NHE8 mediates amiloride-sensitive Na+/H+exchange across mosquito Malpighian tubules and catalyzes Na+and K+transport in reconstituted proteoliposomes

2007 ◽  
Vol 292 (5) ◽  
pp. F1501-F1512 ◽  
Author(s):  
Wanyoike Kang'ethe ◽  
Karlygash G. Aimanova ◽  
Ashok K. Pullikuth ◽  
Sarjeet S. Gill
Keyword(s):  
Fluid Transport ◽  
Ion Homeostasis ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Yeast Cells ◽  
Kinetic Properties ◽  
Sodium Load ◽  
Electrophysiological Studies ◽  
Reconstituted Membranes ◽  
Apical Membranes

Following a blood meal, the mosquito Aedes aegypti will have acquired an enormous sodium load that must be rapidly excreted to restore ion homeostasis. It is a process that demands robust sodium and fluid transport capabilities. Even though the identities of the components involved in this ion transport across the mosquito Malpighian tubule epithelia have not been completely determined, electrophysiological studies suggest the contribution of a Na+/H+exchanger extruding cations into the lumen driven secondarily by the proton gradient created by the V-type H+-ATPase in the tubules' apical membrane. We have identified the putative exchanger and designated it AeNHE8. Immunolocalization studies demonstrated that AeNHE8 is expressed in the apical membranes of Malpighian tubules, gastric caecae, and rectum. When heterologously expressed in salt-sensitive yeast cells lacking Na+extrusion and Na+/H+exchange proteins, AeNHE8 rescues the salt-sensitive phenotype and restores the cells' ability to grow in high NaCl media. Furthermore, heterologous expression of AeNHE8 in NHE-deficient fibroblast cells results in an amiloride-sensitive22Na+uptake. To determine the exchanger's kinetic properties, we reconstituted membranes from yeast cells expressing the protein into lipid proteoliposomes and assayed for cation-dependent H+exchange by fluorimetric methods. Our results indicate that AeNHE8 mediates saturable exchange of Na+and K+for H+. We propose that AeNHE8 may be coupled to the inward H+gradient across the Malpighian tubules and plays a role in the extrusion of excess sodium and potassium while maintaining steady intracellular pH in the principal cells.

scholarly journals Functional Plasticity of the gut and the Malpighian tubules underlies cold acclimation and mitigates cold-induced hyperkalemia in Drosophila melanogaster

2017 ◽  
Author(s):  
Gil Y. Yerushalmi ◽  
Lidiya Misyura ◽  
Heath A. MacMillan ◽  
Andrew Donini
Keyword(s):  
Water Balance ◽  
Cold Acclimation ◽  
Low Temperatures ◽  
Fluid Transport ◽  
Epithelial Transport ◽  
Ion Homeostasis ◽  
Malpighian Tubule ◽  
Adult Emergence ◽  
Fluid Secretion ◽  
Malpighian Tubules

AbstractAt low temperatures, Drosophila, like most insects, lose the ability to regulate ion and water balance across the gut epithelia, which can lead to a lethal increase of [K+] in the hemolymph (hyperkalemia). Cold-acclimation, the physiological response to low temperature exposure, can mitigate or entirely prevent these ion imbalances, but the physiological mechanisms that facilitate this process are not well understood. Here, we test whether plasticity in the ionoregulatory physiology of the gut and Malpighian tubules of Drosophila may aid in preserving ion homeostasis in the cold. Upon adult emergence, D. melanogaster females were subjected to seven days at warm (25°C) or cold (10°C) acclimation conditions. The cold acclimated flies had a lower critical thermal minimum (CTmin), recovered from chill coma more quickly, and better maintained hemolymph K+ balance in the cold. The improvements in chill tolerance coincided with increased Malpighian tubule fluid secretion and better maintenance of K+ secretion rates in the cold, as well as reduced rectal K+ reabsorption in cold-acclimated flies. To test whether modulation of ion-motive ATPases, the main drivers of epithelial transport in the alimentary canal, mediate these changes, we measured the activities of Na+-K+-ATPase and V-type H+-ATPase at the Malpighian tubules, midgut, and hindgut. Na+/K+-ATPase and V-type H+-ATPase activities were lower in the midgut and the Malpighian tubules of cold-acclimated flies, but unchanged in the hindgut of cold acclimated flies, and were not predictive of the observed alterations in K+ transport. Our results suggest that modification of Malpighian tubule and gut ion and water transport likely prevents cold-induced hyperkalemia in cold-acclimated flies and that this process is not directly related to the activities of the main drivers of ion transport in these organs, Na+/K+- and V-type H+-ATPases.Summary StatementAt low temperatures, insects lose the ability to regulate ion and water balance and can experience a lethal increase in hemolymph [K+]. Previous exposure to low temperatures can mitigate this effect and improve chill tolerance. Here, we show that plasticity of ion and fluid transport across the Malpighian tubule and rectal epithelia likely drive this response.

