scholarly journals A novel role for a Drosophila homologue of cGMP-specific phosphodiesterase in the active transport of cGMP

2005 ◽  
Vol 393 (2) ◽  
pp. 481-488 ◽  
Author(s):  
Jonathan P. Day ◽  
Miles D. Houslay ◽  
Shireen-A. Davies
Keyword(s):  
Active Transport ◽  
Transgenic Animals ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Visual Signal ◽  
Type I ◽  
Direct Role ◽  
Principal Cells ◽  
Targeted Expression ◽  
Apical Membranes

cGMP was first discovered in urine, demonstrating that kidney cells extrude this cyclic nucleotide. Drosophila Malpighian tubules provide a model renal system in which a homologue of mammalian PDE (phosphodiesterase) 6 is expressed. In humans, this cG-PDE (cGMP-specific PDE) is specifically expressed in the retinal system, where it controls visual signal transduction. In order to gain insight into the functional role of DmPDE6 (Drosophila PDE6-like enzyme) in epithelial function, we generated transgenic animals with targeted expression of DmPDE6 to tubule Type I (principal) cells. This revealed localization of DmPDE6 primarily at the apical membranes. As expected, overexpression of DmPDE6 resulted in elevated cG-PDE activity and decreased tubule cGMP content. However, such targeted overexpression of DmPDE6 creates a novel phenotype that manifests itself in inhibition of the active transport and efflux of cGMP by tubules. This effect is specific to DmPDE6 action, as no effect on cGMP transport is observed in tubules from a bovine PDE5 transgenic line which display reduced rates of fluid secretion, an effect not seen in DmPDE6 transgenic animals. Specific ablation of DmPDE6 in tubule principal cells, via expression of a targeted DmPDE6 RNAi (RNA interference) transgene, conferred increased active transport of cGMP, confirming a direct role for DmPDE6 in regulating cGMP transport in tubule principal cells. Pharmacological inhibition of DmPDE6 in wild-type tubules using the cG-PDE inhibitor, zaprinast, similarly results in stimulated cGMP transport. We provide the first demonstration of a novel role for a cG-PDE in modulating cGMP transport and efflux.

Membrane dynamics in insect malpighian tubules

1989 ◽  
Vol 257 (5) ◽  
pp. R967-R972
Author(s):  
T. J. Bradley
Keyword(s):  
Ion Transport ◽  
Driving Force ◽  
Fluid Secretion ◽  
Membrane Dynamics ◽  
Malpighian Tubules ◽  
Membrane Insertion ◽  
Cation Pump ◽  
Active Ion Transport ◽  
Urine Formation ◽  
Apical Membranes

Urine formation in insects occurs in the Malpighian tubules by means of active ion transport and osmotically coupled water flow. The rates of urine formation can vary with time and can be modulated by diuretic hormones, developmental events, and intracellular parasitism. This paper reviews a number of recent studies in which it has been demonstrated that variations in transport rate are associated with substantial changes in tubule ultrastructure in the form of membrane insertion into and deletion from the apical microvilli. The principal driving force for fluid movement in Malpighian tubules is thought to be a common cation pump located in the apical membranes. It is proposed that modulation of the apical microvillar membrane may reflect regulation by the cells of the number of common cation pump units involved in fluid secretion.

Active Transport by Insect Malpighian Tubules of Acidic Dyes and of Acylamides

1974 ◽  
Vol 61 (2) ◽  
pp. 357-377 ◽  
Author(s):  
S. H. P. MADDRELL ◽  
B. O. C. GARDINER ◽  
D. E. M. PILCHER ◽  
S. E. REYNOLDS
Keyword(s):  
Active Transport ◽  
Organic Anion ◽  
Indigo Carmine ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Passive Permeability ◽  
Dye Transport ◽  
Tubule Lumen ◽  
Conjugated Compounds ◽  
Tubule Wall

Insect Malpighian tubules carry out active transport of two types of organic anion: acylamides (such as p-aminohippuric acid) and sulphonates (such as indigo carmine and amaranth). There are separate mechanisms for the transport of these two classes of compounds. The degree to which these compounds are concentrated depends critically on the passive permeability of the tubule wall. In the permeable Malpighian tubules of Calliphora, small transported molecules readily escape from the tubule lumen. At low rates of fluid secretion the net rate of dye transport is thereby very much reduced. As a result the rate of dye transport in this insect depends on the rate of fluid secretion, although the processes are not rigidly linked. In the less permeable tubules of Rhodnius and Carausius, dye secretion is not affected by the rate of fluid secretion. The active transport of these two types of compounds is a means of clearing from the haemolymph the conjugated compounds which are the products of detoxication of potentially toxic products of metabolism.

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 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.

scholarly journals Role of an apical K,Cl cotransporter in urine formation by renal tubules of the yellow fever mosquito (Aedes aegypti)

2011 ◽  
Vol 301 (5) ◽  
pp. R1318-R1337 ◽  
Author(s):  
Peter M. Piermarini ◽  
Rebecca M. Hine ◽  
Matthew Schepel ◽  
Jeremy Miyauchi ◽  
Klaus W. Beyenbach
Keyword(s):  
Aedes Aegypti ◽  
Epithelial Transport ◽  
Input Resistance ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Cell Swelling ◽  
Renal Tubules ◽  
Transport Pathway ◽  
Principal Cells

