Secretion by the Malpighian Tubules of Rhodnius. The Movements of Ions and Water

1. The Malpighian tubules of Rhodnius will secrete at a normal rate in solutions containing no potassium ions and the rate is unaffected by changes in the potassium concentration, when the balance of cationic concentration is made up by sodium ions. 2. In the absence of sodium ions the rate of secretion is much reduced. The addition of very small amounts of sodium-containing solution brings about an abrupt recovery in the rate and thereafter the rate is unaffected by further increases in the sodium concentration. 3. In solutions containing either sodium or potassium with choline making up the balance of monovalent cations, the rate of fluid secretion depends linearly on the concentration of either sodium or potassium. 4. The tubules will concentrate either sodium or potassium when they are present at low concentration in the bathing fluid, even in the face of a very much larger concentration of the other cation. This suggests that there are separate mechanisms for the handling of these two ions. 5. Secretion can be supported by a solution containing ammonium ions in place of sodium and potassium. The tubules behave, at least in the short term, as if they were unable to distinguish ammonium from potassium. 6. Chloride ions appear to play a special role in that only bromide ions and, to a limited extent, nitrate ions will substitute for them. The rate of secretion depends linearly on the chloride concentration. 7. The tubules secrete a fluid which is practically iso-osmotic to the bathing fluid. The rate of secretion depends inversely on the osmolarity of the bathing fluid. The rate of movement of solute is little affected by these changes of osmolarity. It appears that water movements follow, and are closely linked with, solute movements. 8. Copper, cyanide, iodoacetate and azide ions and 2,4-dinitrophenol all stop secretion when added to the bathing medium. Ouabain, acetazolamide and mammalian ADH all have no effect on the rate of secretion and ouabain has no effect on the composition of the secreted fluid. 5-Hydroxytryptamine (serotonin) will stimulate a rapid flow of secretion. 9. Apparent neurosecretory axons have been found which supply the tubules. They do not contain enough diuretic activity to do more than play a minor role in diuresis. They may contribute to the rapid onset of diuresis or may affect the Malpighian tubules in some other way. 10. The evidence suggests that the tubules function by secreting chloride, potassium and sodium ions into the lumen (it is speculated that this may conceivably involve the active transport of all three ions) and that water movements closely follow these ion movements so that a rapid flow of iso-osmotic fluid is achieved.

Download Full-text

Transepithelial and intracellular potentials in isolated Malpighian tubules of tenebrionid beetle

AJP Renal Physiology ◽

10.1152/ajprenal.1987.252.4.f645 ◽

1987 ◽

Vol 252 (4) ◽

pp. F645-F653 ◽

Cited By ~ 1

Author(s):

S. W. Nicolson ◽

L. C. Isaacson

Keyword(s):

Membrane Potential ◽

Basal Membrane ◽

Fluid Secretion ◽

Malpighian Tubules ◽

Bathing Medium ◽

Intracellular Potentials ◽

Tenebrionid Beetle ◽

Fold Reduction ◽

K Concentration ◽

Opposing Effects

Malpighian tubules of Onymacris plana (Coleoptera: Tenebrionidae) have been isolated for measurement of transepithelial and intracellular potentials, before and during stimulation of fluid secretion. In a bathing medium resembling the hemolymph composition of the insect, the transepithelial potential (VT) was approximately 13 mV, lumen positive. VT was subject to drift and frequently showed super-imposed regular oscillations, which were apparently action potentials associated with contractions of muscle fibers running along the tubules. Although tubules of Onymacris are approximately 8 cm long, the basal membrane potential (Vb) did not vary with distance along the tubule, averaging -31 mV. Addition of adenosine 3',5'-cyclic monophosphate (cAMP) or diuretic hormone (DH) homogenate to the bathing medium had no effect on Vb, but opposing effects on VT: cAMP caused it to increase to 60 mV, whereas DH homogenate caused a rapid drop in VT to almost zero. Ion substitutions in the bathing medium showed that under control conditions beetle tubules possessed appreciable basal permeability to both K and Cl ions, with a 10-fold reduction in bath K concentration hyperpolarizing Vb by 54 mV. The basal K and Cl channels were partially blocked by barium and thiocyanate ions, respectively. Stimulation with cAMP increased the apical membrane potential (Va) and significantly reduced the Cl permeability of the basal membrane, whereas its Na permeability remained negligible.

