SUMMARY1. Histological evidence suggests that the force deficit associated with eccentric contraction-induced muscle injury is due to structural damage to contractile elements within the muscle fibre. Alternatively, the force deficit could be explained by an inability to activate the contractile proteins. It was the objective of this study to investigate the latter possibility.2. Mouse soleus muscles were isolated, placed in an oxygenated Krebs-Ringer buffer at 37°C, and baseline measurements were made. The muscle then performed one of three contraction protocols: (1) twenty eccentric (n = 10 muscles); (2) ten eccentric (n = 12); or (3) twenty isometric (n = 10) contractions. At the end of the injury protocol, measurements were made during performance of a passive stretch, twitch and tetanus. Next, force was recorded during exposure of the muscle to buffer containing 50 mm caffeine.3. Decrements in maximal isometric tetanic force (P0) observed for muscles in the twenty eccentric, ten eccentric, and twenty isometric contraction protocols were 42-6 + 4-2, 20-0 + 2-3 and 3.9 + 2-4 %, respectively. However, the caffeine-elicited forces in muscles from the three protocols were not different when corrected for initial differences in P0 (64X9 + 1X3, 64-2 + 2-1 and 68-9 + 2-5 % of pre-injury P0). The peak caffeine-elicited force was 118-4+8-6% of post-injury Po for the muscles in the twenty eccentric contraction protocol, which was significantly different from that observed for the other protocols (71-8-80-2 % post-injury P0). These findings indicate that the force deficit in this muscle injury model results from a failure of the excitation process at some step prior to calcium (Ca2+) release by the sarcoplasmic reticulum.4. In an attempt to locate the site of failure, intracellular measurements were made in injured muscles to test whether injury to the sarcolemma might have resulted in a shift of the resting membrane potential of the muscle fibre. However, microelectrode measurements of resting membrane potential for muscles in the twenty eccentric contraction protocol (-74-4 + 0-6 mV) were not different from muscles in the twenty isometric contraction protocol (-73-4+ 1-0 mV). These data suggest that membrane resting conductances were normal and are compatible with MS 1641 G. L. WARREN AND OTHERS the idea that the ability of the injured fibres to conduct action potentials was probably not impaired.

Activity-dependent variations in extracellular potassium concentration in the central nervous system may be regulated, in part, by potassium spatial buffering currents in glial cells. The role of spatial buffering in the retina was assessed by measuring light-evoked potassium changes in amphibian eyecups. The amplitude of potassium increases in the vitreous humor was reduced to approximately 10 percent by 50 micromolar barium, while potassium increases in the inner plexiform layer were largely unchanged. The decrease in the vitreal potassium response was accurately simulated with a numerical model of potassium current flow through Müller cells, the principal glial cells of the retina. Barium also substantially increased the input resistance of Müller cells and blocked the Müller cell-generated M-wave, indicating that barium blocks the potassium channels of Müller cells. Thus, after a light-evoked potassium increase within the retina, there is a substantial transfer of potassium from the retina to the vitreous humor by potassium current flow through Müller cells.

The retina of the frog was superfused with a Ringer solution containing impermeant "probe" cations and anions. Light-evoked concentration changes in these probe ions were measured in the subretinal space (SRS) with ion-selective microelectrodes. A decrease in probe ion concentration was found, and several observations suggest that this is caused by a light-evoked expansion of the SRS. The probe ion decrease was not seen in the isolated retina; thus, the pigment epithelial (PE) cells are important for its generation. Pharmacological studies suggest that K+ channels in the PE cells are important--perhaps the PE cells shrink in response to the light-evoked decrease in SRS [K+]. The light-evoked decrease of SRS volume may be important in the understanding of SRS solute concentrations, retina-PE adhesivity, photoreceptor-PE cell interactions, and the interphotoreceptor matrix.

1. The actions of two pharmacological agents, barium ions (Ba2+) and picrotoxin (PTX), were examined on components of the electroretinogram (ERG) in frog retina. Depth profiles of light-evoked field potentials were recorded, and current source densities (CSDs) were computed from these. 2. Ba2+ abolished the M-wave, slow PIII, and the c-wave, but only decreased b-wave amplitude down to approximately 65% of control amplitude. 3. Ba2+ abolished a slow current sink in the inner plexiform layer (IPL) and the source at the inner limiting membrane (ILM). This IPL sink/ILM source appears to generate the M-wave. 4. Ba2+ decreased the current sink at the outer plexiform layer (OPL) to approximately 70% of control amplitude, and it increased an IPL source. This Ba(2+)-resistant OPL sink/IPL source appears to generate a significant portion of the b-wave. The Ba(2+)-sensitive portion of the b-wave might be generated by Müller cells. 5. PTX enhanced retinal field potentials, particularly the M-wave in the proximal retina. This enhanced M-wave was shown to originate from an enhanced IPL sink/ILM source. 6. Our results suggest that the M-wave originates from Müller cells, through the spatial buffering of the light-evoked increase in [K+]o of the proximal retina. A portion of the b-wave may also originate from Müller cells, but a stronger direct contribution from depolarizing bipolar cells is suggested.

Measurements of absolute transretinal resistance and of the relative resistance of the various retinal layers were obtained in the frog. The resistance of clamped sections of isolated retina was 66 omega . cm2, which results in an average resistivity (rho) between inner and outer limiting membranes of 5,050 omega . cm. In eyecups, relative resistances (obtained by passing constant currents across the retina) were assigned to specific layers of the retina with the aid of physiological criteria (e.g., depths of light-evoked field potentials, changes in extracellular K+ concentration, base-line noise level, and resistance). These relative resistances were then converted to absolute values, a calculation feasible because the region between inner and outer limiting membranes, which has the same structure in both isolated and eyecup retinas, could be specified during experiments. Resistivities (in omega . cm) for the retinal layers include 1) subretinal space, 970; 2) inner and outer nuclear layers, 6,800; and 3) inner plexiform layer, 1,750. The ganglion cell and optic nerve fiber layers were too thin to resolve individually, but rho of the two layers combined was 7,900. The outer plexiform layer was also too thin to reliably resolve, but its rho is likely the same as the inner plexiform layer. The extracellular space volume fraction (alpha) of the retinal layers was estimated from these rho s, and the following values were obtained: 1) subretinal space, 0.12; 2) outer and inner nuclear layers, 0.03; 3) inner and outer plexiform layers, 0.11; and 4) ganglion cell and optic nerve fiber layers, 0.02. The decreased rho and increased alpha of the inner plexiform layer and the subretinal space, compared with that of the nuclear layers, are expected from their anatomy. A consideration of these inhomogeneities is required in analyses of field potentials and of changes in extracellular ionic concentrations.