Unique electrophysiological effects of dinitrophenol in Malpighian tubules

1992 ◽  
Vol 263 (3) ◽  
pp. R609-R614 ◽  
Author(s):  
T. L. Pannabecker ◽  
D. J. Aneshansley ◽  
K. W. Beyenbach
Keyword(s):  
Apical Membrane ◽  
Basolateral Membrane ◽  
Membrane Resistance ◽  
Malpighian Tubules ◽  
Membrane Voltage ◽  
Shunt Resistance ◽  
Electrophysiological Studies ◽  
Apical Membranes ◽  
Conductive Properties ◽  
Electrophysiological Effects

In the course of electrophysiological studies of Malpighian tubules of the mosquito Aedes aegypti, we have found unusual effects of 2,4-dinitrophenol (DNP) that offer new insights into the electrogenic and conductive properties of the tubule. DNP (10(-4)M) depolarized the basolateral membrane voltage from -58.0 to -3.3 mV, and it depolarized the apical membrane voltage from 110.6 to 8.9 mV. In parallel the transepithelial electrical resistance increased from 11.4 to 16.8 k omega.cm, and the fractional resistance of the apical membrane increased from 0.32 to 0.57. On the assumption that measures of transepithelial resistance in the presence of DNP approach the shunt resistance, the experimental results indicate the following characteristics for the equivalent circuit of the tubule: 1) a shunt resistance that is approximately one-half the transcellular resistance, 2) low and high electromotive forces, respectively, at the basolateral and apical membranes of principal cells, 3) an electrogenic pump at the apical membrane, and 4) a basolateral membrane voltage that is due mostly to the voltage developed by current flow across the basolateral membrane resistance.

scholarly journals Physiology, Development, and Disease Modeling in the Drosophila Excretory System

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
Keyword(s):  
Human Disease ◽  
Fluid Transport ◽  
Disease Modeling ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Excretory System ◽  
Anatomy And Physiology ◽  
Disease Biology ◽  
Organ Systems ◽  
And Function

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.

Stimulation of sodium transport and fluid secretion by ouabain in an insect malpighian tubule

1988 ◽  
Vol 137 (1) ◽  
pp. 265-276 ◽  
Author(s):  
S. H. Maddrell ◽  
J. A. Overton
Keyword(s):  
Fluid Flow ◽  
Sodium Transport ◽  
Fluid Transport ◽  
Electrical Potential ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Malpighian Tubules ◽  
Sodium Ions ◽  
Tubule Cells ◽  
Stimulation Of

Ouabain, at all concentrations higher than 2 × 10(−7) mol l-1, stimulates the rate at which the Malpighian tubules of the insect, Rhodnius, transport sodium ions and fluid into the lumen. An effect on paracellular movement of sodium ions is unlikely because ouabain makes the electrical potential of the lumen more positive, which would slow diffusion of sodium into the lumen. Radioactive ouabain binds to the haemolymph-facing sides of the tubule cells but not to the luminal face. This binding is reduced in the presence of elevated levels of potassium or of non-radioactive ouabain. Bound ouabain is only slowly released on washing in ouabain-free saline. The evidence suggests that there is a Na+/K+-ATPase on the outer (serosal) membranes of the tubules. Such a pump would transport sodium in a direction opposed to the flow of ions and water involved in fluid transport; poisoning it with ouabain would remove this brake, and fluid flow and sodium transport would increase, as observed.

Fine Structure of the Malpighian Tubules of the Mosquito, Culex pipiens

1971 ◽  
Vol 29 ◽  
pp. 402-403
Author(s):  
Brendan Clifford
Keyword(s):  
Fine Structure ◽  
Culex Pipiens ◽  
Stellate Cell ◽  
Malpighian Tubule ◽  
Cell Types ◽  
Instar Larva ◽  
Malpighian Tubules ◽  
Calliphora Erythrocephala ◽  
Distinct Cell ◽  
Basal Plasma

An ultrastructural investigation of the Malpighian tubules of the fourth instar larva of Culex pipiens was undertaken as part of a continuing study of the fine structure of transport epithelia.Each of the five Malpighian tubules was found to be morphologically identical and regionally undifferentiated. Two distinct cell types, the primary and stellate, were found intermingled along the length of each tubule. The ultrastructure of the stellate cell was previously described in the Malpighian tubule of the blowfly, Calliphora erythrocephala by Berridge and Oschman.The basal plasma membrane of the primary cell is extremely irregular, giving rise to a complex interconnecting network of basal channels. The compartments of cytoplasm entrapped within this system of basal infoldings contain mitochondria, free ribosomes, and small amounts of rough endoplasmic reticulum. The mitochondria are distinctive in that the cristae run parallel to the long axis of the organelle.

