Individual differences in multiple-bag spindles of cat superficial lumbrical muscles.

A total of 791 spindle poles was analysed with regard to intrafusal fibre composition in the first and second superficial lumbrical muscles from the right and left hindfeet of 9 male and 5 female adult cats. Bag and chain muscle fibres were identified by their myofibrillar ATPase staining profile in the B region, after either acid or alkaline preincubation. A high proportion of the spindle pole population (43.2%) was observed to contain three or more (up to 5) bag fibres; those poles were classified as multiple-bag spindle poles. In the 334 muscle spindles in which both poles were studied, 42 bag fibres (12.6%) were found to be of the 'mixed' type, that is a fibre in which the two poles differ in their ATPase staining profile (either bag1/bag2 or bag/chain). The variability of the intrafusal fibre content observed in spindles of these muscles has been studied in relation to individual characteristics such as sex, weight and side of the animal. In general, multiple-bag spindles are more frequent in male than in female cats and in right as compared to left side muscles. Nearly all 'mixed' bag intrafusal fibres (38 out of 42) were observed in spindles containing 3 or more bag fibres. In 3-bag spindles the proportion of 'mixed' bag spindles is approximately the same in male and female cats. The ratio of 'dynamic' (mean polar bag1 content) to 'static' (mean polar bag2 plus chain fibre content) intrafusal effectors per muscle tends to increase in spindles of right side muscles and to decrease in the heaviest animals. The quantitative and qualitative differences in fibre content of spindles observed in first lumbrical muscles of different animals suggest that the spindle fibre composition, especially that of the 'dynamic' bag1 fibre, may be related to individual predetermined and/or acquired factors.

High incidence of long-chain fibers in spindles of cat superficial lumbrical muscles.

Forty-two complete spindle poles of cat superficial lumbrical muscles were analyzed with particular regard to the length and the diameter of intrafusal fiber types. Poles were reconstructed from serial transverse sections of fresh-frozen muscles. The staining module, which was repeated throughout the whole muscle, comprised sections treated for glycogen detection and sections treated for detecting myofibrillar ATPase activity after preincubation at three different pH's (see METHODS). The identification of intrafusal fiber types was essentially based on the ATPase activity of the B region of the intrafusal fibers. Long-chain fibers, i.e., chain fibers that have at least one pole that extends by more than one millimeter beyond the end of the spindle capsule (6), were very commonly observed. Of 42 spindle poles analyzed, 30 (71%) contained at least one long-chain fiber (one in 17 spindle poles, 2 in 11 poles, and 3 in 2 poles). Of 246 poles of chain fibers, 45 (18%) were "long". In four spindles, in which both poles could be completely examined, 10 long-chain fibers were observed. In eight of these, only one pole was long; the opposite pole ended either intracapsularly or at a short distance outside the capsule. Since long-chain fiber poles, presently considered to be among the effectors of static skeletofusimotor (beta) axons, are present in a large proportion of muscle spindles of lumbrical muscles, it would be of particular interest to reevaluate the beta-supply of these muscles by physiological methods.

Glycogen depletion elicited in tenuissimus intrafusal muscle fibres by stimulation of static gamma-axons in the cat.

In this study the experimental conditions used to elicit glycogen depletion in tenuissimus intrafusal muscle fibres were different from those used by Barker, Emonet-Dénand, Harker, Jami & Laporte (1976): the tenuissimus was left in situ; several (4-6) static gamma-axons were stimulated together; the blood flow through the muscle was not reduced during the periods of gamma stimulation except in two experiments; very much longer periods (up to 9 h) of intermittent stimulation by bursts at 50-500/s were used. Bag1 and bag2 fibres were identified by their different ATPase activities in the B region. In two experiments with normal circulation, test responses of several primary endings to short periods of stimulation at 50-100/s were still very strong after stimulation of several static gamma-axons for 5 and 9 h, respectively. Glycogen depletion was observed in a large number of chain and bag2 poles but in only one of nineteen bag1 poles examined. In two other experiments with normal circulation, there was a very pronounced reduction of the test responses after stimulation of several static gamma-axons for 7 and 9 h, respectively. Out of twenty-four bag1 poles examined, nineteen exhibited zones of depletion. In an experiment in which stimulation was conducted as in Barker et al. (1976), i.e. with reduction of muscle blood flow during 1 min periods of stimulation at 50-100/s, the primary endings still gave a strong response after fifteen periods of stimulation in contrast with the marked 'fatigue' that was constantly observed in the former study. No depleted intrafusal fibres were found in the spindles of this muscle. In a last experiment, after an initial pattern of stimulation similar to that described above, the new pattern of stimulation, but with periodical reduction of blood flow, was applied, leading to a 'fatigue' of the test responses in 2 h. In the spindles of this muscle three out of ten bag1 poles were depleted. The variability of glycogen depletion in bag1 fibres appears to be linked to the degree of spindle 'fatigue' which may develop after static gamma stimulation. It seems that in 'fatigued' spindles some factor or factors liberated by the contraction of neighbouring fibres may deplete glycogen in bag1 fibres by a non-neural mechanism. When, in spite of a prolonged stimulation of static gamma-axons, no fatigue of the test responses develops, zones of depletion in bag1 fibres--possibly of neural origin--are very rare, although a large proportion of bag2 and chain fibres are depleted.

