Diluting segment in kidney of dogfish shark. II. Electrophysiology of apical membranes and cellular resistances.

Diluting segments from the bundle zone of the dogfish shark kidney were perfused in vitro and the electrophysiological characteristics of this segment investigated using conventional microelectrodes and cable analysis. In 21 tubules perfused with symmetrical Ringer solutions the average transepithelial voltage (Vte), transepithelial conductance (Gte), and equivalent short circuit current (Isc) were 8.7 +/- 0.

Diluting segment in kidney of dogfish shark. I. Localization and characterization of chloride absorption.

Single tubules, dissected from the peritubular sheath of the dorsal bundle zone of kidney of the dogfish shark, Squalus acanthias, were perfused in vitro at 17-18 degrees C. This segment is the largest of the five in the peritubular sheath and had average inner and outer diameters of 46.9 +/- 1.

Rapid hypertonic cell volume regulation in the perfused inner medullary collecting duct.

Differential interference contrast microscopic images were used to assess the cell volume regulatory increase (VRI) response of rat IMCD segments isolated from the mid-inner medullary region of pathogen-free Sprague-Dawley rats and perfused in vitro at 37 degrees C. In the absence of ADH. IMCD cells behaved in an osmometric fashion over the range of extracellular osmolalities 290 to 386 mOsm/kg H2O and had an osmotic space equal to 54.

Sodium-coupled ion cotransport and the volume regulatory increase response.

In conclusion, maintenance of volume homeostasis is a fundamental requirement of all cells. For many cell types, this process requires expression of ion cotransport mechanisms as well as accumulation of osmotically-active organic compounds. Recent observations have indicated that the cellular mechanisms responsible for modulating hypertonic volume regulation are complex and appear to involve hormonal, biochemical and physico-chemical stimuli.

Lymphocyte subset analysis to predict progression to AIDS in a cohort of homosexual men in San Francisco.

A group of 10 leukocyte and lymphocyte subsets were measured by simultaneous dual immunofluorescence and flow cytometry in a group of homosexual men from the San Francisco General cohort. Absolute numbers and percentages of lymphocytes were determined in 30 individuals who progressed to AIDS and 29 who did not over a 44-month period at annual intervals. At entry into the study, all subjects were asymptomatic, HIV seropositive, and had multiple changes in lymphocyte subsets compared to HIV-negative controls.

Cell membranes impermeable to NH3.

Classically, there is a direct correlation between the lipophilic nature of a molecule and its rate of permeation across a biological membrane, so cell membranes should be more permeable to small, neutral molecules than they are to charged molecular species of similar size. Consequently, the distribution of NH+4 in biological systems is generally believed to be due to the rapid diffusion and equilibration of lipophilic NH3 across cell membranes and the accumulation of NH+4 to be governed by pH differences between compartments. Here we report that renal tubule cells from the medullary thick ascending limb of Henle have an apical membrane which is not only virtually impermeable to NH3, but is also highly permeable to NH+4.

Organic osmolytes in inner medulla of Brattleboro rat: effects of ADH and dehydration.

Inner medullary methylamine [glycerophosphorylcholine (GPC) and glycine betaine (betaine)] and polyol [sorbitol and myo-inositol (inositol)] osmolytes were measured in water-restricted and antidiuretic hormone (ADH)-infused Brattleboro (DI) rats. Compared with DI rats allowed water ad libitum, rats dehydrated for 3 days had higher urinary osmolality (Uosmol) (812 vs. 239 mosmol/kgH2O) and plasma osmolality (Posmol) (333 vs.

Renal inner medullary choline dehydrogenase activity: characterization and modulation.

Betaine belongs to the trimethylamine class of osmolytes (osmotically active substances believed to play an important role in cell volume homeostasis) and has recently been identified in the inner medulla of the mammalian kidney. Trimethylamines accumulate in the renal inner medulla during hypertonic stress, and betaine content in the inner medulla has been shown recently to increase during hypernatremia, yet the mechanisms governing the modulation of trimethylamine content and, in particular, of betaine content are not well understood. In this study, we demonstrate the presence of choline dehydrogenase activity in the renal inner medullas of three separate rat strains.

Hypotonic cell volume regulation in mouse medullary thick ascending limb: effects of ADH.

