Voltage-dependent K+ currents in rat cardiac dorsal root ganglion neurons.

We have assessed the expression and kinetics of voltage-gated K(+) currents in cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of a fluorescent tracer into the pericardial sack. Ninety-nine neurons were labelled: 24% small (diameter<30 microm), 66% medium-sized (diameter 30 microm>.48 microm) and 10% large (>48 microm) neurons. Current recordings were performed in small and medium-sized neurons. The kinetic and pharmacological properties of K(+) currents recorded in these two groups of neurons were identical and the results obtained from these neurons were pooled. Three types of K(+) currents were identified:a) I(As), slowly activating and slowly time-dependently inactivating current, with V(1/2) of activation -18 mV and current density at +30 mV equal to 164 pA/pF, V(1/2) of inactivation at -84 mV. b) I(Af) current, fast activating and fast time-dependently inactivating current, with V(1/2) of activation at two mV and current density at +30 mV equal to 180 pA/pF, V(1/2) of inactivation at -26 mV. At resting membrane potential I(As) was inactivated, whilst I(Af), available for activation. The I(As) currents recovered faster from inactivation than I(Af) current. 4-Aminopiridyne (4-AP) (10 mM) and tetraethylammonium (TEA) (100 mM) produced 98% and 92% reductions of I(Af) current, respectively and 27% and 66% of I(As) current, respectively. c) The I(K) current that did not inactivate over time. Its V(1/2) of activation was -11 mV and its current density equaled 67 pA/pF. This current was inhibited by 95% (100 mM) TEA, whilst 4-AP (10 mM) produced its 23% reduction. All three K(+) current components (I(As), I(Af) and I(K)) were present in every small and medium-sized cardiac DRG neuron. We suggest that at hyperpolarized membrane potentials the fast reactivating I(As) current limits the action potential firing rate of cardiac DRG neurons. At depolarised membrane potentials the I(Af) K(+) current, the reactivation of which is very slow, does not oppose the firing rate of cardiac DRG neurons.

Voltage-dependent Ca2+ currents in rat cardiac dorsal root ganglion neurons.

This study presents the kinetic and pharmacological properties of voltage-gated Ca(2+) currents in anatomically defined cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of fluorescent tracer Fast Blue into the pericardial sack. There were three distinct groups of neurons with respect to cell size: small (27% of total; cell capacitance <30 pF), medium (65% of total; capacitance 30-80 pF) and large neurons (8% of total; capacitance >80 pF). The properties of Ca(2+) currents were tested in small and medium-sized neurons. In large neurons currents could not be adequately controlled and were not analysed. Ca(2+) currents did not completely inactivate during 100 ms depolarising voltage steps. The activation thresholds in small (-36.9+/-1.3 mV) and medium (-39.0+/-1.3 mV) size neurons were similar. Current densities were 105.8 pA/pF in small and 97.4 pA/pF in large neurons and also did not differ. The kinetic properties of activation and inactivation did not differ between small and medium-sized cardiac DRG neurons. At membrane potentials between -50 and -60 mV (the expected resting membrane potential in these neurons) 55 to 70% of Ca(2+) currents in small and medium-sized neurons were available for activation. Both, small and medium-sized neurons expressed similar proportions of L (7.5%), N (25%) and P/Q (36%) type Ca(2+) currents. We conclude that small and medium-sized cardiac DRG neurons are homogeneous with respect to the expression and properties of voltage-gated Ca(2+) currents. Voltage-gated Ca(2+) currents probably play an important role in action potential generation in cardiac DRG neurons due to their availability for activation at resting membrane potential, their high density and voltage threshold close to the threshold for voltage-gated Na(+) currents.

Properties of Na+ currents in putative submandibular and cardiac sympathetic postganglionic neurones.

