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M Paulmichl

Publications and source records attributed to M Paulmichl.

66 records · Page 4Linked to original sources
PubMed1986

Apparent chloride conductance of subconfluent Madin Darby canine kidney cells.

In incompletely confluent Madin Darby canine kidney (MDCK)-cells continuous measurements of the potential difference across the cell membrane (PD) were made with conventional microelectrodes during rapid changes of extracellular chloride concentration. During control conditions mimicking in vivo situation, PD averages -50.3 +/- 0.7 mV. Reduction of extracellular chloride concentration from 122 mmol/l to 64.5 mmol/l depolarizes the cell membrane by +1.8 +/- 0.2 mV while reduction to 16 mmol/l leads to a transient, variable depolarization followed by a hyperpolarization of the cell membrane by -11.8 +/- 1.4 mV. 1 mmol/l anthracene-9-COOH hyperpolarizes the cell membrane by -10.7 +/- 1.0 mV, and abolishes the effect of altered extracellular chloride concentration (-0.6 +/- 0.5 mV), 1 mumol/l diphenylamine-2-carboxylate hyperpolarizes the cell membrane by -11.7 +/- 1.4 mV. 10 mumol/l furosemide hyperpolarize the cell membrane by -11.4 +/- 1.4 mV. Step increases of extracellular potassium concentration from 5.4 to 20 mmol/l depolarize the cell membrane by +14.9 +/- 1.0 mV in the absence of inhibitors, by +24.2 +/- 1.3 mV in the presence of anthracene-9-COOH and by +28.8 +/- 0.7 mV in the presence of furosemide. 10 mumol/l isoproterenol depolarize the cell membrane by +2.4 +/- 0.3 mV and increase the depolarizing effect of reducing extracellular chloride concentration to 64.5 mmol/l (+2.9 +/- 0.4 mV). 1 mumol/l forskolin depolarizes the cell membrane by +5.8 +/- 1.0 mV. In conclusion, chloride conductance of subconfluent MDCK-cells may be small during control conditions, is apparently decreased by anthracene-9-COOH and reduction of extracellular chloride concentration but is enhanced by isoproterenol.

Animals↗
PubMed1986

Electrical properties of Madin-Darby-canine-kidney cells. Effects of extracellular sodium and calcium.

In incompletely confluent Madin Darby canine kidney cells continuous measurements of the potential difference across the cell membrane (PD) were made with conventional microelectrodes during rapid changes of extracellular sodium and/or calcium concentration. During control conditions PD averages -50.6 +/- 0.7 mV. Reduction of extracellular sodium concentration from 131.8 to 17.8 mmol/l leads to a reversible hyperpolarization of the cell membrane to -65.3 +/- 1.1 mV. This hyperpolarization is not significantly reduced by omission of glucose or presence of amiloride (1 mmol/l) in the perfusates. Instead, 1 mmol/l amiloride depolarizes the cell membrane by +5.2 +/- 0.4 mV. 1 mmol/l barium depolarizes the cell membrane to -31.3 +/- 1.1 mV. Step increases of extracellular potassium concentration from 5.4 to 10 and 20 mmol/l depolarize the cell membrane by +5.5 +/- 0.5 mV and +16.5 +/- 1.8 mV respectively. In the presence of barium, the depolarizing effect of increasing extracellular potassium concentration and of amiloride is almost abolished. Reduction of extracellular sodium concentration in the presence of barium, however, leads to a transient hyperpolarization of the cell membrane. During this transient hyperpolarization, increasing extracellular potassium concentration depolarizes the cell membrane despite the continued presence of barium. Omission of extracellular calcium (EDTA) depolarizes the cell membrane by +36.7 +/- 3.2 mV. In the absence of extracellular calcium, the hyperpolarizing effect of reduced extracellular sodium concentration is markedly reduced (-4.5 +/- 1.2 mV). 2 mumol/l A23187 in the presence of extracellular calcium hyperpolarizes the cell membrane to -72.5 +/- 0.6 mV.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Effects of epinephrine on electrical properties of Madin-Darby canine kidney cells.

The present study has been performed, to test for the influence of epinephrine on the potential difference across the cell membrane (PD) of Madin-Darby canine kidney (MDCK) cells. Under control conditions, mimicking the in vivo situation, PD averages - 53.3 +/- 0.9 mV (n = 37). Increasing extracellular potassium concentration from 5.4 to 10 and 20 mmol/l depolarizes the cell membrane by +4.3 +/- 0.4 mV (n = 5) and +15.8 +/- 1.2 mV (n = 5), respectively. The application of 1 mumol/l epinephrine leads to sustained hyperpolarization of the cell membrane to -71.5 +/- 0.7 mV (n = 37). In the presence of epinephrine, increasing extracellular potassium concentration from 5.4 to 20 mmol/l depolarizes the cell membrane by +30.6 +/- 0.2 mV (n = 5); 1 mmol/l barium depolarizes the cell membrane by +14.8 +/- 0.7 mV (n = 20) and abolishes the effect of step increases of extracellular potassium concentration from 5.4 to 10 mmol/l. In the presence of barium, epinephrine leads to a transient hyperpolarization by -31.2 +/- 1.2 mV (n = 18). During this transient hyperpolarization, the cell membrane is sensitive to extracellular potassium concentration despite the continued presence of barium; 10 mumol/l verapamil depolarizes the cell membrane to -41.0 +/- 2.6 mV (n = 11). In the presence of verapamil, the hyperpolarizing effect of epinephrine is only transient; 10 mumol/l phentolamine depolarizes the cell membrane by +3.0 +/- 0.6 mV (n = 8). In the presence of phentolamine, the effect of epinephrine is virtually abolished (+0.4 +/- 0.6 mV, n = 8); 1 mumol/l isoproterenol depolarizes the cell membrane by +2.8 +/- 0.8 mV (n = 8).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Electrical properties of Madin-Darby canine kidney cells. Effects of extracellular potassium and bicarbonate.