Differential interference contrast microscopy was used in combination with standard electrophysiological techniques in the in vitro perfused mouse medullary thick ascending limb of Henle's loop (MAL) to evaluate the cell volume responses of this nephron segment during and following exposure to hypotonic media and to assess the role of antidiuretic hormone (ADH) and net salt absorption on the associated volume regulatory processes. Reductions in extracellular osmolality by 50 mosmol resulted in rapid increases in cell volume of approximately 20% with or without exposure to ADH. Cell volume recovery (volume-regulatory decrease, VRD) was much slower in the presence, than in the absence, of ADH.

Accumulation of major organic osmolytes in rat renal inner medulla in dehydration.

Osmotically active organic solutes, osmolytes, exist at high concentrations in the renal inner medulla; however, their modulation during antidiuresis remains largely undefined. Renal osmolyte levels were measured by nuclear magnetic resonance spectroscopy and biochemical assays in perchloric acid extracts from normal and dehydrated (3 days) rats. Dehydration increased urine osmolality from 1,503 to 3,748 mosmol/kg and inner medullary urea content from 2,036 to 4,405 nmol/mg protein.

Modulation of Na-K-ATPase activity in the mouse medullary thick ascending limb of Henle. Effects of mineralocorticoids and sodium.

This study investigates the effect of variations in mineralocorticoid as well as cell sodium delivery and uptake on Na-K-ATPase activity in the mouse medullary thick ascending limb of Henle (mTALH). Pharmacologic doses of the mineralocorticoid deoxycorticosterone acetate (DOCA) resulted in a 28% increase of Na-K-ATPase activity. Furosemide-induced inhibition of sodium uptake by the mTALH cell also resulted in Na-K-ATPase activity reduction (45%).

A simple method for continuous measurement of the volume of cells in suspension.

A simple electrical method for continuous monitoring of the volume of cells in suspension is described. The procedure consists of placing cells in a medium containing an impermeant quaternary ammonium ion, such as tetramethylammonium (TMA), which behaves as an extracellular volume marker, and monitoring the activity of this ion with a quaternary ammonium ion-specific minielectrode. Changes in the activity of the cell impermeant TMA ion in the extracellular medium quantitatively reflect movements of water into, or out of, cells and, therefore, provide a direct measure of alterations in cell volume.

Uninephrectomy and dietary protein affect fluid absorption in rabbit proximal straight tubules.

To further delineate the mechanisms involved in the renal compensatory response to renal mass ablation, we evaluated the interactions of protein intake and uninephrectomy on total, active, and passive fluid absorption (Jv) in the in vitro perfused superficial proximal straight tubule (SFPST) isolated from sham-operated or uninephrectomized rabbits pair fed 7, 14, or 45% protein diets. In all protein groups, we observed significant increases in Jv and in cell height from SFPST segments obtained from remnant renal tissue harvested 3 wk post-uninephrectomy when compared with sham-operated controls. Uninephrectomy also led to compensatory hypertrophy of whole-kidney mass and to elevations in inulin clearances.

ADH increases apical Na+, K+, 2Cl- entry in mouse medullary thick ascending limbs of Henle.

These studies were designed to evaluate the mechanism for the ADH-dependent increase in transcellular conductance (Gc, mS X cm-2), which accompanies hormone-dependent increases in the spontaneous transepithelial voltage (Ve, mV) and in the net rate of Cl- absorption in single medullary thick ascending limbs of Henle (mTALH) isolated from mouse kidney. The total transepithelial conductance (Ge, mS X cm-2) was measured with perfusing solutions containing 5 mM K+, zero Ba2+; Gc was that component of Ge blocked by luminal 20 mM Ba2+, zero K+. In paired experiments, antidiuretic hormone (ADH) increased Gc from 44.

Hypertonic cell volume regulation in mouse thick limbs. II. Na+-H+ and Cl(-)-HCO3- exchange in basolateral membranes.