A Study of the Malpighian Tubules of the Pill Millipede, Glomeris marginata (Villers)

1974 ◽  
Vol 60 (1) ◽  
pp. 41-51
Author(s):  
PATRICIA ANNE FARQUHARSON
Keyword(s):  
Size Distribution ◽  
Fluid Transport ◽  
Molecular Size ◽  
Malpighian Tubules ◽  
Inverse Relation ◽  
Fluid Production ◽  
Molecular Size Distribution ◽  
Molecular Sieving ◽  
Tubule Fluid ◽  
Tubule Wall

1. Tubule fluid:medium ratios (TF/M) have been measured for inulin, glucose, LMWD and HMWD. These TF/M ratios were surprisingly high. 2. The tubule appears to act as a molecular filter; that is to say, molecules move through the tubule wall in inverse relation to their size. This is best illustrated using polyvinyl pyrrolidone as a tracer. The molecular size distribution of PVP fractions present in tubule fluid differs markedly from the molecular size distribution of PVP in the bathing Ringer. 3. No correlation can be made between the inulin and glucose TF/M and the rate of fluid production. However, the inverse relationship between TF/M and rate of fluid production for dextrans indicates a molecular sieving effect. 4. The significance of these results is discussed with reference to models of fluid transport.

THE EFFECTS OF MANDUCA SEXTA DIURETIC HORMONE ON FLUID TRANSPORT BY THE MALPIGHIAN TUBULES AND CRYPTONEPHRIC COMPLEX OF MANDUCA SEXTA

1993 ◽  
Vol 178 (1) ◽  
pp. 231-243 ◽  
Author(s):  
N. Audsley ◽  
G. M. Coast ◽  
D. A. Schooley
Keyword(s):  
Cyclic Amp ◽  
Manduca Sexta ◽  
Fluid Transport ◽  
Basal Membrane ◽  
Fluid Secretion ◽  
Hormonal Control ◽  
Malpighian Tubules ◽  
Proximal Tubules ◽  
Fluid Absorption ◽  
Diuretic Hormone

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.

Anti-diuresis in the blood-feeding insect Rhodnius prolixus Stål: the peptide CAP2b and cyclic GMP inhibit Malpighian tubule fluid secretion.

1997 ◽  
Vol 200 (17) ◽  
pp. 2363-2367 ◽  
Author(s):  
M C Quinlan ◽  
N J Tublitz ◽  
M J O'Donnell
Keyword(s):  
Cyclic Gmp ◽  
Physiological Role ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Blood Feeding ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Tubule Fluid ◽  
Secretion Rates

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.

The permeability properties of septate junctions in Malpighian tubules of Rhodnius

1987 ◽  
Vol 88 (2) ◽  
pp. 251-265 ◽  
Author(s):  
H.B. Skaer ◽  
S.H. Maddrell ◽  
J.B. Harrison
Keyword(s):  
Structural Characteristics ◽  
Malpighian Tubule ◽  
Malpighian Tubules ◽  
Septate Junctions ◽  
Blood Sucking ◽  
Effects Of Ph ◽  
Intercellular Clefts ◽  
Luminal Flow ◽  
Tubule Cells ◽  
Positively Charged

This paper describes the structural characteristics and permeability properties of the smooth septate junctions between the upper Malpighian tubule cells of a blood-sucking bug, Rhodnius prolixus. The permeability of the paracellular route was tested only for solutes that could be demonstrated not to cross the epithelium via the cellular route. The intercellular clefts were readily permeated by sucrose, inulin and polyethylene glycol (PEG), showing a higher permeability to molecules of smaller radius (PEG versus sucrose). Negatively charged molecules permeated the clefts more readily than positively charged ones. The effects of pH, urea and luminal flow rate on permeability were studied. The results are discussed in relation to the physiological tightness of the Malpighian tubules to certain solutes and to its function as an excretory epithelium.

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.

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