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  • Title: Adenosine-induced hyperpolarization of the membrane voltage in rat mesangial cells in primary culture.
    Author: Pavenstädt H, Ruh J, Greger R, Schollmeyer P.
    Journal: Br J Pharmacol; 1994 Sep; 113(1):7-12. PubMed ID: 7529114.
    Abstract:
    1. The effect of adenosine on membrane voltage and ion currents was studied in rat mesangial cells in primary culture. Membrane voltage was measured with the patch clamp technique in the slow- or fast whole cell configuration. The resting membrane voltage of mesangial cells was -48 +/- 0.5 mV. Adenosine (10(-8)-10(-3) M) induced a sustained and concentration-dependent hyperpolarization of membrane voltage (ED50 approximately 6 x 10(-7) M). Adenosine (10(-5) M) hyperpolarized the membrane voltage by 14 +/- 0.5 mV. During the hyperpolarization ion currents were monitored simultaneously. An increase of the outward current by 51 +/- 11% was observed. 2. An increase of the extracellular K+ concentration (from 3.6 to 18.6 M) caused a depolarization of membrane voltage to -34 +/- 2 mV. In the presence of increased K+ the hyperpolarization of membrane voltage induced by adenosine was significantly attenuated by 61 +/- 5%. The K(+)-channel blocker, Ba2+ (5 x 10(-3) M) depolarized membrane voltage to -24 +/- 2 mV. In the presence of Ba2+ the adenosine-induced hyperpolarization was significantly inhibited by 72 +/- 8%. 3. Preincubation of the adenosine antagonist, 8-phenyltheophylline (10(-4) M) significantly inhibited the adenosine (10(-5) M) mediated membrane voltage response by 67 +/- 8%. The adenosine agonists 5-N-ethylcarboxamidoadenosine (NECA), R-(-)N6-(2-phenylisopropyl)adenosine (R-(-)-PIA), S-(+)-N6-(2-phenylisopropyl)adenosine (S-(+)-PIA), N6-[2-(3,5-dimethoxyphenyl)-2-(2-methylphenyl)-ethyl]adenosine (DPMA), and 2-chloroadenosine (2-CA) also hyperpolarized membrane voltage of mesangial cells. The rank order of potency of the agonists at 10-5 M was NECA> adenosine = > R-(-)-PIA = DPMA = 2-CA > S-( + )-PIA.4. Stimulation of cyclic AMP by forskolin induced a concentration-dependent hyperpolarization of membrane voltage (ED50 ~2 x 10-7 M). Application of forskolin (10-5 M) in the presence of adenosine(10-4 M) had no additive hyperpolarizing effect on the membrane voltage.5. Activation of protein kinase C by phorbol 12,13 dibutyrate (PDBu) induced a sustained depolarization of membrane voltage (ED50~ 5 x 10-9 M). In the presence of PDBu, adenosine (10-5 M) still hyperpolarized membrane voltage of mesangial cells.6. The data indicate that adenosine activates K+-conductance via an A2 receptor in mesangial cells; the activation of the K+-conductance, which is probably mediated by cyclic AMP led to a hyperpolarization of membrane voltage.
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