Passage of solutes through walls of Malpighian tubules of Rhodnius by paracellular and transcellular routes

1984 ◽  
Vol 246 (5) ◽  
pp. R759-R769 ◽  
Author(s):  
M. J. O'Donnell ◽  
S. H. Maddrell ◽  
B. O. Gardiner
Keyword(s):  
Cell Membrane ◽  
Time Course ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Bathing Medium ◽  
Tubule Cells

The walls of isolated upper Malpighian tubules of the insect, Rhodnius prolixus, are much more permeable to small relatively unchanged solutes (ethanol, xylose, and mannitol) than to larger or more charged solutes (acetate, glycine, tyrosine, and inulin). The more permeable solutes rapidly reach concentrations in the tubule cells equivalent to their concentrations in the bathing medium; the less permeable solutes do not penetrate into the cells. The time course of accumulation of permeable solute in the cells matches the time course of the appearance of solute in the lumen. Substances injected into the hemolymph of fed R. prolixus appear in the urine at concentrations predictable from the permeability of in vitro tubules, supporting the idea that the in vitro permeability of the tubules is representative of their properties in the intact insect. It is suggested that the rapid transcellular penetration of small solutes through the Malpighian tubules reflects the large areas of cell membrane. The area of cell membrane exceeds that of the paracellular clefts by a factor of 10(5).

scholarly journals Action of activated 27,000 Mr toxin from Bacillus thuringiensis var. israelensis on Malpighian tubules of the insect, Rhodnius prolixus

1989 ◽  
Vol 94 (3) ◽  
pp. 601-608
Author(s):  
S.H. Maddrell ◽  
J.A. Overton ◽  
D.J. Ellar ◽  
B.H. Knowles
Keyword(s):  
Bacillus Thuringiensis ◽  
Cell Membrane ◽  
Basal Cell ◽  
Time Course ◽  
Fluid Secretion ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Bathing Solution ◽  
Concentrated Solutions ◽  
Intracellular Milieu

The action of activated 27,000 Mr toxin from Bacillus thuringiensis var. israelensis (Bti toxin) on Malpighian tubules of Rhodnius prolixus has been investigated. Its binding to the tubules is slowed by low temperature but is not prevented even at 0 degree C. The binding is less effective at pH 10 than at pH7. Pretreatment of the tubules with 0.1 mmol l-1 ouabain or bumetanide or 1 mumol l-1 5-hydroxytryptamine did not affect the toxicity of the toxin. The toxin causes very large changes in the trans-epithelial potential difference; it changes from 40 mV, lumen negative, often to more than 100 mV, lumen positive. This reflects an initial collapse of the potential of the basal cell membrane, followed by a large positive-going potential change at the luminal cell membrane. Just prior to the effects of the toxin on rapid fluid secretion, the basal cell membrane becomes permeable to sucrose molecules. Raffinose at 170 mmol l-1 in the bathing solution does not protect the tubules from Bti toxin action but dextran, Mr5000, at 60 mmol l-1 significantly delayed failure of fluid secretion and, even more, the onset of staining of the tubule cells with Trypan Blue. Exposing tubules to saline that is calcium-free and/or magnesium-free, or has a composition adjusted to be similar to that of the intracellular milieu, does not affect the time course of failure of fluid secretion induced by the toxin. There is no evidence that effective aggregates of Bti toxin molecules are formed in concentrated solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracellular ion activities in Malpighian tubule cells ofRhodnius prolixus: evaluation of Na+-K+-2Cl-cotransport across the basolateral membrane

2002 ◽  
Vol 205 (11) ◽  
pp. 1645-1655 ◽  
Author(s):  
Juan P. Ianowski ◽  
Robert J. Christensen ◽  
Michael J. O'Donnell
Keyword(s):  
Basolateral Membrane ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Rhodnius Prolixus ◽  
Passive Movement ◽  
Malpighian Tubules ◽  
Intracellular Ion Activities ◽  
Ion Activities ◽  
Tubule Cells ◽  
Electrochemical Potentials

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.

The fate of calcium in the diet of Rhodnius prolixus: storage in concretion bodies in the Malpighian tubules

1991 ◽  
Vol 157 (1) ◽  
pp. 483-502 ◽  
Author(s):  
S. H. Maddrell ◽  
G. Whittembury ◽  
R. L. Mooney ◽  
J. B. Harrison ◽  
J. A. Overton ◽  
...  
Keyword(s):  
Calcium Concentration ◽  
Rhodnius Prolixus ◽  
The Body ◽  
Malpighian Tubules ◽  
Calcium Deposition ◽  
X Ray Diffraction ◽  
Calcium Deposits ◽  
Membrane Bound ◽  
High Concentrations

