Biomedical subjects

L C Rome

Publications and source records attributed to L C Rome.

43 records · Page 3Linked to original sources

PubMed1989

Histochemical and molecular determination of fiber types in chemically skinned single equine skeletal muscle fibers.

Until now, there has been no reliable method for histochemical determination of fiber types of single skinned muscle fibers. The major problem arises from the fact that most histochemical techniques use cross-sections of a large group of fibers and compare a given fiber with those surrounding it. This is not possible with a single skinned fiber which has been separated from a bundle to be used for mechanical analysis. A further problem is that the skinning procedure itself may alter the staining pattern. We have developed a procedure by which multiple cross-sections of single skinned fibers can be exposed to various histochemical reactions and the staining patterns compared on the same slide to those of frozen muscle and skinned bundles. By this procedure, three fiber types were distinguished by both Ca2+-ATPase and SDH reactions. The fiber typings determined from both enzyme systems correlated well with each other. Although we were able to differentiate only between slow and fast fibers by SDS-PAGE, these results corroborated the histochemical classification. This procedure will clearly be useful in skinned single muscle fiber mechanics experiments performed to determine functional differences among fiber types.

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PubMed1988

Why animals have different muscle fibre types.

Animals have different muscle fibre types: slow fibres with a low maximum velocity of shortening (Vmax) and fast fibres with a high Vmax. An advantage conferred by the use of different fibre types during locomotion has been proposed solely on the basis of their in vitro properties. Isolated muscle experiments show that force generation, mechanical power production and efficiency are all functions of V/Vmax, where V is the velocity of muscle shortening. But it is not known whether animals actually use the different fibres at shortening velocities that are optimal for mechanical power production and efficiency. Here we compare the V of muscle fibres during locomotion with their Vmax. This comparison shows that during slow locomotion, the slow fibres shorten at a velocity that gives peak mechanical power and efficiency and the fast fibres shorten at their optimal velocity when powering maximal movements. Our results also show that maximal movements are impossible without fast fibres because the slow ones cannot shorten rapidly enough.

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PubMed1986

The maximum speed of shortening in living and skinned frog muscle fibres.

This study was performed to determine whether Viso (the maximum speed of shortening extrapolated from force-velocity curves) equalled Vu (the unloaded speed of shortening determined by the slack test) in both living fibres from Rana temporaria and mechanically skinned fibres from Rana pipiens. In living fibres (R. temporaria) we obtained improved estimates of Viso by performing force clamps (isotonic) and length ramps (isovelocity) down to very low loads (0.005 isometric tension, P0). Force-velocity characteristics determined by force clamps and length ramps were the same. The hyperbolic Hill curves deviated from the force-velocity data at both high and low loads and underestimated Viso by varying degrees. A better estimate of Viso was obtained by linear extrapolation of data at loads from 0.005-0.02 P0 and the mean Viso at 7.5 degrees C was 4.08 muscle lengths/s +/- 0.11 (mean +/- S.E., n = 14). Improved estimates of Vu in living fibres were obtained by photographically calibrating the slack test. The mean Vu was 4.05 muscle lengths/s +/- 0.13 (mean +/- S.E., n = 14) and the intercept was 0.0156 fibre lengths (L0) +/- 0.0013 (mean +/- S.E., n = 14). The step-ramp photographic method, in which the motor speed is matched to Vu, was developed as an independent way to measure Vu in living fibres. Vu measured in this way agreed well with Vu measured by the slack test. In all living fibres, the improved estimates of Vu agreed well with the improved estimates of Viso. Vu/Viso = 0.99 +/- 0.01 (mean +/- S.E., n = 14). In mechanically skinned R. pipiens fibres, force clamps were performed down to loads of 0.01 mN. The force-velocity curve of the skinned fibres differed in shape from that of the living fibres. Although there was significant deviation from the Hill equation at low loads, the data at high loads were well fitted by the Hill curve. Viso determined by extrapolating the Hill equation to zero load was 5.87 muscle lengths/s +/- 0.38 (mean +/- S.E., n = 9) at 7.5 degrees C. In five fibres, the linear extrapolation of low loads (0.01-0.05 P0) showed that the Hill equation underestimated the true Viso by 6%. The slack test with mechanically skinned fibres was calibrated by taking a series of photographic exposures of the fibre at various times following each length step. Vu = 6.12 muscle lengths/s +/- 0.44 (mean +/- S.E., n = 10) and the intercept was 0.0585 L0 +/- 0.0069 (mean +/- S.E., n = 10).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1986

The influence of free calcium on the maximum speed of shortening in skinned frog muscle fibres.