A study of mammalian intrafusal muscle fibres using a combined histochemical and ultrastructural technique.

A direct correlation of the histochemical and ultrastructural properties of intrafusal muscle fibres has been achieved by cutting frozen serial sections for histochemical applications (15 micron thick sections) and for electron microscopy (60 micron thick sections) in a repeating sequence. Three types of intrafusal fibre were recognized, including two types of nuclear-bag fibre, designated bag1 and bag2. In addition to histochemical and ultrastructural differences, the three types of fibre differed in length and diameter. Regional variations of histochemical and ultrastructural properties were found. The results are compared with previous attempts to correlate histochemical and ultrastructural properties of intrafusal muscle fibres based on indirect methods.

Fast-conducting skeletofusimotor axons supplying intrafusal chain fibers in the cat peroneus tertius muscle.

1. In six experiments on cat peroneus tertius muscle, from 12 to 23 motor axons with conduction velocities above 85 m/s were repetitively stimulated so as to produce glycogen depletion in the muscle fibers they innervated. 2. The whole muscle was then quick-frozen, serially cut, stained to demonstrate glycogen, and examined for intrafusal glycogen depletion. 3. Zones of glycogen depletion were found in 27 of the 99 examined spindles: they were almost invariably located on chain fibers and specifically on the longest of the chain fibers in affected spindles. 4. Since it was shown that there are no purely fusimotor fast axons in the motor supply to peroneus tertius, it is concluded that skeletofusimotor axons are present among the fastest motor axons to this muscle.

Types of intra- and extrafusal muscle fibre innervated by dynamic skeleto-fusimotor axons in cat peroneus brevis and tenuissimus muscles, as determined by the glycogen-depletion method.

1. The types of intra- and extrafusal muscle fibre innervated by dynamic skeleto-fusimotor (beta) axons were determined by using a modification of the glycogen-depletion method of Edström & Kugelberg (1968) combined with histochemical tests for various enzyme reactions. A single beta axon was prepared in each of the experiments, which were carried out on six peroneus brevis and two tenuissimus muscles. 2. The intrafusal distribution of dynamic beta axons is almost exclusively restricted to bag1 fibres. The bags fibre was depleted in each of twenty-four beta-innervated spindle poles; the only fibres of a different type depleted intrafusally were a bag2 fibre in one pole and a long chain in another. 3. Depletion in the bag1 fibres was usually restricted to one zone in one pole, generally in a mid-polar location. 4. The extrafusal muscle fibres depleted by dynamic beta axons belong to the slow oxidative type as defined by Ariano, Armstrong & Edgerton (1973). The number of such fibres in each motor unit could not be accurately determined, but is almost certainly small. 5. The slow oxidative muscle fibres innervated by dynamic beta axons were not depleted over their entire length. Since there is no reason to assume that they are not twitch fibres, it would seem that the localized depletions result from the conditions required to obtain glycogen depletion, i.e. long periods of motor stimulation applied during the occlusion of the muscle's blood supply. Under similar experimental conditions depletion of glycogen was also restricted to portions of fibres in fast oxidative-glycolytic motor units, but extended over most of the length of the fibres in fast glycolytic units.

Distribution of fusimotor axons to intrafusal muscle fibres in cat tenuissimus spindles as determined by the glycogen-depletion method.

1. The distribution of fusimotor axons to bag1, bag2 and chain muscle fibres in cat tenuissimus spindles has been studied using a modification of the glycogen-depletion technique of Edstrrom & Kugelberg (1968). Single fusimotor axons were stimulated intermittently at 40-100/sec for long periods (30-90 sec) during blood occlusion. Portions of muscle containing the activated spindles were quick-frozen, fixed in absolute ethanol during freeze-substitution, and then embedded in paraffin wax. Serial transverse sections were stained for glycogen using the periodic acid-Schiff method, and examined for depletion. 2. Dynamic gamma axons (i.e. those that increase the dynamic index of primary-ending responses to ramp stretches of large amplitude) depleted bag1 fibres almost exclusively. 3. Static gamma axons (i.e. those that reduce or abolish the dynamic index) depleted both bag and chain fibres. Bag1 and bag2 fibres were depleted about equally. 4. A single static gamma axon may activate both bag and chain fibres in one spindle (the most common pattern), chain fibres only in another, and bag fibres only in a third spindle. 5. Static gamma axons with conduction velocities less than 25 m/sec also had a non-selective distribution, but no depletion was observed in bag2 fibres. 6. The zones of depletion produced by dynamic gamma axons were distributed more or less equally in the intra- and extracapsular parts of spindle poles, whereas those produced by static gamma axons were mainly intracapsular. 7. The results are compared with the glycogen-depletion studies of Brown & Butler (1973, 1975) and our own study of the distribution of static gamma axons to spindles in which all other motor axons had degenerated (Barker, Emonet-Dénand, Laporte, Proske & Stacey, 1973). The implications of the finding that both static gamma and dynamic gamma axons activate bag1 fibres are discussed.