Differential interference contrast microscopy and standard electrophysiological techniques were used to evaluate the transport processes involved in antidiuretic hormone (ADH)-dependent hypertonic cell volume regulation in the in vitro perfused mouse medullary thick ascending limb of Henle. Hypertonic cell volume regulation appeared to involve NaCl uptake into cells, since the cell volume increase after osmotic shrinkage in hypertonic media could be abolished either by symmetrical removal of Na+ from external solutions or by bath Cl- omission. The volume-regulatory process also required CO2/HCO3- in external media and could be abolished by the lipophilic carbonic anhydrase inhibitor, ethoxzolamide, in the presence of CO2/HCO3-.

Hypertonic cell volume regulation in mouse thick limbs. I. ADH dependency and nephron heterogeneity.

Differential interference contrast microscopy was used in combination with standard electrophysiological techniques in the in vitro perfused mouse medullary (mTALH) and cortical (cTALH) thick ascending limbs of Henle to evaluate the cell volume responses of these nephron segments to sudden increases in peritubular osmolality and to assess the role of antidiuretic hormone (ADH) and net NaCl absorption on hypertonic volume regulation. In the absence of CO2/HCO3- in external media, the cells of the mTALH behaved in a simple osmometric fashion, with an osmotic space equivalent to 70-80% of the total cell volume. However, in CO2/HCO3- -containing media, the cells of the mTALH, but not the cTALH, were able to increase their cell volume to the original volume after shrinkage in peritubular media made hypertonic with either NaCl or mannitol.

Ionic conductance pathways in the mouse medullary thick ascending limb of Henle. The paracellular pathway and electrogenic Cl- absorption.

Net Cl- absorption in the mouse medullary thick ascending limb of Henle (mTALH) involves a furosemide-sensitive Na+:K+:2 Cl- apical membrane symport mechanism for salt entry into cells, which occurs in parallel with a Ba++-sensitive apical K+ conductance. The present studies, using the in vitro microperfused mouse mTALH, assessed the concentration dependence of blockade of this apical membrane K+-conductive pathway by Ba++ to provide estimates of the magnitudes of the transcellular (Gc) and paracellular (Gs) electrical conductances (millisiemens per square centimeter). These studies also evaluated the effects of luminal hypertonicity produced by urea on the paracellular electrical conductance, the electrical Na+/Cl- permselectivity ratio, and the morphology of in vitro mTALH segments exposed to peritubular antidiuretic hormone (ADH).

PGE2, forskolin, and cholera toxin interactions in rabbit cortical collecting tubule.

To define further the mechanism whereby prostaglandin (PG) E2 inhibits the hydroosmotic response to ADH, we studied the interactions of PGE2 with ADH and two nonhormonal activators of adenylate cyclase, forskolin and cholera toxin, in the isolated perfused rabbit cortical collecting tubule. Forskolin increased hydraulic conductivity (LP) in a dose-dependent fashion and to a degree comparable with ADH-stimulated LP. Forskolin also augmented maximal ADH-stimulated LP, from 135 +/- 15 (SE) to 174 +/- 7 .

Control of NaCl transport in the thick ascending limb.

The mammalian renal thick ascending limb of Henle serves to dilute the urine and supply energy for countercurrent multiplication as a consequence of its ability to absorb NaCl in excess of water. During the past few years our notions of the mechanism of and the factors that control NaCl absorption by the thick ascending limb of Henle have been altered substantially. This review presents evidence for a model of NaCl absorption in the thick ascending limb in which net Cl- absorption is rheogenic and involves a secondary active transport process.

Effects of antidiuretic hormone on cellular conductive pathways in mouse medullary thick ascending limbs of Henle: II. determinants of the ADH-mediated increases in transepithelial voltage and in net Cl-absorption.

Cellular impalements were used in combination with standard transepithelial electrical measurements to evaluate some of the determinants of the spontaneous lumen-positive voltage, Ve, which attends net Cl- absorption, JnetCl, and to assess how ADH might augment both JnetCl and Ve in the mouse medullary thick ascending limb of Henle microperfused in vitro. Substituting luminal 5 mM Ba++ for 5 mM K+ resulted in a tenfold increase in the apical-to-basal membrane resistance ratio, Ra/Rbl, and increasing luminal K+ from 5 to 50 mM in the presence of luminal 10(-4)M furosemide resulted in a 53-mV depolarization of apical membrane voltage, Va. Thus K+ accounted for at least 85% of apical membrane conductance.