The influence of [Ca2+] on the maximum velocity of shortening (Vmax) was examined in mechanically skinned Rana pipiens and Rana temporaria fibres using improved force clamps and the slack test techniques. All measurements were made at 7.5 degrees C. At low relative loads (P/P0 less than 0.1), maximally activated R. pipiens fibres shortened more rapidly than did submaximally activated fibres. At higher relative loads, however, little difference in the speed of shortening was observed. Vmax (determined by the slack test) of R. pipiens fibres increased as the level of activation increased. Over sarcomere lengths 1.8-2.1 microns it was 2.28 muscle lengths/s (m.l./s) (S.E. of mean +/- 0.25, n = 5) at 20-35% activation, 2.89 m.l./s (+/- 0.22, n = 7) at 40-60% activation, and 4.18 m.l./s (+/- 0.25, n = 6) at 100% activation. At longer sarcomere lengths (2.2-2.6 microns), higher Vmax values were observed at all levels of activation, but the influence of Ca2+ on Vmax persisted. Vmax was 3.54 m.l./s (+/- 0.41, n = 4) at 20-30% activation and 5.15 m.l./s (+/- 0.22, n = 5) at 100% activation. In R. temporaria fibres, Vmax (determined by force clamps over sarcomere lengths 1.8-2.1 micron) also increased as the level of activation increased, from 3.47 m.l./s (+/- 0.06, n = 6) at 13-29% activation to 5.62 m.l./s (+/- 0.17, n = 6) at 100% activation. Vmax was also determined (using the slack test) in mechanically and chemically skinned rabbit soleus fibres. Vmax at 15 degrees C (1.05 m.l./s, +/- 0.11, n = 5) at full activation decreased by more than 3-fold as the level of activation was reduced to 10%. We conclude that the level of activation influences the Vmax of skinned skeletal muscle fibres. This has now been demonstrated in three different preparations and by a variety of techniques. This effect is most pronounced at low relative loads, and might not be observed if there are experimental limitations which prevent making velocity measurements at low relative loads.

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PubMed1985

Stimulation rate, potentiators, and sarcomere length-tension relationship of muscle.

Isometric tetani of single muscle fibers of Rana temporaria were studied as a function of stimulation rate, sarcomere length (1.7-2.3 micron), twitch-to-tetanus ratio, and exposure to twitch potentiators (Zn2+ and NO3-) at 20 degrees C. As the stimulation rate was decreased below a maximal level, tension generation decreased. This depression in tension generation was more pronounced at shorter sarcomere lengths. Therefore the magnitude and shape of the sarcomere length curve was dependent on stimulation rate. Although the depression in tension generation was always accompanied by a noticeable ripple in the tension record in fibers with large twitch-to-tetanus ratios, it could be observed even during well-fused tetani in fibers with low twitch-to-tetanus ratios. In all fibers, however, high stimulation rates or exposure to potentiators resulted in maximum tension generation at each length, and the sarcomere length-tension curve followed that found by Gordon, Huxley, and Julian. This indicates that the fall in tension between sarcomere lengths of 2.0 and 1.7 micron is not due to length-dependent activation but is more likely to be the result of mechanical interference in the force-generating interaction between cross bridges and thin filament sites.

Animals↗

PubMed1984

Muscle fiber activity in carp as a function of swimming speed and muscle temperature.

These experiments were performed to determine how ectothermal animals maintain their locomotory ability during acute changes in muscle temperature, despite the large thermal dependence of the mechanical properties of their muscle. The electrical activity of the red (slow-twitch) and white (fast-twitch) muscle fibers of carp was monitored while the carp swam at various speeds at 10 and 20 degrees C. The patterns of recruitment of different fiber types were similar at both temperatures. At low speeds only the red muscle was active, whereas at high speeds the white muscle was active as well. The swimming velocity at which white muscle was initially recruited increased from 26 cm/s at 10 degrees C to 46 cm/s at 20 degrees C. These results suggest that the order of recruitment of motor units was the same at 10 and 20 degrees C but that the recruitment occurred over a narrower range of speeds at the low temperature. Hence, to generate the muscle power required to swim at a certain velocity, fish recruit more muscle fibers, which include faster fiber types when their muscle is cold than when their muscle is warm.

Animals↗

PubMed1983

Energetics of isometric contractions as a function of muscle temperature.

The energetic cost of generating isometric force in isolated frog muscle was examined at 10, 20, and 30 degrees C. Recovery O2 consumption (delta O2) and recovery lactate production (delta Lact) were measured under conditions in which O2 was not limiting metabolism. Both increased linearly with the force-time integral (integral of Fdt) generated by the muscle. The slopes of the regression equations for both delta O2 and delta Lact as a function of integral of Fdt increased with increasing temperature with a temperature coefficient (Q10) near 3. Total high-energy phosphate resynthesis from recovery metabolism was calculated by scaling the delta O2 regression equation and the delta Lact regression equation into equivalent ATP units and summing them. This total recovery metabolism was modeled as the sum of two components, a "cost of maintaining force" (slope of the equation) and a saturable "start up cost" (intercept of the equation). The cost of maintaining force increased with temperature with a Q10 near 3 over the whole temperature range, whereas the start up cost was nearly independent of temperature between 0 and 20 degrees C and fell to near zero at 30 degrees C. Delta O2 measurements from a series of tetani given in rapid succession showed that for contractions subsequent to the first, no start up cost was incurred and that the "cost of generating force" for these contractions was equal to the slope of the regression line for single tetani. The practical consequence of these facts is that, in series of tetani, the cost of generating force increases with a Q10 of 3.

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