Electrical properties of Lupinus angustifolius L. stem. III. RC model, time constant, latency and threshold charge

The changes of potential in the stem of <i>Lupinus</i> were characterized on the basis of the strength-duration relation and of electrotonic potentials. It was found that the stimulated stem behaves like an electrical RC circuit. The time constants of electrotonic potential rise and decay were determined. A simple electrical model characterizing the passive electrical properties of the <i>Lupinus</i> stem is suggested. The values of resistance and capacitance of the <i>Lupinus</i> stem were determined on the basis of the RC circuit. The resistance-capacitance properties of the stem tissues serve as basis to gain a better knowledge of the parametres describing excitation, such as the strength-duration relation or latency. These properties in the stem of <i>Lupinus</i> are of the same nature as those in nerves or muscles. The values of the threshold charge of the order of 10-6 C were calculated. It is suggested that the regularities occurring here may be connected with accommodation and processes regulating the resting potential of cells.

Electrical conduction within the cerebrovasculature of stroke-prone spontaneously hypertensive rats

Alterations in electrical conductivity between smooth muscle cells (SMCs) can alter the spread and effectiveness of electromechanical SMC contraction. We attempted to determine whether alterations in pressure-dependent constriction (PDC) occurring in relation to stroke development within the middle cerebral arteries (MCAs) of Wistar-Kyoto stroke-prone hypertensive rats (SHRsp) were associated with changes in electrical conductivity between the SMCs. Current was injected into nonpressurized MCAs, using a suction electrode. The conducting distance along the length of the MCA where the amplitude of the membrane potential deflection (electrotonic potential) produced by current injection declined to 1/e (length constant) was used to measure conductivity. PDC to a 100 mmHg pressure step was measured with a pressure myograph. A loss of PDC in the MCAs of SHRsp preceded stroke development. Heptanol (4 mM), a gap junction communication inhibitor, reversibly inhibited conductivity and PDC in the MCA of prestroke SHRsp. The ability of heptanol to reversibly inhibit PDC was likely not related to it's ability to alter electrical conduction. The length constant of electrical conduction in the MCAs was about 0.75 mm and didn't differ between MCA sampled from pre- versus post-stroke SHRsp or Sprague-Dawley rats. It was concluded that alterations in electrical conductivity along the MCA could modify the spread of PDC, but such changes do not contribute to the loss of PDC within the MCA of poststroke SHRsp.Key words: membrane potential, electrotonic potential, middle cerebral arteries, myogenic response, gap junction, stroke-prone hypertensive rats.

Mechanism of block by ZD 7288 of the hyperpolarization-activated inward rectifying current in guinea pig substantia nigra neurons in vitro

1. The effects of the novel bradycardic agent 4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyrimidinium chloride (ZD 7288) (Zeneca) were investigated on the hyperpolarization-activated cationic current (Ih) in guinea pig substantia nigra pars compacta neurons in vitro, using a single-microelectrode current-clamp/voltage-clamp technique. 2. Under current-clamp conditions, injection of large negative current pulses (0.1-0.5 nA, 400 ms) evoked a slow depolarizing "sag" in the electrotonic potential due to activation of the slow inward (anomalous) rectifier. In voltage-clamp recordings, hyperpolarizing voltage steps from a holding potential of -60 mV (close to resting potential) elicited slow inward current relaxations with kinetic properties similar to those seen for other neuronal Ihs. 3. ZD 7288 (10-100 microM) produced a consistent abolition of the electrotonic potential sag with no effect on membrane potential or spike properties. Under voltage clamp, Ih amplitude was clearly reduced in a time- and concentration-dependent manner (apparent half-maximum blocking concentration = 2 microM); full block of Ih was typically achieved after 10-15 min of exposure to 50 microM ZD 7288, with no significant recovery observed after 1 h of washing. 4. A similar (although more rapid) block of Ih was seen after application of 3-5 mM Cs+ (partially reversible after 30 min of washing). 5. Partial block of Ih by 10 microM ZD 7288 was accompanied by a reduction in the maximum amplitude of the Ih activation curve, a small negative shift in its position on the voltage axis, and a linearization of the steady-state current-voltage relationship. The estimated Ih reversal potential, however, remained unaffected. 6. In 10 microM ZD 7288, the time course of Ih activation and deactivation was significantly slowed (within the range of -70 to -120 mV for the activation time constant and -70 to -90 mV for the inactivation time constant). 7. Blockade of Ih by ZD 7288 or Cs+ was independent of prior Ih activation (i.e., non-use dependent). 8. Intracellular loading with ZD 7288 also abolished the sag in the electrotonic voltage response and Ih relaxations, suggesting an intracellular site of action. By contrast, intracellular Cs+ had no effect on Ih properties. 9. Block of Ih by ZD 7288 (but not Cs+) was relieved by prolonged cell hyperpolarization, manifested as a slowly developing (half-time approximately 20 s) inward current at a holding potential of -100 mV. 10. We propose that ZD 7288, when applied externally, may behave as a "lipophilic" quaternary cation, capable of passing into the cell interior to block Ih channels in their closed state; this compound may thus prove a useful research tool, in place of Cs+, for studying the properties and significance of Ih currents in controlling neuronal function.