Download Full-text

Secretion by the Malpighian tubules of Rhodnius prolixus stal: electrical events

Journal of Experimental Biology ◽

10.1242/jeb.110.1.275 ◽

1984 ◽

Vol 110 (1) ◽

pp. 275-290 ◽

Cited By ~ 11

Author(s):

M. J. O'Donnell ◽

S. H. Maddrell

Keyword(s):

Apical Membrane ◽

Electrical Response ◽

Basal Membrane ◽

Fluid Secretion ◽

Malpighian Tubules ◽

Chloride Permeability ◽

Bathing Medium ◽

Before And After ◽

Electrical Gradient ◽

Cation Pump

Transepithelial and intracellular potentials have been simultaneously recorded from Rhodnius upper Malpighian tubules before and after stimulation of fluid secretion. The transepithelial electrical response to the diuretic hormone mimic 5-hydroxytryptamine (5-HT) was triphasic; recordings of intracellular potential changes indicated that the three phases represented successive events at the apical membrane. Depolarizations produced by increasing the bathing medium potassium concentration indicated that the basal membrane was much more permeable to potassium than to sodium. Electrical responses to chloride-free saline were inconsistent with a significant basal membrane chloride permeability. Chloride movements across the basal membrane were opposed by an electrical gradient of about 65 mV. The results of experiments in which tubules were exposed to chloride-free saline or sodium-free saline suggested that chloride entry into the cells was linked to the entry of Na+ and K+. The effects of furosemide and bumetanide upon secretion and potential changes suggested that chloride crossed the basal membrane through co-transport with Na+ and K+. Chloride probably crosses the apical membrane into the lumen passively in response to a favourable electrical gradient of about 35 mV. Cations must be actively pumped into the lumen against an electrical gradient of 35 mV. Our results support previous evidence for an apical cation pump which actively transports Na and K into the lumen. A tentative model of ionic movements during fluid secretion is presented. It is suggested that the apical cation pump maintains sodium at low intracellular concentrations, thereby maintaining a favourable gradient for entry of Na+ through the proposed basal co-transport step. The suggested stoichiometry is Na+:K+:2 Cl-.

Download Full-text

A Critical Study of the Evidence for Peripheral Inhibitory Axons in Insects

Journal of Experimental Biology ◽

10.1242/jeb.49.1.201 ◽

1968 ◽

Vol 49 (1) ◽

pp. 201-222

Author(s):

P. N. R. USHERWOOD

Keyword(s):

