Potassium channels in the basolateral membrane of the rectal gland ofSqualus acanthias

1987 ◽  
Vol 409 (1-2) ◽  
pp. 107-113 ◽  
Author(s):  
Heinz Gögelein ◽  
Rainer Greger ◽  
Eberhard Schlatter
Keyword(s):  
Potassium Channels ◽  
Basolateral Membrane ◽  
Rectal Gland

Immunocytochemical studies of the Na-K-Cl cotransporter of shark kidney

1996 ◽  
Vol 270 (6) ◽  
pp. F927-F936 ◽  
Author(s):  
D. Biemesderfer ◽  
J. A. Payne ◽  
C. Y. Lytle ◽  
B. Forbush
Keyword(s):  
Plasma Membrane ◽  
Immunoelectron Microscopy ◽  
Basolateral Membrane ◽  
Distal Tubule ◽  
Rectal Gland ◽  
Apical Plasma Membrane ◽  
Secretory Cells ◽  
Weak Staining ◽  
Basolateral Plasma Membrane ◽  
Kidney Functions

The Na-K-Cl cotransporter (NKCC or BSC) has been described in numerous secretory and reabsorptive epithelia and is an important part of the mechanism of NaCl reabsorption in both the mammalian and elasmobranch kidneys. We have recently developed a panel of four monoclonal antibodies (MAbs) raised to the 195-kDa Na-K-Cl cotransport protein of the shark rectal gland (sNKCC1), which is expressed along the basolateral plasma membrane of secretory cells in this tissue (29). Here, we report immunologic studies of the Na-K-Cl cotransporter in the kidney of the dogfish shark Squalus acanthias. Western blot analysis of shark renal microsomes using MAbs J3, J7, and J25 identified proteins of approximately 195 and 150 kDa, whereas MAb J4 was not reactive. To define the cellular and subcellular distribution of the cotransport protein, immunofluorescence and immunoelectron microscopy studies were performed on fixed kidneys. Immunofluorescence microscopy on semithin (0.5-micron) cryosections demonstrated that MAbs J3, J7, and J25 intensely stained the apical plasma membrane of all distal tubule segments. Weak staining was also seen along the basolateral membrane of most distal nephrons. Immunoelectron microscopy confirmed this observation and showed that some of these segments were morphologically similar to diluting segments from other species. MAbs also reacted with the brush border and, to a lesser extent, the basolateral membrane of proximal tubules. This study supports the hypothesis that the lateral bundle zone of the elasmobranch kidney functions as a countercurrent exchanger and is consistent with the presence of multiple isoforms of the Na-K-Cl cotransporter in the shark kidney.

Selective permeability barrier to urea in shark rectal gland

2005 ◽  
Vol 289 (1) ◽  
pp. F83-F89 ◽  
Author(s):  
Joshua D. Zeidel ◽  
John C. Mathai ◽  
John D. Campbell ◽  
Wily G. Ruiz ◽  
Gerard L. Apodaca ◽  
...  
Keyword(s):  
Activation Energy ◽  
Epithelial Cells ◽  
Water Flow ◽  
Water Permeability ◽  
Apical Membrane ◽  
Basolateral Membrane ◽  
Water Flux ◽  
Rectal Gland ◽  
Basolateral Membranes ◽  
Urea Permeability

Elasmobranchs such as the dogfish shark Squalus acanthius achieve osmotic homeostasis by maintaining urea concentrations in the 300- to 400-mM range, thus offsetting to some degree ambient marine osmolalities of 900–1,000 mosmol/kgH2O. These creatures also maintain salt balance without losing urea by secreting a NaCl-rich (500 mM) and urea-poor (18 mM) fluid from the rectal gland that is isotonic with the plasma. The composition of the rectal gland fluid suggests that its epithelial cells are permeable to water and not to urea. Because previous work showed that lipid bilayers that permit water flux do not block flux of urea, we reasoned that the plasma membranes of rectal gland epithelial cells must either have aquaporin water channels or must have some selective barrier to urea flux. We therefore isolated apical and basolateral membranes from shark rectal glands and determined their permeabilities to water and urea. Apical membrane fractions were markedly enriched for Na-K-2Cl cotransporter, whereas basolateral membrane fractions were enriched for Na-K-ATPase. Basolateral membrane osmotic water permeability (Pf) averaged 4.3 ± 1.3 × 10−3 cm/s, whereas urea permeability averaged 4.2 ± 0.8 × 10−7 cm/s. The activation energy for water flow averaged 16.4 kcal/mol. Apical membrane Pf averaged 7.5 ± 1.6 × 10−4 cm/s, and urea permeability averaged 2.2 ± 0.4 × 10−7 cm/s, with an average activation energy for water flow of 18.6 kcal/mol. The relatively low water permeabilities and high activation energies argue strongly against water flux via aquaporins. Comparison of membrane water and urea permeabilities with those of artificial liposomes and other isolated biological membranes indicates that the basolateral membrane urea permeability is fivefold lower than would be anticipated for its water permeability. These results indicate that the rectal gland maintains a selective barrier to urea in its basolateral membranes.

