The effect of leakage on micro-electrode measurements of intracellular sodium activity in crab muscle fibres.

The effect of lowering extracellular Na (Nao) on the intracellular Na activity has been measured in single muscle fibres from the crab Carcinus maenas using Na+-sensitive glass micro-electrodes. Measurements have been made with recessed-tip micro-electrodes inserted radially into intact fibres, and with axial electrodes in cannulated fibres. Reducing Nao to one-tenth normal caused local contractions in intact fibres. The apparent steady-state internal Na (Nai) and the fall in Nai when Nao was reduced were found to vary considerably not only between different fibres but also when measured with different electrodes in the same fibre. The steady-state Nai, and the extent and rate of its decrease when Nao was reduced, could be reduced by pushing the Na+-sensitive electrode deeper into the fibre. Cannulated fibres generally had higher internal Na activities than intact fibres, but at comparable levels of Nai the rate of fall recorded from cannulated fibres when Nao was reduced was much slower than with intact fibres. In both intact and cannulated fibres the decrease in Nai was reduced by ouabain. The level of Nai recorded in cannulated fibres was sensitive to depolarization. Depolarizations from -50 to -30 mV resulted in a rise in Nai while further depolarization to 0 mV resulted in a fall in Nai. We conclude that both our results and those of Vaughan-Jones (1977) on undissected fibres are contaminated by leakage into the fibre round the micro-electrode. The true internal Na activity is probably much lower than previously reported.