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Isolated muscle fibers from the motor legs of the crab Trichodactilus dilocarcinus were submitted to strong hyperpolarizing currents of varied intensities which produced tension during the current pulse. Threshold for tension was obtained with intensities of about 0.2 x 10(-5) A, changing E(m) to ca. -150 mV (starting from a resting potential ofca. -80 mV). At the closure of the anodic square pulse, a second phase of tension usually appeared superimposed upon the one obtained during hyperpolarization. The first phase of tension increased with the increase of Ca(++) concentration in the bath. Sr(++) produced the same type of mechanical output as Ca(++). When added to the normal Ca(++) concentration, Ba(++) and Mn(++) in low concentrations (up to 21.5 mM) also increased the tension of this phase, but at higher concentrations they blocked both phases while Mg(++) did not alter the tension. Of all the divalent cations employed, only Sr(++) is capable of developing tension as a substitute for Ca(++) in the external media. Procaine administered in a dosage (5 x 10(-3) W/V)which would suppress the contracture due to caffeine (10 mM), did not modify the tension developed during the hyperpolarization. The preceding data indicate that the Ca(++) required for tension during hyperpolarization comes from sites which would differ from those usually postulated for tension due to depolarization in the muscle fibers of other crustaceans (American crayfish). Furthermore, the external source of Ca(++) appears to be one mainly implicated in the induction of tension due to inward current pulses.

1. Membrane potentials were recorded from single rods in the isolated retina of Bufo marinus while the ionic composition of the extracellular medium was rapidly changed. Substitution of 2 mM aspartate(-) for Cl(-) produced a prompt depolarization of horizontal cells, but no modification of either resting potential or response to light in receptor cells. This implies that feed-back from horizontal cells to receptor cells was not active.2. During substitution of choline(+) or Li(+) for Na(+), and during isosmotic substitution of sucrose for NaCl, the resting potential of receptor cells became more negative and responses to light were abolished. During exposure to K(+)-free medium, the resting potential became slightly more negative and the responses to light became larger and developed small after-depolarizations. Exposure to [K(+)](out) of four times normal resulted in permanent diminution of response magnitude and permanent change of response waveshape. Removal of Mg(2+), four times normal [Mg(2+)](out) or substitution of methylsulphate(-) for Cl(-) had no effect on resting potential or responses to light. With the exception of the small effects seen with altered [K(+)](out) these results are consistent with the receptor potential being generated by a light-induced decrease of membrane conductance to Na(+).3. Exposure to decreased [Ca(2+)](out) caused both a depolarization of the receptor membrane in the dark and an increase in the magnitude of the maximal response that could be evoked by a test stimulus. The magnitude of the increase in response equalled the magnitude of the depolarization. Exposure to increased [Ca(2+)](out) or steady background light caused both a steady hyperpolarization and a decrease in the magnitude of the maximal response that could be evoked by a test stimulus. For steady hyperpolarizations greater than 3.5 mV, whether caused by elevated [Ca(2+)](out) or steady background light, the decrease in response magnitude exceeded the magnitude of the hyperpolarization. These results imply that externally applied Ca(2+) ions mimic the effects of steady background lights, but the applied Ca(2+) ions must do more than merely decrease membrane conductance to Na(+).

1. A new simple method is described for quantitating the adhesiveness of circulating polymorphonuclear leucocytes, or granulocytes, to the walls of blood vessels. The cheek pouch of anaesthetized hamsters or a small part of the mesentery of anaesthetized mice were prepared for continuous microscopic observation of selected venules. Those granulocytes which moved sufficiently slowly to be individually visible were counted for 1 or 2 min periods as they rolled past a selected point on one side of a vessel. The velocity distribution of these cells was determined by analysing films. Films were used also to measure mean blood flow velocity in the venules by observing embolizing platelet thrombi induced by the iontophoretic application of adenosine diphosphate. Emigration of granulocytes into the tissues was quantitated by enumerating them in standard areas of stained histological sections.2. In control experiments with hamster cheek pouch venules, the rolling granulocyte count usually passed through a maximum shortly after the preparation was set up and then fell to a low constant value. In mouse mesentery venules the count remained at a low approximately constant value from the beginning for at least 3 hr.3. The mean velocity of blood flow in the venules was between 900 and 200 mu/sec. All rolling granulocytes moved much more slowly; in hamster cheek pouch venules the mean velocity was about 20 mu/sec and in mouse mesentery venules about 10 mu/sec. Around these means the velocity distribution of individual cells was narrow.4. Rolling of granulocytes was abolished by superfusing ethylenediamine tetra-acetate (EDTA, 0.1 M) suggesting that the phenomenon depends on calcium or magnesium ions.5. Agents were applied locally to the observed venules. Human serum albumin, trypsin or histamine in high concentrations did not affect the rolling granulocyte count.6. The rolling granulocyte count was increased during the application of Hammarsten casein or Escherichia coli culture filtrate which are chemotactic to granulocytes in vitro. These agents did not cause alterations in mean blood flow velocity in the observed venules which might have accounted for the effect on the rolling granulocyte counts. When E. coli culture filtrate was perfused through mouse intestine the increase in rolling granulocyte count in the draining venous blood was proportional to the logarithm of the concentration of filtrate.7. The rolling granulocyte count was also increased by the local application of plasma globulin permeability factor or lymph node permeability factor.8. Granulocyte counts in standard histological sections showed no significant increases in control preparations but considerable increases following the application of Hammarsten casein.

