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Biomedical subjects

E Schlatter

Publications and source records attributed to E Schlatter.

At least 91 records · Page 5Linked to original sources
PubMed1985

Diphenylamine-2-carboxylate, a blocker of the Cl(-)-conductive pathway in Cl(-)-transporting epithelia.

The present study examines the effects of diphenylamine-2-carboxylate (DPC) in Cl(-)-transporting epithelia. This substance blocks reversibly the Cl(-)-conductance present under normal circumstances in the basolateral membrane of the thick ascending limb of the loop of Henle (TAL) and in the apical membrane of shark rectal gland tubules (RGT). This leads to a reduction in active NaCl reabsorption (TAL) and NaCl secretion (RGT) respectively, as measured by the equivalent short circuit current. The cells hyperpolarize as the membrane voltage drifts from the control value (some compromise between the chemical potential of Cl- and K+) towards the chemical potential of K+. The resistance of the basolateral (TAL) or apical membrane (RGT) increases and this leads to a moderate increase in transepithelial resistance. In addition, the Cl(-)-concentration step induced membrane voltage changes, which can be produced under control conditions, disappear in the presence of the blocker. Finally, experiments in excised membrane patches indicate that this substance inhibits the single current events of individual Cl(-)-channels.

Animals↗
PubMed1985

Cl- -channels in the apical cell membrane of the rectal gland "induced" by cAMP.

Isolated rectal gland tubules (n approximately equal to 1000) of dogfish (Squalus Acanthias) were perfused in vitro. Individual channels in the apical and basolateral cell membrane were recorded with the patch clamp method. K+-channels were present in excised membrane patches of the basolateral membrane in stimulated (dbcAMP + forskolin + adenosine) and in nonstimulated state. Cl- -channels were found only in patches of the apical cell membrane when the tubule was stimulated. Cell attached recordings and simultaneous transepithelial PD measurements were obtained while the segment was stimulated. It is shown that concomitant with the increase in lumen negative PD "silent" membrane patches of the apical cell membrane suddenly develop Cl- -channel activity. It is concluded that stimulation of rectal gland tubules "activates" Cl- -channels in the apical cell membrane.

Animals↗
PubMed1985

cAMP increases the basolateral Cl- -conductance in the isolated perfused medullary thick ascending limb of Henle's loop of the mouse.

The effect of cAMP on transepithelial and transmembrane potential differences and resistances was examined in isolated in vitro perfused mouse medullary thick ascending limbs of Henle's loop (mTAL). The effects of furosemide and barium were tested. Stimulation of NaCl transport by ADH 10(-9) + dbcAMP 4 X 10(-4) + forskolin 10(-6) mol X l-1 (paired experiments) resulted in: a) an increase in transepithelial potential difference, referenced to the grounded bath, from +6.7 +/- 0.3 mV to +12.0 +/- 0.4 mV (n = 47); b) a decrease in transepithelial resistance from 25 +/- 1 omega cm2 to 20 +/- 1 omega cm2 (n = 47); c) a depolarization of the basolateral membrane by 12 mV and of the apical membrane by 7 mV (n = 36); d) a decrease in the fractional resistance of the basolateral membrane from 0.27 +/- 0.05 to 0.15 +/- 0.06 (n = 12). Furosemide (10(-4) mol X l-1) abolished the active transepithelial transport potential and hyperpolarized the basolateral membrane potential to values which were similar in both control and cAMP treated mTAL segments. Barium increased the transepithelial resistance and depolarized PDb1 to similar values in both functional states. An increase in the fractional conductance of the basolateral membrane was also seen, if, prior to the cAMP treatment, the luminal Na+2Cl-K+ cotransport was inhibited by furosemide. Thus, we propose that stimulation of active NaCl reabsorption in the mTAL segment of the mouse by ADH, mediated via cAMP, increases primarily the basolateral chloride conductance.

Animals↗
PubMed1984

Potassium activity in cells of isolated perfused cortical thick ascending limbs of rabbit kidney.