This study was performed to compare the kinetic properties of Na+ currents in putative salivary and cardiac postganglionic sympathetic neurones isolated from the superior cervical and stellate ganglia, respectively. Neurones were labelled with a fluorescent tracer-Fast Blue, injected into the submandibular gland (in the case of salivary neurones) and into the pericardial cavity or left ventricular wall (in the case of cardiac neurones). Voltage-dependent Na+ current was then isolated and recorded from labelled cells. The major findings of this study were: (1) Peak Na+ current was larger in salivary than in cardiac neurones (5.7 nA vs. 2.4 nA; for 30 mM Na+ in extra- and 15 mM in the intracellular solution). (2) The somata of salivary neurones were twice as large as those of cardiac neurones, as indicated by the values of their membrane capacitance (36 pF vs. 18 pF). (3) There was a greater Na+ current density (169 pA/pF vs. 128 pA/pF) in salivary than in cardiac neurones. (4) Recovery from inactivation was faster in salivary neurones with 90% recovery time being 93 ms for salivary and 144 ms in cardiac neurones. (5) Half-activation times were voltage-dependent and consistently longer for salivary than for cardiac neurones. (6) Remaining parameters, such as current threshold, maximum current voltage and kinetics of steady-state inactivation did not significantly differ in salivary compared to cardiac neurones.

Stellate neurones innervating the rat heart express N, L and P/Q calcium channels.

The aim of the study was to investigate the kinetic properties and identify the subtypes of Ca2+ currents in the cardiac postganglionic sympathetic neurones of rats. Neurones were labelled with a fluorescent tracer--Fast-Blue, injected into the pericardial cavity. Voltage-dependent Ca2+ currents were recorded from dispersed stellate ganglion cells that showed Fast Blue labelling. Only high threshold voltage-dependent Ca2+ currents were found in the somata of cardiac sympathetic neurones. Their maximum amplitude, mean cell capacitance and current density were respectively: 0.67 nA, 19.3 pF and 36.4 pA/pF (n = 21). The maximum Ca2+ conductance was 51.3 nS (n = 14). Half activation voltage equalled +11.0 mV and the slope factor for conductance 11.1 (n = 14). As tested with a 10 s pre-pulse, the Ca2+ current began to inactivate at -80 mV. Half inactivation voltage and slope factor for steady-state inactivation were -36.6 mV and 14.1 (n = 9), respectively. Saturating concentration of L channel blocker (nifedipine), N channel blocker (omega-conotoxin-GVIA), P/Q channel blocker (omega-Agatoxin-IVA) and N/P/Q channel blocker (omega-conotoxin-MVIIC) reduced the total Ca2+ current by 26.8% (n = 7), 57.1% (n = 12), 25.9% (n = 6) and 69.4% (n = 6), respectively. These results show that the somata of cardiac postganglionic cardiac sympathetic neurones contain significant populations of N, L and P/Q high threshold Ca2+ channels.

Distribution of compound potentials recorded from the surface of isolated stellate ganglia following stimulation of postganglionic branches, in rats and cats.

The experiments were performed on 9 cat and 18 rat isolated stellate ganglia. Rats and cats were anesthetized with alpha-glucochloralose or urethane, respectively. The ganglia, isolated with their branches, were transferred to a recording chamber and constantly superfused with artificial extracellular fluid bubbled with 95% O2 and 5% CO2. Branches of the ganglion were one by one placed in suction electrodes and stimulated. Antidromic evoked potentials were systematically recorded from numerous points on the ganglion surface. The area under the curve of the negative wave of each recorded potential was considered proportional to the number of neurons located in the vicinity of the recording electrode, projecting to the stimulated nerve. We have found that: (1) cardiac sympathetic neurons are located in the lower, caudal half of the ganglia; (2) vertebral sympathetic neurons occupy the cranial, upper half of the ganglia; (3) neurons with axons in the ansae are positioned near the point of exit of the respective ansa from the ganglion; (4) localization of neurons projecting to the same branches is very similar on both sides--right and left; (5) this localization is also similar in rats compared to cats.