Quantitative analysis of single muscle fibre action potentials recorded at known distances

In vivo records of single fibre action potentials (SFAPs) have always been obtained at unknown distance from the active muscle fibre. A new experimental method has been developed enabling the derivation of the recording distance in animal experiments. A single fibre is stimulated with an intracellular micropipette electrode. The same electrode is used thereafter for labelling with an auto-fluorescent dye, Lucifer Yellow. In this method there is no use of chemical fixation. The tissue structure is kept as well as possible. In cross-sections the fluorescent fibre is seen and its position is quantitized with respect to the tip of one or more recording wire electrodes. Morphometric data, such as the recording distance and the fibre cross-sectional area, are used for the interpretation of parameters of the SFAPs (peak-peak amplitude, time between the first positive and negative peaks). The present results show that within 300 microns recording distance is not as dominant for the SFAP shape as expected. The method offers also a direct check of the relation between the muscle fibre; diameter and the conduction velocity of the action potential. In the present small set of data there is no simple linear relationship.