The Na+2Cl-K+ cotransporter in the apical membrane of the cortical thick ascending limb of the Henle's loop (cTAL) of rabbit nephron utilizes the electrochemical gradient for Na+ to transport K+ and Cl- against an unfavorable electrochemical gradient from lumen to cell interior. In the present study attempts are made to measure intracellular K+ activity (a cell K+) under control conditions and after inhibition of the cotransport system by furosemide (50 X 10(-6) mol X l-1). 70 cTAL segments of 55 rabbits were perfused in vitro. Conventional Ling-Gerard and K+-selective microelectrodes were used to measure the PD across the basolateral membrane (PDbl) as well as the PD sensed by the single barrelled K+-selective electrode (PDK+). PDbl was -64 +/- 1 (n = 65) mV and PDK+ + 15 +/- 1 (n = 32) mV under control conditions. The positive PDK+ value, significantly different from zero, indicates that a cell K+ is higher than predicted for passive distribution. The estimate for a cell K+ obtained from PDbl and PDK+ was 113 +/- 8 mmol X l-1. Furosemide lead to the previously reported hyperpolarization of PDbl by 17 +/- 4 (n = 13) mV and to a reduction of PDK+ from 15 +/- 1 to 5 +/- 1 (n = 20) mV. The a cell K+, obtained from this set of data, was 117 +/- 9 mmol X l-1, and was not different from the control value. The present data indicate that a cell K+ is significantly above Nernst equilibrium under control conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). II. Effects of inhibitors.

Rectal gland tubule (RGT) segments of the spiny dogfish (Squalus acanthias) were perfused in vitro. The effects of inhibitors of known mode of action on transepithelial PD (PDte resistance (Rte), the PD across the basolateral membrane (PDbl), the fractional resistance of this membrane (FRbl), and intracellular activities of NA+, Cl-, K+ (apha cell) were examined. Furosemide (5 x 10(-4) mol x 1(-1)) reduced PDte from -12 +/- 0.7 to -2.3 +/- 0.2 mV (n = 63), hyperpolarized PDbl from -71 +/- 1.3 to -79 +/- 0.9 mV (n = 59), FRbl decreased from 0.2 +/- 0.03 to 0.13 +/- 0.01 (n = 21), alpha cell cl- fell from 38 +/- 4 to 11 +/- 2 mmol x 1(-1) (n = 21), alpha cell Na+ fell from 37 +/- 4 to 17 +/- 2 mmol x 1(-1) (n = 12) and alpha cell K+ was constant [113 +/- 14 vs. 117 +/- 15 mmol x 1(-1) (n = 6)]. Furosemide exerted its effects within some 20-40s. Its action was completely reversible. Analysis of the time courses revealed that the furosemide induced initial fall in alpha cell cl- was approximately twice as rapid when compared to that of alpha cell Na+. Ba2+ 0.5 mmol x 1(-1) (bath) reduced PDte from -7.1 +/- 1.2 to -4.1 +/- 0.6 mV (n = 24), increased Rte from 18 +/- 2 to 22 +/- 2.5, omega cm2 (n = 14). PDbl depolarized from -75 +/- 2 to -48 +/- 2 mV (n = 42), FRbl increased from 0.2 +/- 0.02 to 0.34 +/- 0.04 (n = 14) and alpha cell K+ increased from 143 +/-28 to 188 +/- mmol x 1(-1) (n = 4). Ouabain (50 x 10(-6) mol x 1(-1), bath) reduced PDte from -12 +/-2 to -3 +/- 0.5 mV (n = 9), Rte increased from 18 +/- 3 to 21 +/- 3 omega cm2 (n = 5). PDbl depolarized from -67 +/- 4 to -26 + 3 mV (n = 14), FRbl increased from 0.23 +/- 0.04 to 0.45 +/- 0.05 (n = 6), alpha cell K+ fell only slightly from 135 +/- 15 to 112 +/- 30 mmol x 1(-1) (n = 4), but alpha cell cl- increased from 35 +/- 12 to 111 +/- 37 mmol x 1(-1) (n = 3). These effects of ouabain were slow when compared to those exerted by furosemide or Ba2+. The ouabain effects on PDte and PDbl were completely prevented if furosemide was applied first.(ABSTRACT TRUNCATED AT 400 WORDS)

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗
PubMed1984

Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). III. Effects of stimulation of secretion by cyclic AMP.