Potassium Concentration ◽

Chloride Concentration ◽

Chloride Ions ◽

Resting Potential ◽

Equilibrium Potential ◽

Critical Study ◽

Muscle Fibres ◽

High Concentration ◽

Bathing Medium

1. The metathoracic anterior coxal adductor (a.c.a.) muscle of the locust and the grasshopper is innervated by a peripheral inhibitory axon similar to the inhibitory axon which innervates the metathoracic extensor tibiae muscles of these insects. No evidence was found to justify calling this axon an inhibitory-conditioning axon. 2. Hyperpolarizing inhibitory postsynaptic potentials (IPSPs) are normally recorded from a.c.a. muscle fibres during stimulation of this axon, and if the bathing medium contains a high concentration of potassium ions the tonic fibres of the a.c.a. muscle relax slightly during inhibitory stimulation. 3. The IPSPs are chloride potentials and can be converted to depolarizing responses by changing either the external or internal chloride concentration of the a.c.a. muscle fibres. Depolarizing IPSPs are frequently accompanied by small contractions of a.c.a. muscle fibres innervated by the inhibitory axon. 4. The a.c.a. muscle fibres are permeable to potassium and chloride ions but influx of potassium chloride is much faster than efflux. Therefore when a.c.a. muscle fibres are loaded with chloride by exposing them to high-K saline (20-100 m-equiv. potassium/l.) and are then returned to normal (10 m-equiv. potassium/l.) saline the internal chloride concentration remains elevated for some time and during this period the equilibrium potential for the inhibitory response is less negative than the resting potential and the IPSPs are depolarizing. 5. Depolarizing IPSPs are usually recorded from a.c.a. muscle fibres of locusts and grasshoppers when these fibres are transferred from their normal bathing medium, haemolymph, to 10 K saline. Probably the main reason for this reversal of the IPSPs is the entry of KCl into the muscle fibres during dissection of the nerve-muscle preparations. Large quantities of KCl would be released into the environment surrounding these preparations from muscle fibres cut and removed during dissection. 6.Depolarizing IPSPs were more frequently recorded from muscle fibres of grassfed locusts than from fibres of starved locusts. The potassium concentration of haemolymph of grass fed locusts is higher than that of locust saline (10 m-equiv./l.). 7. The potassium concentration of locust haemolymph presumably fluctuates in vivo but these fluctuations are too slow to affect the sign of the IPSP. The IPSPs are therefore always hyperpolarizing in vivo. 8. The effect of changes in the potassium concentration of the bathing medium on the magnitude and polarity of the IPSP could account for the diverse responses recorded previously from a.c.a. muscle fibres of locusts and grasshoppers.

Download Full-text

Fluid secretion by the malpighian tubules of the tsetse fly Glossina morsitans: the effects of ouabain, ethacrynic acid and amiloride

Journal of Experimental Biology ◽

10.1242/jeb.65.2.323 ◽

1976 ◽

Vol 65 (2) ◽

pp. 323-332

Author(s):

J. D. Gee

Keyword(s):

Sodium Transport ◽

Cardiac Glycoside ◽

Ethacrynic Acid ◽

Fluid Secretion ◽

Sodium Concentration ◽

Tsetse Fly ◽

Malpighian Tubules ◽

Sodium Ions ◽

Glossina Morsitans ◽

Sodium And Potassium

The effects of three inhibitors of sodium transport on the secretion of fluid by the Malpighian tubules of Glossina morsitans have been observed. The cardiac glycoside, ouabain, affects neither the rate of secretion nor the sodium concentration of the fluid secreted when isolated tubules are bathed by solutions containing a range of sodium and potassium concentrations. Secretion is inhibited, however, by ethacrynic acid and amiloride. The results confirm that fluid secretion by the Malpighian tubules of this insect is dependent on the active transport of sodium ions and show that Na+/k+ exchange pumps are not involved in this process.

Download Full-text

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

Journal of Experimental Biology ◽

10.1242/jeb.137.1.265 ◽

1988 ◽

Vol 137 (1) ◽

pp. 265-276 ◽

Cited By ~ 8

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.

Download Full-text

Mechanisms of K+ uptake across the basal membrane of malpighian tubules of Formica polyctena: the effect of ions and inhibitors.