The K,Cl cotransporters (KCCs) of the SLC12 superfamily play critical roles in the regulation of cell volume, concentrations of intracellular Cl−, and epithelial transport in vertebrate tissues. To date, the role(s) of KCCs in the renal functions of mosquitoes and other insects is less clear. In the present study, we sought molecular and functional evidence for the presence of a KCC in renal (Malpighian) tubules of the mosquito Aedes aegypti . Using RT-PCR on Aedes Malpighian tubules, we identified five alternatively spliced partial cDNAs that encode putative SLC12-like KCCs. The majority transcript is AeKCC1-A1; its full-length cDNA was cloned. After expression of the AeKCC1-A protein in Xenopus oocytes, the Cl−-dependent uptake of 86Rb+ is 1) activated by 1 mM N-ethylmaleimide and cell swelling, 2) blocked by 100 μM dihydroindenyloxyalkanoic acid (DIOA), and 3) dependent upon N-glycosylation of AeKCC1-A. In Aedes Malpighian tubules, AeKCC1 immunoreactivity localizes to the apical brush border of principal cells, which are the predominant cell type in the epithelium. In vitro physiological assays of Malpighian tubules show that peritubular DIOA (10 μM): 1) significantly reduces both the control and diuretic rates of transepithelial fluid secretion and 2) has negligible effects on the membrane voltage and input resistance of principal cells. Taken together, the above observations indicate the presence of a KCC in the apical membrane of principal cells where it participates in a major electroneutral transport pathway for the transepithelial secretion of fluid in this highly electrogenic epithelium.

Dibutyryl cAMP activates bumetanide-sensitive electrolyte transport in Malpighian tubules

1991 ◽  
Vol 261 (3) ◽  
pp. C521-C529 ◽  
Author(s):  
J. L. Hegarty ◽  
B. Zhang ◽  
T. L. Pannabecker ◽  
D. H. Petzel ◽  
M. D. Baustian ◽  
...  
Keyword(s):  
Yellow Fever ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Fluid Secretion ◽  
Ringer Solution ◽  
Malpighian Tubules ◽  
Membrane Voltage ◽  
Dibutyryl Camp ◽  
K Secretion ◽  
Transepithelial Voltage

The effects of dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) and bumetanide (both 10(-4) M) on transepithelial Na+, K+, Cl-, and fluid secretion and on tubule electrophysiology were studied in isolated Malpighian tubules of the yellow fever mosquito Aedes aegypti. Peritubular DBcAMP significantly increased Na+, Cl-, and fluid secretion but decreased K+ secretion. In DBcAMP-stimulated tubules, bumetanide caused Na+, Cl-, and fluid secretion to return to pre-cAMP control rates and K+ secretion to decrease further. Peritubular bumetanide significantly increased Na+ secretion and decreased K+ secretion so that Cl- and fluid secretion did not change. In bumetanide-treated tubules, the secretagogue effects of DBcAMP are blocked. In isolated Malpighian tubules perfused with symmetrical Ringer solution, DBcAMP significantly hyperpolarized the transepithelial voltage (VT) and depolarized the basolateral membrane voltage (Vbl) with no effect on apical membrane voltage (Va). Total transepithelial resistance (RT) and the fractional resistance of the basolateral membrane (fRbl) significantly decreased. Bumetanide also hyperpolarized VT and depolarized Vbl, however without significantly affecting RT and fRbl. Together these results suggest that, in addition to stimulating electroconductive transport, DBcAMP also activates a nonconductive bumetanide-sensitive transport system in Aedes Malpighian tubules.

A cellular pathway for Cl− during fluid secretion in ant Malpighian tubules: Evidence from ion-sensitive microelectrode studies?

1995 ◽  
Vol 41 (8) ◽  
pp. 695-703 ◽  
Author(s):  
S. Dijkstra ◽  
A. Leyssens ◽  
E. Van Kerkhove ◽  
W. Zeiske ◽  
P. Steels
Keyword(s):  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Cellular Pathway

Chloride channels in apical membrane patches of stellate cells of Malpighian tubules of Aedes aegypti

2001 ◽  
Vol 204 (2) ◽  
pp. 367-378 ◽  
Author(s):  
K.R. O'Connor ◽  
K.W. Beyenbach
Keyword(s):  
Aedes Aegypti ◽  
Apical Membrane ◽  
Stellate Cells ◽  
Malpighian Tubules ◽  
Type I ◽  
Open Probability ◽  
Cl Secretion ◽  
Cl Channel ◽  
Open Time ◽  
Channel Type

Stellate cells of Aedes aegypti Malpighian tubules were investigated using patch-clamp methods to probe the route of transepithelial Cl(−) secretion. Two types of Cl(−) channel were identified in excised, inside-out apical membrane patches. The first Cl(−) channel, type I, had a conductance of 24 pS, an open probability of 0.816+/−0.067, an open time of 867+/−114 ms (mean +/− s.e.m., four patches) and the selectivity sequence I(−)>Cl(−)(much greater than) isethionate>gluconate. The I(−)/Cl(−)>>isethionate>gluconate. The I(−)Cl(−) permeability ratio was 1.48, corresponding to Eisenman sequence I. The type I Cl(−) channel was blocked by 2,2′-iminodibenzoic acid (DPC) and niflumic acid (2-[3-(trifluoromethyl)anilo]nicotinic acid). The removal of Ca(2+) from the Ringer's solution on the cytoplasmic side had no effect on channel activity. The second Cl(−) channel, type II, had a conductance of 8 pS, an open probability of 0.066+/−0.021 and an open time of 7.53+/−1.46 ms (mean +/− s.e.m., four patches). The high density and halide selectivity sequence of the type I Cl(−) channel is consistent with a role in transepithelial Cl(−) secretion under control conditions, but it remains to be determined whether these Cl(−) channels also mediate transepithelial Cl(−) secretion under diuretic conditions in the presence of leucokinin.

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