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M P Ramírez-Ponce

Publications and source records attributed to M P Ramírez-Ponce.

7 recordsLinked to original sources

Human adipose cells have voltage-dependent potassium currents.

The whole-cell patch-clamp method was used to study the membrane electrical properties of human adipocyte cells obtained by differentiating from precursors of human abdominal and mammary tissues. All differentiated cells exhibited outward currents with sigmoidal activation kinetics. The outward currents showed activation thresholds between -20 to -30 mV and slow inactivation. The ionic channels underlying the macroscopic current were highly selective for K(+). Their selectivity was for typical K(+) channels with relative permeabilities of K(+)>NH4+>Cs(+)>Na(+). No evidence of any other type of voltage-gated channel was found. The potassium currents ( I(KV)) were blocked reversibly by tetraethylammonium and barium. The IC(50) value and Hill coefficient of tetraethylammonium inhibition of I(KV) were 0.56 m M and 1.17 respectively. These results demonstrate that human adipose cells have voltage-dependent potassium currents.

Adipose Tissue↗

Insulin increases the density of potassium channels in white adipocytes: possible role in adipogenesis.

We studied the potassium currents in white adipocytes obtained by culturing preadipocytes from rat epididymal tissue, both with insulin (WA(i)) and without insulin (WA(o)), in order to test the role of insulin in the development of voltage-gated potassium channels (K(v)) during adipogenesis. Occasionally, very small potassium currents (I(K,V)) were present in preadipocytes; however these currents were measured in all differentiated cells (adipocytes). WA(i) exhibited greater macroscopic potassium currents than WA(o) with no apparent differences in kinetics or voltage dependence. The current density (pA/ micro m(2)) calculated in WA(i) was higher than in WA(o). Currents were blocked by millimolar concentrations of tetrethylamonium (TEA). The effect of insulin on adipogenesis, both with and without TEA, was analysed. Four days without insulin and three days with insulin were necessary to increase the total number of cells in culture by 2.5-fold. Insulin increased the number of differentiated cells by 73.5%. Cell proliferation and differentiation were inhibited by TEA. Proliferation was affected only by high concentration of TEA. Inhibition of differentiation was dose dependent, with the concentration necessary for half-block similar to the IC(50) values to block potassium channels. These results suggest that insulin increases the density of K(v) and that these channels may be necessary for the normal growth of white adipocytes in culture.

Adipocytes↗

Noradrenaline modulates the electrical activity of white adipocytes by a cAMP-dependent mechanism.

We studied the effect of the external addition of noradrenaline (NA) on the electrical activity of white adipocytes from rat epididymal tissue. NA modified the active voltage response recorded by the current clamp technique, and decreased the macroscopic membrane potassium currents measured by a whole-cell configuration patch-clamp technique in cells obtained by culturing differentiating preadipocytes. To analyse if cAMP is implicated in the effect of this hormone on potassium conductances, the changes in electrical activity caused by an increase in the intracellular concentration of cAMP were studied using intracellular recording. The addition of forskolin, or NN-dibutyryl cAMP plus 3-isobutyl-1-methylxanthine to the bath reduced active changes in membrane voltage responses to hyperpolarizing and depolarizing pulses. A similar effect was observed when vasoactive intestinal peptide was added to the superfusion chamber. These results suggest that NA could modulate K+ conductances in white adipocytes, mediated by cAMP.

1-Methyl-3-isobutylxanthine↗

Voltage-dependent potassium channels in white adipocytes.

Macroscopic membrane currents of white adipocytes were studied using the whole-cell variant of the patch-clamp technique. These cells were obtained by differentiating preadipocytes fro rat epididymal tissue. In all differentiated cells outward currents exhibiting sigmoidal activation kinetics were observed with amplitudes ranging from 0.5 to 6 nA. The outward current showed activation thresholds at approximately -25 to -35 mV. The ion channels underlying the macroscopic current were highly selective for K+. Their selectivity was typical for K+ channels, with relative permeabilities of K+ > NH4+ > Cs+, Na+. The potassium current was blocked by tetraethylammonium, 4-aminopyridine, barium and cobalt. These results demonstrate the existence of voltage dependent potassium channels in mature white adipose cells.

4-Aminopyridine↗

Effects of noradrenaline and insulin on electrical activity in white adipose tissue of rat.

An active change in membrane voltage responses to hyperpolarizing pulses has been identified by intracellular recording on an in vitro preparation of white adipose tissue. This change was characterized by a slow return to baseline at the offset of the pulses. Amplitude and duration of the slow return to baseline were dependent on extracellular K+ concentration, and were diminished by external application of Ba2+. Such properties suggest that this electrical response can be mainly due to activation of transient K+ conductances. The effects that noradrenaline and insulin have over the slow return to baseline have been also studied. While external addition of noradrenaline decreased amplitude and duration of this electrical response, insulin produced the opposite effect. These results suggest that noradrenaline and insulin could modulate K+ conductances in white adipocytes.

Adipose Tissue↗

Electrical activity in white adipose tissue of rat.

Intracellular recording of white adipocytes was performed in an in vitro preparation. Resting potential, input resistance and membrane time constant averaged: -34 +/- 9 mV, 295 +/- 161 M omega, and 58 +/- 19 ms respectively (mean +/- SD, n = 32). Intracellular injection of positive and negative square current pulses elicited membrane voltage responses, characterized by a rectification of the voltage change evoked by positive pulses, and a slow return to baseline at the offset of hyperpolarizing pulses. The amplitude and duration of the slow return to resting potential was dependent on membrane potential, pulse duration, and extracellular K+ concentration. This response was depressed when external Ca2+ was replaced by Co2+, and by external application of 4-aminopyridine. These results indicate that white adipocytes can generate membrane voltage responses which may mostly be a consequence of the activity of ionic channels. The properties of the slow return to baseline suggest that it may be due to a transient K+ current.

4-Aminopyridine↗

Photooxidase system of Rhodospirillum rubrum III. The role of rhodoquinone and ubiquinone in the activity of preparations of chromatophores and photoreaction centers.

The role of rhodoquinone and ubiquinone in the oxygen photoreducing (photooxidase) activity of Rhodospirillum rubrum was investigated. The sole addition of purified rhodoquinone restored photooxidase activity in isolated chromatophores which had been extracted with organic solvents and which were apparently free of secondary acceptor ubiquinone. Rhodoquinone also enhanced photooxidase activity in photoreaction center preparations from which secondary ubiquinone seemed to have been removed. Those results suggest that rhodoquinone accepts electrons directly from primary ubiquinone during chromatophore photooxidase activity. In contrast, rhodoquinone does not participate in the basal activity of photoreaction center preparations, which seems to result from the autooxidation of both primary and secondary ubiquinone.

Bacterial Chromatophores↗