To gain some insight into electrogenic transport processes across the plasma membrane of Madin-Darby canine kidney (MDCK)-cells, continuous measurements of the potential difference across the plasma measurements of the potential difference across the plasma membrane (PD) were made during step changes of extracellular ion composition as well as application of barium or valinomycin. During control conditions mimicking in vivo extracellular fluid, PD approaches -51.5 +/- 0.8 mV (n = 62). Step increase of extracellular potassium concentration from 5.4 to 10, to 20 or to 35 mmol/l, depolarizes PD by +5.5 +/- 0.8 mV (n = 7), by +15.8 +/- 0.5 mV (n = 64) and by +23.8 +/- 1.2 mV (n = 12), respectively. 1 mmol/l barium depolarizes PD by +19.8 +/- 0.6 mV (n = 38) and abolishes the effect of increasing extracellular potassium from 5.4 to 10 mmol/l but not to 35 mmol/l. Ten mumol/l valinomycin hyperpolarizes PD to -69.3 +/- 2.9 mV (n = 7). In the presence of valinomycin, increase of extracellular potassium from 5.4 to 20 mmol/l depolarizes PD by +31.0 +/- 1.0 mV (n = 7). Ouabain depolarizes PD and reduces the sensitivity of PD to extracellular potassium concentration. Omission of extracellular bicarbonate and carbon dioxide as well as increase of extracellular bicarbonate at constant carbon dioxide lead to a hyperpolarization and enhanced sensitivity of PD to extracellular potassium. In the presence of barium, the effects of omitted bicarbonate and carbon dioxide of MDCK-cells is highly conductive to potassium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Ouabain decreases apparent potassium-conductance in proximal tubules of the amphibian kidney.

According to a previous study from this laboratory, the electrochemical gradient for potassium across the peritubular cell membrane of proximal tubules in the isolated perfused frog kidney increases following the application of ouabain. In order to test, if this phenomenon were due to a decrease of potassium conductance, the effects of ouabain on cell membrane resistances and the sensitivity of the peritubular cell membrane potential difference (PDpt) to step changes of peritubular potassium and bicarbonate concentration were studied. In the absence of ouabain, PDpt averaged -60 +/- 3 mV (n = 25). A step increase of peritubular potassium concentration from 3 to 18 mmol/l (pH 8.07) depolarizes PDpt (delta PDk) by +24 +/- mV (n = 8). An increase of bicarbonate from 20 to 40 mmol/l (pH 8.07) hyperpolarizes PDpt (delta PDb) by -2.8 +/- 0.4 mV (n = 9). The resistance of the luminal and peritubular cell membranes in parallel (Rm) amounts to 45 +/- 9 k omega cm (tubule length) (n = 4) and the voltage divider ratio (VDR) to 1.4 +/- 0.2 (n = 7). The resistance of the cellular cable (cellular core, Rc) approaches 131 +/- 37 M omega/cm (n = 4). Peritubular application of 0.1 mmol/l ouabain leads to a gradual decline of PDpt (t1/2 approx. 30 min), to an increase of Rm, a decrease of delta PDk and an increase of delta PDb. VDR and Rc are not changed significantly. The data point to a functional link between the sodium/potassium ATPase and the potassium conductance of the peritubular cell membrane.

Animals↗
PubMed1984

The influence of intracellular sodium activity on the transport of glucose in proximal tubule of frog kidney.

