A new inhibitory pathway in the jellyfish Polyorchis penicillatus

Contact of food with the manubrial lips in the genus Polyorchis A. Agassiz, 1862 evokes trains of electrical impulses (E potentials) that propagate to the margin. E potentials are also produced by food stimuli at the margin and tentacle bases. E potentials are shown to be associated with inhibitory postsynaptic potentials (ipsps) in the swimming motor neurons and contribute to the arrest of swimming during feeding. The conduction pathway for E potentials is a nerve plexus located in the endodermal walls of the stomach and radial and ring canals. We have explored the conducting properties of the system; the conduction velocity varies with stimulus frequency but is about 15 cm/s when stimuli are more than 50 s apart. Neurites belonging to the E system run around the margin adjacent to the inner nerve ring, where the swimming pacemaker neurons are located. We suggest that they may make inhibitory synapses on to the swimming motor neurons, but this has yet to be demonstrated anatomically. The reversal potential for ipsps, recorded intracellularly with potassium acetate micropipettes, was estimated to be about –69 mV. Swimming inhibition mediated by this endodermal pathway is distinct from that observed during protective “crumpling” behaviour and that associated with contractions of the radial muscles seen during feeding, though it may accompany the latter.

Calcium currents from jellyfish striated muscle cells: preservation of phenotype, characterisation of currents and channel localisation

SUMMARY When striated muscle cells of the jellyfish Polyorchis penicillatus were dissociated at 30°C they retained their in vivo morphology and the integrity of ionic currents. This contrasted with cells dissociated at room temperature that rarely expressed any inward currents. Whole-cell, patch-clamp recordings from dissociated muscle cells revealed that the inward component of the total ionic current consisted of only one calcium current. This calcium current activated at –70 mV, peaked at –30 mV, and inactivated within 5 ms. In comparison with barium and strontium ions, calcium ions were the preferred current carriers. Calcium channels can be blocked by dihydropyridines and nickel ions at micromolar levels. Several properties of this current are reminiscent of T-type calcium currents. Localisation of this channel using the fluorescent channel blocker fDHP and the fluorescent dye RH414 indicated that myofibres had a higher density of these channels than the somata.

Modulation of Jellyfish Potassium Channels by External Potassium Ions

The amplitude of an A-like potassium current ( I Kfast) in identified cultured motor neurons isolated from the jellyfish Polyorchis penicillatus was found to be strongly modulated by extracellular potassium ([K+]out). When expressed in Xenopus oocytes, two jellyfish Shaker-like genes, jShak1 and jShak2, coding for potassium channels, exhibited similar modulation by [K+]out over a range of concentrations from 0 to 100 mM. jShak2-encoded channels also showed a decreased rate of inactivation and an increased rate of recovery from inactivation at high [K+]out. Using site-directed mutagenesis we show that inactivation of jShak2 can be ascribed to an unusual combination of a weak “implicit” N-type inactivation mechanism and a strong, fast, potassium-sensitive C-type mechanism. Interaction between the two forms of inactivation is responsible for the potassium dependence of cumulative inactivation. Inactivation of jShak1 was determined primarily by a strong “ball and chain” mechanism similar to fruit fly Shaker channels. Experiments using fast perfusion of outside-out patches with jShak2 channels were used to establish that the effects of [K+]out on the peak current amplitude and inactivation were due to processes occurring at either different sites located at the external channel mouth with different retention times for potassium ions, or at the same site(s) where retention time is determined by state-dependent conformations of the channel protein. The possible physiological implications of potassium sensitivity of high-threshold potassium A-like currents is discussed.

Voltage sensing in jellyfish Shaker K+ channels.

The S4 segment of the jellyfish (Polyorchis penicillatus) Shaker channel jShak1 contains only six positively charged motifs. All other Shaker channels, including the jellyfish Shaker channel jShak2, have seven charges in this segment. Despite their charge differences, both these jellyfish channels produce currents with activation and inactivation curves shifted by approximately +40 mV relative to other Shaker currents. Adding charge without changing segment length by mutating the N-terminal side of jShak1 S4 does not have a pronounced effect on channel activation properties. Adding the positively charged motif RIF on the N-terminal side of K294 (the homologue of K374 in Drosophila Shaker, which is a structurally critical residue) produced a large positive shift in both activation and inactivation without altering the slope of the activation curve of the channel. When IFR was added to the other side of K294, there was a small negative shift in activation and fast inactivation of the channel was prevented. Our results demonstrate that K294 divides the S4 segment into functionally different regions and that the voltage threshold for activation and inactivation of the channel is not determined by the total charge on S4.