Intercellular Communication in an Insect Endocrine Gland

1. The parenchymal cells of the corpora allata (CA) of the cockroach Diploptera punctata are coupled through low-resistance intercellular pathways. 2. Extensive dye coupling of cells of CA from fourth instar and adult female cockroaches was revealed following iontophoretic injection of Lucifer Yellow. 3. Freeze-etch electron microscopy revealed numerous gap-junction-like particles in CA cell plasma membranes. 4. The spread of ionic current from cell to cell was demonstrated by injecting current pulses into one cell and recording electrotonic potentials from other cells. The amplitude of electrotonic potentials elicited by negative current injection varied inversely with the rate of juvenile hormone biosynthesis by the CA. 5. The ‘length constant‘ (distance at which an electrotonic potential in CA cells decays to 37% of its magnitude at the site of current injection) could not be measured accurately, but was found to be much greater than the length of the CA. 6. Forskolin- and IBMX-induced elevation of intracellular cAMP increased the electrotonic potential but reduced juvenile hormone release of day 8 CA.

Mechanisms underlying pattern generation in lobster stomatogastric ganglion as determined by selective inactivation of identified neurons. III. Synaptic connections of electrically coupled pyloric neurons

1. The pyloric dilator (PD) and anterior burster (AB) neurons in the pyloric system of the lobster stomatogastric ganglion are electrically coupled and synchronously active. We have used the lucifer yellow photoinactivation technique to separate the connections made by the PD motor neurons from those made by the AB interneuron. 2. Photoinactivation of either the two PD neurons or the single AB neuron allowed us to separate the compound inhibitory postsynaptic potentials (IPSPs) in the lateral pyloric (LP) and pyloric (PY) motor neurons resulting from synchronous PD and AB activity into AB-evoked and PD-evoked components. These IPSPs have different time courses, reversal potentials, ion selectivities, and pharmacological properties. 3. Photoinactivation and membrane-potential manipulations indicated that a readily observable IPSP recorded in the AB neuron and correlated with action potentials in the LP neuron is actually an electrotonic potential due to an LP-evoked IPSP in the PD neurons. 4. Selective inactivation of either the two PD neurons or the AB neuron revealed that the IPSP recorded in the ventricular dilator (VD) motor neuron is due solely to AB-released transmitter. 5. The electrical coupling potentials measurable between the AB, PD, and VD neuron somata are due to direct electrical coupling between all of these neurons. 6. Circuit analysis and transmitter identification may be complicated by electrical coupling. We suggest that the presence of electrical coupling between nonidentical neurons may provide a new mechanism that allows changes in synaptic characteristics among neurons within a "hard-wired" circuit.

Electrical coupling and uncoupling of exocrine acinar cells.

The electrical communication network in the mouse pancreatic acinar tissue has been investigated using simultaneous intracellular recording with two separate microelectrodes and direct microscopical control of the localizations of the microelectrode tips. All cells within one acinus were electrically coupled, and the coupling coefficient (the electrotonic potential change in a cell neighboring to the cell into which current is injected [V2] divided by the electrotonic potential change in the cell of current injection [V1]) between two cells near each other (less than 50 micron) was always close to 1. Cells farther apart (50-100 micron) were, in some cases, coupled; in other cases, there was no coupling at all. Coupling coefficients varied between 0 and 1. There was rarely electrical coupling over distances of more than 110 micron. Using microiontophoretic acetylcholine (ACh) application, it was possible to evoke almost complete electrical uncoupling of two previously coupled pancreatic or lacrimal acinar cells from different acini or within one acinus. The effects were fully and quickly reversible. While the ACh-evoked uncoupling in the pancreas was associated with membrane depolarization, ACh caused hyperpolarization in the lacrimal acinar cells. The uncoupling was associated with a very marked reduction in electrical time constant, indicating a reduction in input capacitance (effective surface cell membrane area). The concentrations of stimulants needed to evoke reduction in pancreatic cell-to-cell coupling were 1 micron for ACh, 0.14 nM for caerulein, and 3 nM for bombesin. These concentrations are smaller than those required to evoke maximal enzyme secretion.

Biophysical Effects of Adrenaline on the Smooth Muscle of the Rabbit Common Carotid Artery

Effects of adrenaline on the smooth muscle of the rabbit common carotid artery were studied by the partitional chamber method. The experiments on excitation-contraction coupling were carried out in isotonic Krebs solution; the other experiments were carried out in hypertonic Krebs solution. Adrenaline (10-7 g/ml) caused rhythmical electrical and mechanical activity of arterial strips in isotonic Krebs solution. By addition of adrenaline (10-5 g/ml), the membrane was depolarized by about 10 mv and the amplitude of the electrotonic potential was decreased by 40–50% of the control in hypertonic Krebs solution. Present experimental results suggest that the depolarization of the membrane and the decrease of the amplitude of the electrotonic potential in the artery are due to the increase of Na and Cl conductance. Contraction appeared in all preparations exposed to 10-8 g/ml adrenaline; at that concentration membrane potential and membrane resistance showed little or no change.