Inhibition of basolateral Na+/K+ ATPase by ouabain eventually abolishes transport of glucose. The present study was performed to test, if this effect is due to a dissipation of the electrochemical gradient for sodium or due to a regulatory inhibition of sodium-coupled glucose entry across the luminal membrane at increasing intracellular sodium activity. To this end, proximal convoluted tubules of the doubly perfused isolated frog kidney were perfused alternatively with solutions containing either 5 mmol/l glucose or raffinose. The potential difference across the peritubular cell membrane (PDpt) and across the epithelium (PDte) has been recorded with conventional and across the peritubular cell membrane with ion selective microelectrodes (PDpt). In the absence of luminal glucose PDpt is (+/- SEM) -54.0 +/- 2.4 mV, PDte = -1.2 +/- 2.0 mV and PDNapt = -96 +/- 5 mV. The electrochemical gradient for sodium (mu Na+) amounts to 95 mV and intracellular sodium activity to 14 mmol/l (extracellular sodium activity is 74 mmol/l). Luminal application of glucose leads to a rapid depolarisation of PDpt (delta PDpt = 8.6 +/- 0.9 mV and PDNapt (delta PDNapt = 11.1 +/- 3.0 mV) and to hyperpolarisation of PDte (delta PDte = -0.8 +/- 0.2 mV). The peritubular application of ouabain leads to a gradual, reversible and proportional decline of PDpt, PDNapt and mu Na+. Glucose induced delta PDpt and delta PDNapt decrease in parallel to PDpt and PDNapt, resp. In a separate series, the lumped conductance (Gm) of the luminal and basolateral cell membrane has been determined, which amounts to 2.4 +/- 0.3 microS/mm (tubule length). Gm decreases 23 +/- 4%, when PDpt is decreased to half.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Hypotonicity and ethanol modulate BK channel activity and chloride currents in GH4/C1 pituitary tumour cells.

AIM: Description of the effects of hypotonic cell swelling and ethanol on maxi Ca2+-activated K+ channel (BK channel) activity and Cl- channel activity in GH4/C1 pituitary tumour cells. METHODS: Whole cell-, cell attached- and outside-out patch clamp measurements, fluorescence (fluo-3) measurements of intracellular Ca2+ concentration, cell size video monitoring. RESULTS: GH4/C1 pituitary tumour cells respond to both hypotonicity and ethanol with cell swelling which is followed by a regulatory volume decrease (RVD). Tetraethylammonium and 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) induced cell swelling per se and inhibited hypotonicity induced RVD. Ethanol-induced swelling is paralleled by an increase in the intracellular Ca2+ concentration and augmented by DIDS. BK channel activation by hypotonicity and ethanol is demonstrated in patch clamp experiments both in intact cells (cell attached configuration) and a subset of excised membrane patches (outside-out configuration). Cell swelling and addition of ionomycin under isotonic conditions leads to the activation of outwardly rectifying Cl- currents with time dependent activation at positive potentials. CONCLUSIONS: In GH4/C1 cells both hypotonicity and ethanol lead to cell swelling, RVD and to activation of BK channels. The hypotonicity-induced BK channel activation can also be observed in cell free outside-out patches. Hypotonicity, but not ethanol leads to the activation of Cl- channels with features of Ca2+-activated Cl- currents.

3T3 Cells↗
PubMedDate not supplied

The ICln interactome.

The many different functional phenotypes described in mammalian cells can only be explained by an intense interaction of the underlying proteins, substantiated by the fact that the number of independently expressed proteins in living cells seems not to exceed 25 K, a number way too small to explain the >250 K different phenotypes on a one-protein-one-function base. Therefore, the study of the interactome of the different proteins is of utmost importance. Here, we describe the present knowledge of the ICln interactome. ICln is a protein, we cloned and whose function was reported to be as divers as (i) ion permeation, (ii) cytoskeletal organization, and (iii) RNA processing. The role of ICln in these different functional modules can be described best as being a 'connector hub' with 'date hub' function.

Binding Sites↗
PubMedDate not supplied

Cell volume regulation in renal cortical cells.

Both proximal renal tubule cells and cultured Madin-Darby canine kidney (MDCK) cells are capable of regulating their volume in hypotonic media. Regulatory cell volume decrease in proximal straight tubules is impaired by barium, amiloride and acetazolamide and depends on the presence of bicarbonate and of sodium, whereas it is unaffected by complete removal of extracellular chloride. The observations may point to parallel loss of potassium through potassium channels as well as of bicarbonate and sodium via a bicarbonate-sodium cotransport. Alternatively, potassium/hydrogen ion exchange or potassium bicarbonate cotransport could be involved. In MDCK cells, exposure to hypotonic media apparently leads to the activation of an anion channel, while potassium conductance is rather decreased. In both proximal tubules and MDCK cells, volume regulatory decrease is possibly triggered by leucotrienes, which may be released during cell swelling. Cell volume is altered in a variety of conditions even at isotonic extracellular fluid and cell volume-regulatory mechanisms are likely to participate in regulation of renal transepithelial transport.

Animals↗
PubMedDate not supplied

Ion channels in Madin-Darby canine kidney cells.

Ion channels in Madin-Darby canine kidney cells serve transepithelial chloride transport and probably cell volume regulation. Three distinct potassium channels and one anion channel have been revealed by patch clamp studies in Madin-Darby canine kidney cells. The potassium channels are activated by an increase in intracellular calcium activity. A number of hormones activate the potassium channels by an increase in intracellular calcium activity. However, under certain conditions the hormones hyperpolarize the cell membrane without increasing intracellular calcium activity sufficiently to activate the calcium-sensitive potassium channels. Thus, the hormones may activate potassium channels via another, as yet undefined, intracellular mechanism. The anion channel is stimulated by cAMP. Another factor modifying channel activity is cell volume: cell swelling leads probably to subsequent activation of potassium and anion channels. The net result is a variable transient hyperpolarization followed by a sustained depolarization of the cell membrane.

Animals↗