Electrogenic, K + -dependent chloride transport in locust hindgut

Potassium chloride is the major salt recycled in most insect excretory systems. Ion and water reabsorption occur in the rectum by active transport of Cl- and largely passive movement of K+. Both these processes are stimulated several fold by a neuropeptide hormone acting via cyclic AMP (cAMP). This Cl- transport process was investigated by using intracellular ion-sensitive microelectrodes, radiotracer flux measurements, voltage clamping, ion substitutions and inhibitors. The mucosal entry step for Cl- is energy-requiring and highly selective, and is stimulated directly by cAMP and luminal K +. Under some experimental conditions, measured electrochemical potentials for cations across the mucosal membrane are too small to drive Cl- entry by NaCl or KC1 cotransport mechanisms; moreover, net 36C1- flux is independent of the apical Na+ potential. Similarly no evidence for a HCO 3 -Cl- exchange was obtained. We conclude that Cl- transport in locust gut is different from mechanisms currently proposed for vertebrate tissues.

1981 ◽  
Vol 90 (1) ◽  
pp. 123-142
Author(s):  
M. M. P. RAMOS ◽  
J. C. ELLORY

1. The tissue was found to have a serosa negative potential, and short-circuit currents equivalent to the net Cl transport. 2. A significant part of the Cl uptake was Na dependent and a similar fraction of the Na uptake was Cl dependent. 3. Short-circuit current and uptake of both ions were inhibited by loop diuretics and analogues. 4. I80 and P.D. were abolished by ouabain. 5. The observations are consistent with the idea of a coupled NaCl entry into the cell, using the energy inherent in the Na gradient; Na being pumped out of the cells by the Na pump and followed electrically by Cl−. Net chloride transport and the serosa negative potential would be a consequence of the permselective properties of the junctions allowing Na but not Cl to recycle back to the mucosal solution.


1983 ◽  
Vol 244 (2) ◽  
pp. R131-R142 ◽  
Author(s):  
J. Hanrahan ◽  
J. E. Phillips

The rectum is the main reabsorptive site in the excretory system of locusts. The primary urine entering this organ from the Malpighian tubules is rich in K+ (140 mM) and Cl- (90 mM), and most of this fluid is normally reabsorbed. Fluid and active Cl- reabsorption in the rectum are regulated by neuropeptide hormones from the corpus cardiacum. We have studied the mechanism of KCl reabsorption using voltage clamp, tracers, double-barreled ion-sensitive microelectrodes, and ion substitutions. Locust Cl- absorption differs from vertebrate systems in that it is not dependent on Na+ or HCO-3/CO2, and it is insensitive to normal inhibitors of Cl- transport. Entry of Cl- into rectal cells is active, electrogenic, and stimulated by luminal K+. This cation substantially increases the electrochemical gradient across the apical membrane against which Cl- is pumped; therefore K+ does not act solely and indirectly by electrical coupling. Kinetic studies also suggest that K+ activates the Cl- pump. Consequently at least two levels of control are exerted during cAMP stimulation; K+ permeability of the epithelium and the transepithelial potential generated by active Cl- transport both increase. The enhanced net K+ absorption from the lumen side after stimulation is largely passive, being electrically coupled to Cl- transport. However, this general increase in KCl absorption is "fine tuned" by K+ itself, through its direct effect on the Cl- pump.


1976 ◽  
Vol 3 (4) ◽  
pp. 443
Author(s):  
W.J Lucas ◽  
F.A Smith

Parallel studies on the influence of irradiance on net H+ efflux and *36Cl- uptake were conducted on C. corallina. Following the dark-to-light transition, a lag period of 8-15 min was observed before net H+ efflux activity could be discerned experimentally. Decreasing the irradiance did not significantly lengthen this lag period. Studies on *36Cl- uptake revealed that a lag period of 40-60 min was required before the Cl- transport system attained maximum activity, governed by the prevailing experimental conditions. These results are discussed in relation to the Cl- transport hypotheses proposed by Spear et al. and by Smith. It would seem that the hypothesis of Spear et al., in its present form, is invalid for Chara corallina. The results were inconclusive in terms of support (or otherwise) for a Cl-/OH- antiporter. However, the Cl-/OH- antiporter hypothesis originally proposed by Smith will require significant modification, especially in terms of the energy supply.