Segments of rectal gland tubules (RGT) the spiny dogfish (Squalus acanthias) were perfused in vitro to study the cellular mechanism by which NaCl secretion is stimulated. Transepithelial PD (PDte), transepithelial resistance (Rte), the PD across the basolateral membrane (PDbl), the fractional resistance of the lumen membrane (FR1), and the cellular activities for Cl-, Na+, and K+ (alpha cell x) were measured. In series 1 the effects of stimulation (S) (dbcAMP 10(-4, adenosine 10(-4), and forskolin 10(-6) mol x 1(-1) on these parameters were recorded and compared to nonstimulated state (NS). PDte increased from -1.9 +/- 0.2 mV to -11.0 +/- 0.9 mV (n = 51). PDbl depolarized from -86 +/- 1 to -74 +/- 1.4 mV (n =52), Rte fell from 29 +/- 2.8 to 21 +/- 2 omega cm2 (n = 23), and FR1 fell from 0.96 +/- 0.005 to 0.79 +/- 0.04 (n = 9). alpha cell K+ was constant (123 +/- 13 versus 128 +/- 17 mmol x 1(-1) (n = 6), but alpha cell cl- fell significantly from 48 +/- 4 to 41 +/- 3 mmol x 1(-1) (n = 7). alpha cell Na+ increased from 11 +/- 2.1 to 29.5 +/- 6.6 mmol x 1(-1) (n = 4). In series 2 the conductivity properties were examined by rapid K+, and Cl- concentration steps on the basolateral and luminal cell side respectively in NS and S states. In NS-segments reduction of bath K+ led to a hyperpolarization of PDbl with a mean slope of 28 +/- 1.3 mV/decade (n = 9) (as compared to 19 mV/decade for S-state).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Substrate utilization in the isolated perfused cortical thick ascending limb of rabbit nephron.

Isolated segments of cortical thick ascending limbs (cTAL) of rabbit kidney were perfused in vitro and the equivalent short circuit current (Isc) was measured. In a first series all substrates were removed on either side. Isc fell rapidly to 50 +/- 12% after 3 min and to 27 +/- 6% (n = 5) after 10 min. This indicates that in cTAL segments Isc is strictly dependent on the presence of substrates. In series two it was tested what substrates can be utilized by the cTAL segment, and from which epithelial side [bath (b) or lumen (1)] the substrates are taken up. From the 1-side only butyrate (10 mmol X 1(-1) sustained the Isc at 95 +/- 2% (n = 7). All other tested substrates (10 mmol X 1(-1): pyruvate, acetate, beta-OH-butyrate, D-glucose, and L-lactate lead to a marked decline in Isc. From the b-side several substrates (5--10 mmol X 1(-1) sustained the Isc: D-glucose, D-mannose, butyrate, beta-OH-butyrate, acetoacetate, L-lactate, acetate and pyruvate. Other compounds (1--10 mmol X 1(-1): citrate, alpha-ketoglutarate, succinate, glutamate, glutamine, propionate, caprylate and oleate did not sustain Isc. In the third series the mechanism of substrate utilization from the basolateral cell side was studied. It was shown that the Isc is a saturable function of the D-glucose, L-lactate, acetate, pyruvate or beta-OH-butyrate concentration with apparent Km's between 0.05--1.0 mmol X 1(-1). Several known inhibitors of sugar and of anion transport were tested at the bath side: phlorrhizin was without effect. Phloretin (500 mumol X 1(-1) inhibited Isc by 96%, yet its effect was not dependent on the presence of substrates on the b-side since inhibition occurred also if the b-perfusate contained no substrate and Isc was driven by luminal butyrate. Also SITS (5 mmol X 1(-1) exerted only a small inhibitory effect which was not specific since it was also observed with luminal butyrate. alpha-Cyano-m-OH-cinnamate (10 mmol X 1(-1) inhibited the Isc specifically when L-lactate was the bath substrate. Probenecid (1 mmol X 1(-1) had a similar yet less marked inhibitory effect. The D-glucose uptake from the b-side was specifically inhibited by cytochalasin B at 5 X 10(-6) mol X 1(-1). We conclude that the cTAL segment of the rabbit utilizes D-glucose and/or small anions such as pyruvate or L-lactate or acetate to energize salt reabsorption.(ABSTRACT TRUNCATED AT 400 WORDS)

Absorption↗
PubMed1984

Mechanism of NaCl secretion in the rectal gland of spiny dogfish (Squalus acanthias). I. Experiments in isolated in vitro perfused rectal gland tubules.

