of the chloride shunt conductance, with these AQPs localizing to opposite plasma membranes. This suggests a model in which osmotically obliged water flows through the stellate cells. Consistent with this model, fluorescently labeled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects in regulating their internal environments.

Sign in / Sign up

Export Citation Format

Share Document