Cl- secretion by cultured shark rectal gland cells. II. Effects of forskolin on cellular electrophysiology

1991 ◽  
Vol 260 (4) ◽  
pp. C824-C831 ◽  
Author(s):  
W. M. Moran ◽  
J. D. Valentich
Keyword(s):  
Apical Membrane ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Ringer Solution ◽  
Rectal Gland ◽  
Electrical Potential Difference ◽  
Electrophysiological Properties ◽  
Shark Rectal Gland ◽  
Membrane Electrical Potential ◽  
Cl Conductance

Employing microelectrode techniques we have assessed the cellular electrophysiological properties of shark rectal gland (SRG) cells in primary culture. In the absence of secretagogues a 10-fold reduction in the Cl- concentration of the apical superfusate shark Ringer solution had little effect on either apical membrane electrical potential difference (Va) or fractional resistance (fRa), indicating little, if any, apical membrane Cl- conductance. Superfusing the basolateral surface with high-K+ shark Ringer solution (K+ increased 10-fold) depolarized the basolateral membrane electrical potential difference (Vb) by 43 mV, indicating that this barrier is largely K+ conductive. In addition, basolateral Ba2+ (5 mM) depolarized Vb by 12 mV and reduced fRa from 0.92 to 0.58, results consistent with a K(+)-conductive basolateral membrane in unstimulated SRG cells. Basolateral forskolin (10(-6) M) depolarized Va by 25 mV and caused a dramatic reduction in fRa from 0.97 to approximately 0.10. Under these conditions, a 10-fold decrease in apical superfusate Cl- concentration depolarized Va by 37 mV, revealing an adenosine 3',5'-cyclic monophosphate-induced apical membrane Cl- conductance. The time course of the forskolin-induced changes in Va and Vb suggests that the basolateral membrane K+ conductance increased and maintained the driving force for apical Cl- exit, as in other Cl(-)-secreting epithelia. These electrophysiological properties compare favorably with those of the perfused SRG tubule and indicate that SRG primary cultures are a suitable model for Cl(-)-secreting epithelia.

Na-K-Cl cotransport in the shark rectal gland. I. Regulation in the intact perfused gland

1992 ◽  
Vol 262 (4) ◽  
pp. C1000-C1008 ◽  
Author(s):  
B. Forbush ◽  
M. Haas ◽  
C. Lytle
Keyword(s):  
Time Course ◽  
Loop Diuretic ◽  
Basolateral Membrane ◽  
Ringer Solution ◽  
K Channel ◽  
Rectal Gland ◽  
Cotransport System ◽  
Nacl Secretion ◽  
Dogfish Shark ◽  
Perfusion Period

To investigate regulation of the Na-K-Cl cotransport system in the rectal gland of the dogfish shark Squalus acanthias, we examined binding of the loop diuretic [3H]benzmetanide to the intact gland. Glands were perfused with a shark Ringer solution, either in a basal state or stimulated with vasoactive intestinal peptide (VIP). [3H]benzmetanide was added to the perfusion solution for the last 25 min of perfusion, after which the gland was homogenized and the amount of bound [3H]benzmetanide was determined in the membrane fraction. Most of the membrane-associated [3H]-benzmetanide appeared to be associated with the Na-K-Cl cotransporter as judged by the dissociation rates at 0 degree C and 20 degrees C, by labeling with a photosensitive analogue, and by continued association of [3H]benzmetanide with membrane protein on solubilization. With the use of [3H]4-benzoyl-5-sulfamoyl-3-(3- thenyloxy)benzoic acid, a photosensitive analogue of benzmetanide, a 200-kDa protein was selectively labeled on exposure to ultraviolet light. It was also possible to detect [3H]-benzmetanide binding during the perfusion period as an arterial-venous difference, thereby providing a time course of the binding process. In comparing two groups of five glands each, VIP stimulated NaCl secretion 20-fold and [3H]benzmetanide binding 16-fold, providing strong evidence that the Na-K-Cl cotransport system is activated as part of the process of stimulation of secretion. The VIP-stimulated increase in [3H]benzmetanide binding was completely inhibited when Ba was added to the perfusate to block K channel-mediated K exit across the basolateral membrane.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between sodium transport and intracellular ATP in isolated perfused rabbit proximal convoluted tubule