We have investigated the fate of the large amounts of calcium ingested by Rhodnius prolixus in its meals of blood. 45Ca2+ injected into the haemolymph or fed to fifth-stage Rhodnius reared on rabbits is accumulated at high concentrations in the cells of the upper Malpighian tubules; very little is excreted from the body This 45Ca2+ accumulation goes on continuously for at least 12 days and the rate of uptake is increased several-fold within 3–4 days of a meal. The extent of calcium accumulation in tubule cells is correlated with the presence of intracellular membrane-bound concretion bodies, which are therefore likely sites of calcium deposition. X-ray diffraction showed that the calcium deposits are non-crystalline. Tubules from rabbit-fed fifth-stage Rhodnius contain 410 mmol l-1 calcium; in those from chicken-fed insects the calcium concentration is over 1 mol l-1; and in those fed in vitro on heparinised low-K+ sheep blood the calcium concentration is only 21 mmol l-1. The concentration of calcium in the haemolymph in all these insects was 8 mmol l-1 and its activity determined by an ion-selective electrode was 2.5 mmol l-1. 45Ca2+ deposited in the tubules is readily exchangeable, but the efflux preferentially passes to the haemolymph side of the tubule epithelium. The ability to sequester calcium in the Malpighian tubules may prevent calcium from interfering with reabsorptive processes in the rectum.

scholarly journals Studies on the time course and rate-limiting steps in the activation of adenylate cyclase in rat liver by cholera toxin

1978 ◽  
Vol 173 (1) ◽  
pp. 59-64 ◽  
Author(s):  
J Fischer ◽  
T R Kohler ◽  
L G Lipson ◽  
J Flores ◽  
P A Witkum ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Adenylate Cyclase ◽  
Cholera Toxin ◽  
Rat Liver ◽  
Time Course ◽  
Intact Cell ◽  
Intact Cells ◽  
Maximal Stimulation ◽  
Rate Limiting

Cholera toxin stimulates adenylate cyclase in rat liver after intravenous injection. The stimulation follows a short latent period of 10min, and maximum stimulation was attained at 120min. Half-maximal stimulation was achieved at 35min. In contrast with this lengthy time course in the intact cell, adenylate cyclase in broken-cell preparations of rat liver in vitro were maximally stimulated by cholera toxin (in the presence of NAD+) in 20min with half-maximal stimulation in 8min. Binding of cholera toxin to cell membranes by the B subunits is followed by translocation of the A subunit into the cell or cell membrane, and separation of the A1 polypeptide chain from the A2 chain by disulphide-bond reduction, and finally activation of adenylate cyclase by the A1 chain and NAD+. As the binding of cholera toxin is rapid, two possible rate-limiting steps could be the determinants of the long time course of action. These are translocation of the A1 chain from the outside of the cell membrane to its site of action (this includes the time required for separation from the whole toxin) or the availability of NAD+ for activation. When NAD+ concentrations in rat liver were elevated 4-fold, by the administration of nicotinamide, no change in the rate of activation of adenylate cyclase by cholera toxin was observed. Thus the intracellular concentration of NAD+ is not rate-limiting and the major rate-limiting determinant in intact cells must be between the time of toxin binding to the cell membrane and the appearance of subunit A1 at the enzyme site.

In vitro effect of Canavalia ensiformis urease and the derived peptide Jaburetox-2Ec on Rhodnius prolixus Malpighian tubules

2009 ◽  
Vol 55 (3) ◽  
pp. 255-263 ◽  
Author(s):  
Fernanda Stanisçuaski ◽  
Victoria Te Brugge ◽  
Célia R. Carlini ◽  
Ian Orchard
Keyword(s):  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Canavalia Ensiformis ◽  
Vitro Effect

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

1974 ◽  
Vol 60 (1) ◽  
pp. 13-28
Author(s):  
PATRICIA ANNE FARQUHARSON
Keyword(s):  
Resistance Mechanism ◽  
Ringer Solution ◽  
Malpighian Tubules ◽  
Metabolic Inhibitors ◽  
Bathing Medium ◽  
Diuretic Hormone ◽  
Fluid Production ◽  
Tubule Fluid ◽  
Secretion Rates

1. The Malphigian tubules of the pill millipede, Glomeris marginata are described. 2. After the necessary analyses of the haemolymph had been carried out a Ringer solution was prepared which would support the secretion of tubule fluid in vitro. 3. The reasons for the variation and deterioration in the secretion rates of control tubules are discussed. The presence of various amino acids in the bathing medium appears to be essential for fluid production. 4. There would not appear to be a diuretic hormone involved in the control of secretion; and 5-HT, cylic AMP and theophylline will not stimulate fluid production. 5. Secretion is inhibited by the metabolic inhibitors 2,4-DNP, azide and cyanide. The relative insensitivity shown to the latter two compounds is probably associated with a resistance mechanism.

