An electrophysiological method for measuring the potassium permeability of the nerve perineurium.
An electrophysiological method is described for measuring the potassium permeability (PK) of the perineurium of the sciatic nerve of the frog. The method is based on the principle of grease-gap recording, in which an insulating compartment separates two surface recording electrodes. The sciatic nerves of frogs Rana temporaria and R. pipiens were isolated and mounted across a five compartment chamber, with Vaseline grease seals on the partitions between compartments. Compartments #1, #2 and #5 contained frog Ringer solution, #4 was filled with Vaseline and formed the grease gap, and #3 was the test compartment in which solutions could be changed. The nerve was stimulated via platinum electrodes in compartments #1 and #2, and DC potentials and compound action potentials (CAP) were recorded between Ag/AgCl electrodes connected through Ringer-agar bridges to compartments #3 and #5. In nerves with undamaged perineurium, changing from normal Ringer to high [K+] Ringer (100 mM, KCl replacing NaCl) for 2 min caused negligible change in DC potential or CAP, indicating that raised [K+] was not reaching the axon surface, and hence that the perineurium was exerting a diffusional restriction on K+ entry. In nerves damaged by stretching or drying, K+ pulses caused a depolarising change in DC potential (delta DC), and corresponding decline in CAP amplitude, consistent with a leaky perineurium allowing K+ entry and axonal depolarisation. Ringer made hypertonic by the addition of 2.5 M sucrose or 5 M NaCl caused increased perineurial permeability to K+. The method was calibrated by measuring the delta DC in response to raised [K+] in the range 5-100 mM [K+] in desheathed nerves; from this calibration curve relating delta DC to endoneurial [K+] it was possible to calculate the change in endoneurial [K+] occurring in intact preparations. The calculations showed that the undamaged perineurium had a PK of < 6.3 x 10(-7) cm.s-1, similar to the value calculated for in situ nerves using radioisotopic techniques, but less than the value reported for isolated perineurial cylinders. The method gives real-time information on the K+ permeability of the nerve perineurium and its modulation by experimental treatments.
Abbott
N J
NJ
Biomedical Sciences Division, King's College London, UK. nj.abbott@kcl.ac.uk
Mitchell
G
G
Ward
K J
KJ
Abdullah
F
F
Smith
I C
IC
D000818:Animals; D002199:Capillary Permeability / Q000502:physiology*; D004058:Diffusion; D017097:Electric Impedance; D004594:Electrophysiology; D006982:Hypertonic Solutions; D007552:Isotonic Solutions; D008839:Microelectrodes; D008954:Models, Biological; D009995:Osmosis; D010525:Peripheral Nerves / Q000378:metabolism*; D011188:Potassium / Q000493:pharmacokinetics*; D011894:Rana pipiens; D011896:Rana temporaria; D000077331:Ringer's Solution; D012584:Sciatic Nerve / Q000378:metabolism / Q000648:ultrastructure; D012965:Sodium Chloride; D013395:Sucrose
D006982:Hypertonic Solutions; D007552:Isotonic Solutions; D012965:Sodium Chloride; D013395:Sucrose; D000077331:Ringer's Solution; D011188:Potassium