1991 ◽  
Vol 261 (4) ◽  
pp. F634-F639 ◽  
Author(s):  
J. S. Beck ◽  
S. Breton ◽  
H. Mairbaurl ◽  
R. Laprade ◽  
G. Giebisch
Keyword(s):  
Potassium Channels ◽  
Cell Volume ◽  
Intracellular Ph ◽  
Sodium Transport ◽  
Sodium Pump ◽  
Basolateral Membrane ◽  
Potassium Conductance ◽  
Transference Number ◽  
Proximal Convoluted Tubule ◽  
A Cell

The effect of alterations in sodium transport on cell ATP content and pH in the isolated perfused proximal convoluted tubule (PCT) of the rabbit was examined. Stimulating sodium transport by the addition of luminal glucose and alanine decreased cell ATP from 4.44 +/- 0.93 to 2.69 +/- 0.62 mM (n = 4), increased intracellular pH by 0.13 +/- 0.02 (n = 7), and increased cell volume by 0.10 +/- 0.02 nl/mm (n = 4). Blocking the sodium pump with 10(-4) M strophanthidin in tubules in which sodium transport had been stimulated increased cell ATP from 2.04 +/- 0.24 to 2.42 +/- 0.32 mM (n = 6). In parallel experiments the same dose of strophanthidin depolarized the basolateral membrane from -52.6 +/- 1.9 to -6.4 +/- 1.6 mV, depolarized the transepithelial potential from -3.2 +/- 0.3 to -0.1 +/- 0.1 mV, and reduced the basolateral membrane potassium transference number from 0.47 to 0.26 indicating a reduction in basolateral potassium conductance. Since strophanthidin caused a cell alkalinization of 0.15 +/- 0.03, this latter effect cannot be due to changes of intracellular pH. Strophanthidin caused no change in cell volume over the period studied, suggesting that stretch-activated potassium channels are not involved either. Instead, potassium conductance inhibition may be the result of the closure of ATP-sensitive potassium channels. These same channels might thus be partly responsible for the increase in potassium conductance commonly observed during stimulation of sodium transport.

Intracellular chloride activities in the isolated perfused shark rectal gland

1983 ◽  
Vol 245 (5) ◽  
pp. F640-F644
Author(s):  
M. J. Welsh ◽  
P. L. Smith ◽  
R. A. Frizzell
Keyword(s):  
Basolateral Membrane ◽  
Apical Cell ◽  
Electrical Potential ◽  
Electrochemical Potential ◽  
Potential Difference ◽  
Rectal Gland ◽  
Electrical Potential Difference ◽  
Apical Cell Membrane ◽  
Shark Rectal Gland ◽  
Electrochemical Equilibrium

The isolated, perfused shark rectal gland secretes Cl when stimulated with adenosine 3',5'-cyclic monophosphate (cAMP). To investigate the mechanism of secretion, we used Cl-selective and conventional (KCl-filled) microelectrodes to measure the intracellular Cl activity (aClc). Under nonsecreting conditions, the electrical potential difference across the basolateral membrane (psi b) was -78 m V and aClc was 57 mM, a value seven times greater than predicted for electrochemical equilibrium across the basolateral membrane. When theophylline and 8-bromo-cAMP were added to the perfusate, the transglandular electrical potential difference doubled and the rate of fluid secretion increased 20-fold; however, neither psi b nor aClc changed. During both nonsecreting and secreting conditions the intracellular accumulation of Cl results in an electrochemical potential difference favoring Cl exit across the apical cell membrane. The constancy of aClc despite the variation in secretion rate suggests that stimulation is associated with an equivalent enhancement of net Cl movement across both the apical and basolateral membranes. When stimulated glands were perfused with Na-free (choline) Ringer, secretion was abolished and aClc fell toward the value predicted for electrochemical equilibrium. These findings suggest that the "uphill" step in Cl secretion lies at the basolateral membrane, where cellular Cl accumulation probably involves secondary active transport; i.e., Cl entry is driven by an inwardly directed electrochemical potential difference for Na.