1982 ◽  
Vol 243 (2) ◽  
pp. F167-F172 ◽  
Author(s):  
M. Imai ◽  
E. Kusano

Arginine vasopressin (AVP) has been shown to stimulate active Cl transport across the medullary thick ascending limb of Henle's loop (MAL) in association with an increase in adenylate cyclase activity. To determine whether the failure to demonstrate active Cl transport across the thin ascending limb of Henle's loop (TAL) in previous in vitro perfusion studies was due to the absence of AVP in the preparation, we examined the effect of AVP on adenylate cyclase activity and Cl transport in the hamsters TAL. AVP (1 mU/ml) increased adenylate cyclase activity in the hamster TAL (20.7 +/- 5.2 control vs. 46.2 +/- 10.1 fmol . mm-1 . 30 min-1, n = 6, P less than 0.05) but not in the descending limb (27.8 +/- 7.0 control vs. 20.4 +/- 2.7, n = 4, P less than 0.05). When both MAL and TAL were perfused, a lumen-positive transepithelial voltage (Vt) was observed. The Vt was increased by adding 1 or 10 mU/ml AVP to the bath. When only the TAL was perfused, the Vt was not different from zero. Similar results were obtained in mouse renal tubules. In other experiments, AVP did not affect the diffusion potential generated when a transepithelial NaCl gradient was present. AVP or dibutyryl cAMP caused little or no change in efflux of radioactive chloride across the hamster TAL. These findings suggest that electrogenic chloride transport is not demonstrable in the TAL even in the presence of AVP. The physiologic role of AVP-sensitive adenylate cyclase in the TAL remains to be established.


1988 ◽  
Vol 8 (1) ◽  
pp. 27-33 ◽  
Author(s):  
M. A. McPherson ◽  
D. K. Shori ◽  
R. L. Dormer

A biochemical link is proposed between recent observations on defective regulation of Cl− transport in CF respiratory epithelial cells and studies showing altered biological activity of calmodulin in exocrine glands from CF patients. A consensus is emerging that defective β-adrenergic secretory responsiveness in CF cells is caused by a defect in a regulator protein at a site distal to cyclic AMP formation. Our results indicate that this protein might be a specific calmodulin acceptor protein which modifies the activity of calmodulin in epithelial cells. Alteration in Ca2+/calmodulin dependent regulation of Cl− transport and protein secretion could explain (i) alterations in Ca2+ homeostasis seen in CF, (ii) defective β-adrenergic responses of CF cells, and (iii) the observed inability of cyclic AMP (acting via its specific protein kinase, A-kinase) to open apical membrane Cl− channels in CF epithelial cells. Most of the physiological abnormalities of CF including elevated sweat electrolytes and hyperviscous mucus can be explained on this basis.


2002 ◽  
Vol 205 (11) ◽  
pp. 1645-1655 ◽  
Author(s):  
Juan P. Ianowski ◽  
Robert J. Christensen ◽  
Michael J. O'Donnell

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.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Iga Hołyńska-Iwan ◽  
Karolina Szewczyk-Golec

Abstract The measurement of electric potential and resistance reflect the transport of sodium and chloride ions which take place in keratinocytes and is associated with skin response to stimuli arising from external and internal environment. The aim of the study was to assess changes in electrical resistance and the transport of chloride and sodium ions, under iso-osmotic conditions and following the use of inhibitors affecting these ions’ transport, namely amiloride (A) and bumetanide (B). The experiment was performed on 104 fragments of rabbit skin, divided into three groups: control (n = 35), A—inhibited sodium transport (n = 33) and B—inhibited chloride transport (n = 36). Measurement of electrical resistance (R) and electrical potential (PD) confirmed tissue viability during the experiment, no statistically significant differences in relation to control conditions were noted. The minimal and maximal PD measured during stimulation confirmed the repeatability of the recorded reactions to the mechanical and mechanical–chemical stimulus for all examined groups. Measurement of PD during stimulation showed differences in the transport of sodium and chloride ions in each of the analyzed groups relative to the control. The statistical analysis of the PD measured in stationary conditions and during mechanical and/or mechanical–chemical stimulation proved that changes in sodium and chloride ion transport constitute the physiological response of keratinocytes to changes in environmental conditions for all applied experimental conditions. Assessment of transdermal ion transport changes may be a useful tool for assessing the skin condition with tendency to pain hyperactivity and hypersensitivity to xenobiotics.