Analysis of epithelial transport by measurement of K+, Cl- and pH gradients in extracellular unstirred layers: ion secretion and reabsorption by Malpighian tubules of Rhodnius prolixus

1997 ◽  
Vol 200 (11) ◽  
pp. 1627-1638 ◽  
Author(s):  
KA Collier ◽  
MJ O'Donnell
Keyword(s):  
Epithelial Transport ◽  
Malpighian Tubule ◽  
Fluid Secretion ◽  
Rhodnius Prolixus ◽  
Malpighian Tubules ◽  
Unstirred Layer ◽  
Ph Gradients ◽  
In Situ Preparation

Summary The pH and concentrations of K+ and Cl- in the unstirred layer (USL) associated with the basolateral surfaces of the upper and lower Malpighian tubules of Rhodnius prolixus were measured using extracellular ion-selective microelectrodes. When stimulated with 5-hydroxytryptamine (5-HT) in vitro, the upper Malpighian tubule secretes Na+, K+, Cl- and water at high rates; the lower Malpighian tubule reabsorbs K+ and Cl- but not water. Concentrations of K+ and Cl- in the unstirred layer of the lower Malpighian tubule ([K+]USL, [Cl-]USL) were greater than those in the bathing saline, consistent with the accumulation of K+ and Cl- in the USL during 5-HT-stimulated KCl reabsorption. [K+]USL exceeded [K+]Bath by as much as 5.3-fold. Calculations of K+ flux based on measurements of [K+]USL at various distances from the tubule surface agreed well with flux calculated from the rate of fluid secretion and the change in K+ concentration of the secreted fluid during passage through the lower tubule. Concentrations of K+ in the unstirred layer of the upper Malpighian tubule were reduced relative to those in the bathing saline, consistent with depletion of K+ from the USL during 5-HT-stimulated secretion of K+ from bath to lumen. Changes in [K+]USL during 5-HT-stimulated K+ secretion from single upper Malpighian tubule cells could be resolved. Although differences between [K+]USL and [K+]Bath were apparent for upper and lower tubules in an in situ preparation, they were reduced relative to the differences measured using isolated tubules. We suggest that convective mixing of the fluids around the tubules by contractions of the midgut and hindgut reduces, but does not eliminate, differences between [K+]USL and [K+]Bath in situ. The USL was slightly acidic relative to the bath in 5-HT-stimulated upper and lower tubules; contributions to USL acidification are discussed. The results also show that the techniques described in this paper can resolve rapid and localized changes in ion transport across different regions of Malpighian tubules in response to stimulants or inhibitors of specific membrane transporters.

scholarly journals Changes in cyclic AMP and cyclic GMP concentrations during the action of 5-hydroxytryptamine on an insect salivary gland

1980 ◽  
Vol 192 (1) ◽  
pp. 247-255 ◽  
Author(s):  
J P Heslop ◽  
M J Berridge
Keyword(s):  
Salivary Gland ◽  
Cyclic Amp ◽  
Cyclic Gmp ◽  
Time Course ◽  
Phosphodiesterase Inhibitor ◽  
Fluid Secretion ◽  
Fast Response ◽  
Bathing Medium ◽  
Calliphora Erythrocephala

Salivary glands from adult blowflies (Calliphora erythrocephala Meigen) were studied in vitro. The time course of changes in cyclic AMP content of the glands was followed at different concentration of 5-hydroxytryptamine. There was an immediate biphasic rise and fall in cyclic AMP content, following by a slower rise and subsequent gradual decline. The initial rise preceded the onset of fluid secretion by the glands. Rises in cyclic AMP content were inhibited by compound RMI 12330 A (an adenylate cyclase inhibitor) and were halted after about 15-20s if the glands were deprived of Ca2+. Theophylline (a phosphodiesterase inhibitor) abolished the decline phase of the fast response, Losses of cyclic AMP from the glands either to the bathing medium or to the saliva were small and could not account for the rapid fall found. Evidence is presented that cyclic GMP is not involved in the process of initiating secretion in the blowfly salivary gland.

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

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.

Comparison of High Purity Factor IX Concentrates and a Prothrombin Complex Concentrate in a Canine Model of Thrombogenicity

1991 ◽  
Vol 66 (05) ◽  
pp. 609-613 ◽  
Author(s):  
I R MacGregor ◽  
J M Ferguson ◽  
L F McLaughlin ◽  
T Burnouf ◽  
C V Prowse
Keyword(s):  
High Purity ◽  
Time Course ◽  
Prothrombin Complex Concentrate ◽  
Degradation Products ◽  
Canine Model ◽  
Factor Ix ◽  
In Vitro Tests ◽  
Albumin Infusion

SummaryA non-stasis canine model of thrombogenicity has been used to evaluate batches of high purity factor IX concentrates from 4 manufacturers and a conventional prothrombin complex concentrate (PCC). Platelets, activated partial thromboplastin time (APTT), fibrinogen, fibrin(ogen) degradation products and fibrinopeptide A (FPA) were monitored before and after infusion of concentrate. Changes in FPA were found to be the most sensitive and reproducible indicator of thrombogenicity after infusion of batches of the PCC at doses of between 60 and 180 IU/kg, with a dose related delayed increase in FPA occurring. Total FPA generated after 100-120 IU/kg of 3 batches of PCC over the 3 h time course was 9-12 times that generated after albumin infusion. In contrast the amounts of FPA generated after 200 IU/kg of the 4 high purity factor IX products were in all cases similar to albumin infusion. It was noted that some batches of high purity concentrates had short NAPTTs indicating that current in vitro tests for potential thrombogenicity may be misleading in predicting the effects of these concentrates in vivo.

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