1. Single barnacle muscle fibres from Megabalanus psittacus (Darwin) were internally perfused and the effects of various internal and external solutions on voltage clamp currents were examined.2. The usual internal solution was 180 mM-K(+) aspartate (osmotic pressure adjusted to 1000 m-osmole by adding sucrose). Fibres perfused with this solution gave an average resting potential of -55 +/- 5 mV (all potentials are referred to the external solutions as ground). Further increase in internal K concentration depolarized the fibres.3. With membrane current control the total capacitance, referred to apparent membrane surface area, was 21.2 +/- 2.4 muF/cm(2).4. Under normal conditions, with demonstrably good longitudinal space clamp control, voltage clamp currents associated with certain depolarizing pulses showed oscillations. These oscillations were reduced in frequency and magnitude by lowering the temperature from 20 to 10 degrees C, by eliminating the inward currents with external Ca-free saline or by reducing the outward currents with internal tetraethylammonium (TEA) or replacement of internal K by Cs.5. With a Na- and Ca-free, 60 mM-MgCl(2) solution outside depolarizing voltage clamp pulses produced only outward currents. On repolarization the current tail reversed direction at about -70 mV for pulses of less than 50 msec duration. For longer pulses this reversal potential was less negative, suggesting an accumulation of external, or depletion of internal K.6. Both the size of the outward currents and the rate at which they reached their maximum value increased with temperature. The activation energy for the rate constant was about 63 kJ/mole.7. Fibres bathed in Na- and Mg-free, 60 mM-CaCl(2) saline were excitable. After replacement of the internal K(+) with Cs(+) or adding 60 mM-TEA to the internal solution only sustained inward currents were recorded with depolarization.8. Sustained inward currents could be reduced by external application of 5 mM-LaCl(3). Tetrodotoxin was not effective even at a concentration of 1000 nM.9. The rate at which these inward currents reached a maximum value increased with increase in temperature of the bathing solution with an activation energy of the order of 42 kJ/mole.10. The reversal potential of the inward currents changed with the level of internal Ca ions. For a fibre perfused without ethyleneglycol-bis (beta-aminoethyl ether) N,N'-tetraacetic acid (EGTA) this reversal potential was 175 mV (internal free Ca 5 x 10(-7)M), and was 196 mV for a fibre perfused with 20 mM-Tris EGTA (internal free Ca 0.26 x 10(-8)M).11. We propose an electrical equivalent circuit to account for most of the observed electrical properties of barnacle muscle fibres. In this model the Ca and the K system are located at different anatomical places and they interact through a series resistance.

1. Removal of calcium from Carassius either with chelating agents or by adaptation to deionized water increased sodium influx through the gills twofold.2. Treatment of fish with chelating agents increased sodium outflux, whereas adaptation to deionized water reduced sodium outflux. The effects on outflux do not appear to result from calcium removal.3. Addition of calcium (2-20 mM) reversed the effects on influx of chelating agents and of adaptation to deionized water.4. Magnesium (2-10 mM) was without significant effect on sodium influx, but increased the outflow in fish adapted to deionized water.

Skin electrodes are the most convenient reference electrodes for clinical measurements of electrical potential differences (pd) across the epithelium of the alimentary tract but the presence of an electrical charge on normal skin introduces an error. In the present study, by comparison with results obtained using subcutaneous and intravenous electrodes, it was shown that an intradermal injection of saline abolished the skin potential differences. This simple method, therefore, allows skin electrodes to be used to measure the true transepithelial potential differences of gut mucosa. The method was applied to investigate the effect on the rectal potential difference of altering the composition of the luminal solutions. Changes in the cations (sodium, potassium, magnesium) showed that sodium was the most important cationic determinant of the potential difference, especially when sodium absorption was stimulated by giving mineralocorticoids. Changes in the anions (chloride, iodide, bromide, nitrate, bicarbonate, sulphate, phosphate, citrate, and acetate) indicated that the molecular size of the anion rather than its chemical nature was the significant factor and suggested that the ions had to cross a barrier relatively impermeable to anions of radius greater than 3.5 to 4 A degrees . Changes in osmolality and glucose concentration were without effect.