1978 ◽  
Vol 72 (2) ◽  
pp. 203-218 ◽  
Author(s):  
T E Ryan ◽  
C E Barr ◽  
J P Zorn

Transmembrane movements of K+ and Cl- were studied under a variety of experimental conditions. Potassium was found to carry more than 50% of an externally applied inward positive current. The increase in K+ influx was much greater than that predicted by the purely passive model. The increase in Cl- efflux accounted for less than 10% of the applied current, in agreement with the value predicted for passive movement. 2,4-Dinitrophenol (DNP) caused an 80% reduction in K+ transference and a corresponding increase in the measured electrical resistance of the membrane. DNP also reduced the isotopically measured resting K+ influx and caused a substantial increase in both Cl- influx and efflux. Lowering of the pH from 5.7 to 4.7 also reduced the net K+ influx but without drastically altering the membrane resistance. It appears the major portion of an externally applied current does not travel through passive channels, but rather is shunted through a different membrane component. In conjunction with evidence previously establishing the H+ pump as the primary ion pump in Nitella, the data presented here are consistent with a K+/H+ exchange mechanism which can account for the observed net K+ accumulation and maintenance of the membrane potential above the electrochemical equilibrium potential of the major ions. This mechanism appears to be a likely candidate for the current shunt.


1976 ◽  
Vol 68 (6) ◽  
pp. 583-599 ◽  
Author(s):  
P C Brazy ◽  
R B Gunn

The chloride self-exchange flux across the human red cell membrane is rapidly and reversibly inhibited by 10(-4) M furosemide, a potent chloruretic agent. Furosemide reduces the chloride flux at all chloride concentrations and increases the cellular chloride concentration at which the flux is half-maximum. Kinetic analysis of the flux measurements made at several furosemide and chloride concentrations yields a pattern of mixed inhibition with a dissociation constant for the inhibitor-transport mechanism complex of 5 X 10(-5) M. From this pattern of inhibition and other observations, including that the percent inhibition is independent of pH (range 5.6-8.9), we conclude that the anionic form of furosemide interacts primarily with the chloride transport mechanism at a site separate from both the transport site and the halide-reactive modifier site.


1995 ◽  
Vol 198 (10) ◽  
pp. 2237-2244 ◽  
Author(s):  
F Jensen ◽  
J Brahm

The continuous flow tube method was used to investigate the kinetics of chloride transport, and its potential oxygenation-dependency, in red blood cells (RBCs) from four teleost fish species and man. A significant interspecific variation in Cl- transport kinetics was found. At 15 °C, the rate constant k for unidirectional 36Cl- efflux was significantly lower in RBCs from eel and carp than in RBCs from rainbow trout and Atlantic cod. The values of k of cod RBCs at 15 °C and of human RBCs at 37 °C were not significantly different. The volume and surface area of the RBCs were evaluated and used to calculate the apparent membrane permeability to Cl- (PCl). The magnitude of PCl followed the sequence: eel<carp<trout¾cod. PCl values in trout and cod at 15 °C were similar to human values at 37 °C. An extrapolation of human values to 15 °C revealed that the Cl- shift at this temperature was considerable faster in all four teleosts than in man. This illustrates appropriate adaption of band-3-mediated anion transport to the different temperature regimes encountered by fish and mammals. The Cl- transport kinetics did not differ significantly between oxygenated and deoxygenated RBCs in any of the species examined. The apparent absence of any effect of a change in haemoglobin oxygen-saturation may be related to the presence of a flexible link which results in minimal interaction between the membrane domain (mediating Cl- transport) and the cytoplasmic domain (to which oxygenation-dependent haemoglobin binding occurs) of band 3. In carp, Cl- transport kinetics were not influenced by pH over the extracellular pH (pHe) range 7.6­8.36, which spans the in vivo pHe range. The data are discussed in relation to the rate-limiting role of red blood cell HCO3-/Cl- exchange for CO2 excretion.


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