Rectal gland tubules (RGT) of spiny dogfish were dissected and perfused in vitro. Transepithelial PD (PDte), resistance (Rte), the PD across the basolateral membrane (PDbl) and intracellular chloride and potassium activities (a cell Cl-, a cell K+) were measured. In a first series, 67 RGT segments were perfused with symmetric shark "Ringers" solution. The bath perfusate contained in addition db-cAMP 10(-4), forskolin 10(-6), and adenosine 10(-4) mol X 1(-1). PDte was -11 +/- 1 (n = 67) mV lumen negative, Rte 27 +/- 2 (n = 47) omega cm2, PDbl -75 +/- 0.4 (n = 260) mV. a cell K+ and a cell Cl- were 109 +/- 22 (n = 4) and 38 +/- 4 (n = 36) mmol X 1(-1) respectively. These data indicate that Cl-secretion across the RGT must be an uphill transport process, whereas secretion of Na+ could be driven by the lumen negative PDte. Intracellular K+ is 14 mV above equilibrium with respect to the basolateral membrane PD and Cl- is 23 mV above equilibrium across the apical membrane. In series 2, the conductivity properties of the apical and basolateral membrane as well as that of the paracellular pathway were examined in concentration step experiments. Decrease of the basolateral K+ concentration led to a rapid hyperpolarization of PDbl with a mean slope of 19 mV per decade of K+ concentration change. Addition of 0.5 mmol X 1(-1) Ba2+ to the bath solution lead to a marked depolarization and abolished the response to K+ concentration steps. In the lumen a Cl- concentration downward step led to a depolarization of the lumen membrane; resulting in a mean slope of 18 mV per decade of Cl- concentration change. When dilution potentials were generated across the epithelium, the polarity indicated that the paracellular pathway is cation selective. In series 3 the equivalent short circuit current (Isc = PDte/Rte) was determined as a function of symmetrical changes in Na+ concentration, with Cl- held at 276 mmol X 1(-1), and as a function of symmetrical changes in Cl- concentration, with Na+ held at 278 mmol X 1(-1). Isc was a saturable function of Na+ concentration (Hill coefficient 0.9 +/- 0.1, K1/2 4.4 mmol X 1(-1), n = 7) and also a saturable function of Cl- concentration (Hill coefficient 2.0 +/- 0.1, K1/2 75 mmol X 1(-1), n = 11).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

Cellular mechanism of the action of loop diuretics on the thick ascending limb of Henle's loop.

During the passed few years the cellular mechanisms responsible for the NaCl reabsorption in the thick ascending limb of the Henle loop of mammalian nephron and of the early distal tubule of amphibian kidney have been extensively studied. From these studies a new type of secondarily active transport mechanism, i.e. the Na+--2Cl- --K+ symport has emerged. Meanwhile it has been recognized that this system is also present in many other epithelia. All these epithelia share in common that they are sensitive to the so called loop diuretics. The present article summarizes our current knowledge of how the loop diuretics, by reversible interaction with the above cotransport system, inhibit the NaCl reabsorption in the thick ascending limb of Henle's loop. It is shown that these drugs transfer the thick ascending limb cell to a state in which not only transepithelial NaCl reabsorption ceases but in which also very little energy is consumed since then K+ and Cl- "relax" to passive distribution across both cell membranes and Na+ entry into the cell is blocked.

Absorption↗
PubMed1983

Effect of "high ceiling" diuretics on active salt transport in the cortical thick ascending limb of Henle's loop of rabbit kidney. Correlation of chemical structure and inhibitory potency.