Morphology of the rectal gland of the spiny dogfish (Squalus acanthias) shark in response to feeding

2009 ◽  
Vol 87 (5) ◽  
pp. 440-452 ◽  
Author(s):  
Victoria Matey ◽  
Chris M. Wood ◽  
W. Wesley Dowd ◽  
Dietmar Kültz ◽  
Patrick J. Walsh
Keyword(s):  
Early Stage ◽  
Basolateral Membrane ◽  
Small Diameter ◽  
Squalus Acanthias ◽  
Rectal Gland ◽  
Secretory Cells ◽  
Spiny Dogfish ◽  
Apical Surface ◽  
Stratified Epithelium ◽  
Outer Capsule

The morphology of the rectal gland was examined in spiny dogfish ( Squalus acanthias L, 1758) sharks fasted (1 week) or 6 and 20 h postfeeding. The morphology of the fasted gland showed a pattern reflecting a dormant physiology, with thick gland capsule, thick stratified epithelium, and secretory parenchyma with tubules of small diameter and lumen. The secretory cells of the tubular epithelium were enlarged and irregularly shaped with abnormally condensed or highly vacuolized cytoplasm containing numerous lysosomes. Early-stage apoptotic cells were not uncommon. Secretory cells showed signs of low activity, e.g., mitochondria with weakly stained matrix and small cristae, poorly branched infoldings of basolateral membranes, and microvesicle-free subapical cytoplasm. All characteristics examined changed significantly upon feeding, consistent with increased salt and fluid secretion: the outer capsule muscle layer and the stratified epithelium decreased in diameter; the tubules enlarged; the secretory cells showed extensive development of the basolateral membrane, more mitochondria, and abundant apical microvesicles. Secretory cell apical surface was increased. The minor differences between morphology in 6 and 20 h postfeeding indicated that changes took place rapidly and were complete by 6 h. Our results are discussed in the context of prior studies of metabolism, proteomics, and cellular pathways of gland activation.

Regulation of MRP2-mediated transport in shark rectal salt gland tubules

2002 ◽  
Vol 282 (3) ◽  
pp. R774-R781 ◽  
Author(s):  
David S. Miller ◽  
Rosalinde Masereeuw ◽  
Karl J. Karnaky
Keyword(s):  
Protein Kinase ◽  
Receptor Antagonist ◽  
Efflux Pump ◽  
Basolateral Membrane ◽  
Salt Gland ◽  
Selective Inhibitor ◽  
Rectal Gland ◽  
Fluorescent Substrate ◽  
Shark Rectal Gland ◽  
Sulforhodamine 101

We examined endothelin-1 (ET-1) regulation of the xenobiotic efflux pump, multidrug resistance-associated protein isoform 2 (MRP2), in intact dogfish shark rectal salt gland tubules using a fluorescent substrate sulforhodamine 101 and confocal microscopy. Subnanomolar to nanomolar concentrations of ET-1 rapidly reduced the cell-to-lumen transport of sulforhodamine 101. These effects were prevented by an ETBreceptor antagonist but not by an ETA receptor antagonist. Immunostaining with an antibody to mammalian ETB receptors showed specific localization to the basolateral membrane of the shark rectal gland epithelial cells. ET-1 effects on transport were blocked by a protein kinase C (PKC)-selective inhibitor, implicating PKC in ET-1 signaling. A protein kinase A (PKA)-selective inhibitor had no effect. Forskolin reduced luminal accumulation of sulforhodamine 101, but inhibition of PKA did not block the forskolin effect. Consistent with this observation, a cAMP analog that does not activate PKA reduced luminal accumulation of sulforhodamine 101. These results indicate that shark rectal gland transport on MRP2 is regulated by ET acting through an ETB receptor and PKC. In addition, cAMP affects transporter function through a PKA-independent mechanism, possibly by competition for transport.

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