Journal of Experimental Biology ◽

10.1242/jeb.195.1.123 ◽

1994 ◽

Vol 195 (1) ◽

pp. 123-145 ◽

Cited By ~ 11

Author(s):

A Leyssens ◽

S Dijkstra ◽

E Van Kerkhove ◽

P Steels

Keyword(s):

Membrane Potential ◽

Electrical Potential ◽

Basal Membrane ◽

Fluid Secretion ◽

Malpighian Tubules ◽

K Uptake ◽

Bathing Medium ◽

K Channels ◽

High K ◽

K Concentration

In the presence of 6 mmol l-1 Ba2+, known to block the K+ channels in the basal membrane, a rise in bath [K+] ([K+]bl) induced an increase in intracellular K+ concentration ([K+]i) similar in amount and in time course to that obtained in the absence of Ba2+. The presence of active and passive (other than through K+ channels) K+ uptake mechanisms across the basal membrane was investigated in different bath K+ concentrations. Dihydro-ouabain (10(-3) mol l-1), a blocker of the Na+/K(+)-ATPase, tested in low bath [K+], and Sch28080 (10(-4) mol l-1), a K+/H(+)-ATPase inhibitor, were without effect on fluid secretion. Dihydro-ouabain was also without effect on electrical potential differences either in the absence or in the presence of Ba2+. Vanadate (10(-3) mol l-1), in contrast, strongly reduced fluid secretion not only in control solution but also in high-K+, Na(+)-free medium and reduced the transepithelial and the apical membrane potential differences but not the basal membrane potential difference of [K+]i. Omitting Na+ from the bathing medium, replacing Cl- by Br- or applying bumetanide (10(-5) mol l-1) inhibited fluid secretion only in a low-K+ (10 mmol l-1) medium. In 51 mmol l-1 [K+]bl, omitting Na+ was without effect and 10(-4) mol l-1 bumetanide was needed to inhibit secretion. Replacing Cl- by Br- stimulated fluid secretion at this K+ concentration. Bumetanide (10(-4) mol l-1) had no effect in 113 mmol l-1 [K+]bl. Bumetanide (10(-4) mol l-1) in 51 mmol l-1 [K+]bl did not affect membrane potentials, did not lower [K+]i and did not affect the rise in [K+]i observed on an increase in [K+]bl. The results were summarized in a model proposing that K+ channels play a dominant role in high-K+ (113 mmol l-1) bathing medium. A K+/Cl- cotransporter may become more important in 51 mmol l-1 [K+]bl and a K+/Na+/2Cl- cotransporter may gain in importance in 10 mmol l-1 [K+]bl. Active mechanisms for K+ uptake across the basal membrane seem to play no detectable role in sustaining fluid secretion. The response to vanadate might be due to an effect on the apical electrogenic H+ pump.

Download Full-text

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

Journal of Experimental Biology ◽

10.1242/jeb.202.11.1561 ◽

1999 ◽

Vol 202 (11) ◽

pp. 1561-1570 ◽

Cited By ~ 20

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.

Download Full-text

Active transport of sodium by the malpighian tubules of the tsetse fly Glossian morsitans

Journal of Experimental Biology ◽

10.1242/jeb.64.2.357 ◽

1976 ◽

Vol 64 (2) ◽

pp. 357-368

Author(s):

J. D. Gee

Keyword(s):

Cell Function ◽

Malpighian Tubule ◽

Active Role ◽

Fluid Secretion ◽

Sodium Concentration ◽

Tsetse Fly ◽

Malpighian Tubules ◽

Bathing Solution ◽

Bathing Medium

Isolated Malpighian tubules of Glossina morsitans are able to transport sodium against its concentration gradient. Their rate of secretion is dependent on the sodium concentration of the bathing medium. Potassium must be present in the bathing solution for rapid secretion to be maintained, but it does not play an active role in fluid secretion. Lithium and ammonium ions are able to substitute partially for sodium, other monovalent cations cannot. Ouabain does not affect rapid secretion by Glossina tubules in vitro. Conclusions drawn from the results are incorporated into a model of Malpighian tubule cell function in this insect.

Download Full-text

Dibutyryl cAMP activates bumetanide-sensitive electrolyte transport in Malpighian tubules

AJP Cell Physiology ◽

10.1152/ajpcell.1991.261.3.c521 ◽

1991 ◽

Vol 261 (3) ◽

pp. C521-C529 ◽

Cited By ~ 66

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.

Download Full-text