The group of "high ceiling" diuretics consists of a variety of chemically different potent diuretic and saluretic substances. Appart from a few exemptions direct evidence for an action of these substances in the thick ascending limb of the loop of Henle (TAL) is still lacking. For furosemide, we have reported recently that it inhibits most likely the Na+-2 Cl--K+ cotransport system present in the lumen membrane of the TAL. The present study tests: 1. whether other "high ceiling" diuretics have a similar site and mode of action, and 2. how modifications of the furosemide molecule alter the inhibitory potency. Isolated cortical TAL (cTAL) segments (n = 185) of rabbit kidneys were perfused in vitro. The equivalent short circuit current (Isc = transepithelial PD/transepithelial resistance), as a measure of active salt transport was correlated to the dose of 64 substances. Several diuretics, such as 2-aminomethyl-4-(1,1-dimethyl-ethyl)-6-iodophenol hydrochloride (MK 447), hydrochlorothiazide, muzolimine, etozoline, tizolimide, amiloride, and triamterene were ineffective both from the lumen and basolateral side at concentrations as high as 10(-4) - 10(-3) mol X 1(-1). The phenoxyacetic acids ethacrynic acid, indacrinone (MK 196), and to less an extend tienilic acid were inhibitory active. They differed from furosemide in one or more of the following criteria: delayed onset, incomplete reversibility, stronger action from the bath, different slope of the dose response curve. Similarly, 1-ozolinone acted stronger from the bath. In contrast, the diuretics of the furosemide type and related compounds (bumetanide and piretanide) showed rapid onset and complete reversibility of inhibition. These substances acted stronger from the lumen. The individual positions in the benzyl ring of the diuretics were differently affected by substitutions, leading to parallel shifts in the dose response curves with halfmaximal inhibition at concentrations ranging between 8 X 10(-8) to greater than 10(-4) mol X 1(-1). For these substances the calculated Hill coefficients were close to unity: 0.96 +/- 0.05. We conclude that the so called "high ceiling" or "loop" diuretics consist of at least 3 groups: 1. drugs that do not interfere with the active salt transport in the cTAL segment, 2. drugs that interfere by so far not characterised mechanisms, and 3. drugs of the furosemide type which inhibit the Na+-2 Cl--K+ cotransport system in the lumen membrane of the cTAL segment.

Animals↗
PubMed1983

Chloride activity in cells of isolated perfused cortical thick ascending limbs of rabbit kidney.

Rabbit cortical thick ascending limb segments were perfused in vitro, and intracellular Cl- activity was estimated in three types of experiments using conventional and chloride selective microelectrodes. In series 1 Ringer like solutions were present on the two epithelial sides. In series 2 limen Cl- was replaced by gluconate, and in series 3 furosemide, 10-20 . 10(-6) mol . 1-1, was added to the lumen perfusate. It was found that under control conditions intracellular Cl- activity, as estimated from the difference of the reading of the conventional (n = 53) and ion selective electrodes (n = 118) was 26 +/- 1 mmol . 1-1. Thi value is approximately three times higher than expected for passive distribution of Cl-. After removal of lumen Cl- (series 2) intracellular Cl- activity fell to 9 mmol . 1(-1) which is only some 4 mmol . 1(-1) above passive distribution. We argue that these 4 mmol . 1(-1) reflect mainly the interference with the Cl- electrode by other anions, such as phosphate. The above estimates for intracellular Cl- activity, have to be diminished by these 4 mmol . 1(-1), and, thus, are close to 22 mmol . 1(-1). In series 3 a rapid and reversible fall in intracellular Cl- from 23 to 7 mmol . 1(-1) was observed. We conclude that the Cl- activity in cTAL cells is clearly above equilibrium under control conditions and that it falls rapidly to values close to equilibrium when Cl- reabsorption is blocked by either removing lumen Cl- or by blocking the Cl- entry via the Na+-2 Cl--K+-carrier with furosemide.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1982

Renal handling of homologous and heterologous insulin in the isolated perfused rat kidney.

1. Renal handling of pig- and rat-insulin was studied in the isolated perfused rat kidney. 2. Metabolic clearance rates of both pig- and rat-insulin exceeded GFR. 3. Peritubular uptake of pig-insulin accounted for 13% of rat-insulin for 31% of the total metabolic clearance. 4. The nonfiltering kidney does not remove insulin from the peritubular circulation. 5. Metabolic clearance rates of pig- and rat-insulin are directly related to GFR. 6. The filtration process seems to be necessary for the uptake of insulin at the peritubular site.

Animals↗
PubMed1982

A cytotoxin of Pseudomonas aeruginosa acts directly on the cortical thick ascending limb of Henle's loop of rabbit kidney.

The present study examines directly the effect of a cytotoxin of Pseudomonas aeruginosa on the in vitro perfused rabbit cortical thick ascending limb of the loop of Henle (cTAL). 25 cTAL segments were perfused at high rate. The open circuit transepithelial electrical PD (PDte) and the specific electrical transepithelial resistance (Rt) were recorded continuously. From PDte/Rt the equivalent short circuit current (Isc) was calculated. The Isc was 214 +/- 30 mu A.cm-2 under control conditions, and decreased significantly to 74 +/- 34 mu A.cm-2 60 s after the addition of toxin (2 mg.1(-1)) to the lumen perfusate. Microscopic observation and photographs taken at that time clearly indicated swelling of the cTAL cells. Thereafter inhibition of active transport proceeded further, Rt fell progressively, and cells started to desquamate from the basement membrane. This effect of the toxin was dose dependent, and was half maximal at approximately 1.2 mg.1(-1). From the bath side the effect was less marked and higher doses of toxin had to be used (half maximal effect at 5 mg.1(-1)). We conclude that this toxin of Pseudomonas aeruginosa exerts its toxic effect on the cTAL segment by increasing primarily the permeability of the lumen membrane.

Animals↗
PubMed1982

Renal handling of uremic middle molecules. a study with the isolated perfused rat kidney.

Little is known of the normal renal handling of potential uremic toxins in the middle molecular weight (MMW) range (300-2,000 daltons). In this study the isolated rat kidney has been used as a model to investigate renal clearance and possible renal metabolic effects on uremic MMW species. Isolated rat kidneys were perfused with hemofiltrate from chronic renal failure patients containing 5% bovine albumin. Middle molecules (MM) were isolated from perfusates and urines by high-speed gel filtration (peak 7) and further separated by gradient elution chromatography to give 7-8 definable uremic MMW species (7a, b, c, etc.). Species 7b, d, e and f were tubularly reabsorbed by 15, 25, 23 and 34%, respectively, as indicated by simultaneous inulin clearances. 7a was freely filtered with negligible reabsorption. 1 MM (7c1) was found to be secreted by up to 220% at low perfusate concentrations, secretion being masked at high perfusate levels. No MMs were catabolized by the kidney, although one species (7c2) appeared to be synthesized. The results suggest that uremic MMs have differing biological or physicochemical activities as indicated by the selectivity of renal handling.

Animals↗
PubMed1981

Presence of luminal K+, a prerequisite for active NaCl transport in the cortical thick ascending limb of Henle's loop of rabbit kidney.

Previous data from our laboratory have shown that active transport in the cortical thick ascending limb of Henle's loop (cTAL), as measured by the short circuit current (Lsc, microA X cm-2), requires the presence of Na+ and Cl-. The data were compatible with the model of secondarily active Cl- reabsorption involving the cotransport of Na+ and Cl- across the luminal membrane. The data suggested, furthermore, that 1 Na+ and 2 Cl- interact with the luminal carrier. In the present study it was tested whether this reabsorptive mechanism also requires the presence of luminal K+. Isolated cTAL segments (n = 40) were perfused at high flow rates with a modified Ringer's solution. Removal of K+ from the lumen reduced Isc significantly from 215 to 133 microA X cm-2. Addition of Ba2+ (10(-3) mol X 1(-1)) which blocks the K+ conductance of the luminal, membrane, to the K+-containing lumen perfusate decreased Isc significantly from 234 to 141 microA X cm-2. Combination of both manoeuvres: perfusion with a K+-free and Ba2+-containing solution almost abolished Isc from a control of 237 to 56 microA X cm-2. The results are compatible with the view that in rabbit cTAL the luminal carrier interacts with all 3 ions, possibly 1 Na+, 2 Cl-, and 1 K+. K+ recycles across the luminal membrane through its conductive pathway.

Animals↗