The formation of synapses in reinnervated and cross-reinnervated striated muscle during development

1. A study has been made of the formation of synapses in reinnervated and cross-reinnervated developing striated muscles which normally receive either a focal or distributed innervation, using histological, ultrastructural and electrophysiological techniques.2. The focally innervated mammalian tibialis anterior muscle, denervated soon after birth, was reinnervated at both the original end-plates as well as on the new muscle added during the period of denervation; but not on the muscle present at the time of denervation. Nearly all the synapses which had formed, other than at the original end-plates, disappeared by 6 weeks post-natal.3. The avian anterior latissimus dorsi muscle (ALD), which receives a distributed innervation, was denervated during the first week post-hatched, and became reinnervated both at the original synaptic sites as well as on the new muscle added during the period of denervation; all these synapses were spaced approximately 200 mum apart along the length of individual muscle cells.4. The myofibres of the ALD muscle cross-reinnervated at hatching with the superior brachialis nerve, which contains fast motor axons that normally form a focal innervation, were each focally innervated by a single ;en plaque' terminal; these synapses had the same electrical properties as normal synapses formed by fast motor axons.5. Many of the myofibres of the avian posterior latissimus dorsi (PLD), which normally receive a focal innervation, received a distributed innervation from ;en grappe' terminals when cross-reinnervated with the ALD nerve at hatching.6. It is suggested that during development the nerve type determines the pattern of synapses over an effector; this is achieved by the nerve, after forming the initial synaptic contact, making the rest of the muscle cell membrane refractory to further synapse formation for some distance, this distance being determined by the nerve type.     

Changes in chemosensitivity of developing chick muscle fibres in relation to endplate formation

1. The sensitivity to acetylcholine (ACh) of the fast posterior latissimus dorsi (PLD) and slow anterior latissimus dorsi (ALD) during embryonic development was studied and compared. The sensitivities were expressed as a ratio of the maximal tetanic tension and tension developed in response to ACh. 2. Up to the 17th day of incubation both muscles are sensitive to ACh to a similar extent; at the 18th day the sensitivity of the PLD muscle decreases and continues to do so until hatching and thereafter. 3. Since the decrease in sensitivity of PLD muscles takes place a few days after innervation, it is suggested that this is caused by activity of the motor nerve. To test this curare (dTc) and hemicholinium (HC-3), drugs that interfere with neuromuscular transmission, were injected into the yolk sac of the embryos when nervemuscle connections are usually established. In the curare and HC-3 treated embryos the desensitization of the PLD muscles did not take place. 4. The distribution of endplates on PLD muscles from drug treated 20-21 day old embryos was compared to that of untreated controls. Whereas control PLD muscles have only one band of endplates, muscles from curarized embryos and HC-3 treated embryos have several bands of endplates, and many muscle fibres with multiple innervation were found. 5. It is suggested that nerve fibres which make connections with PLD muscle fibres bring about a decline in chemosensitivity by releasing more transmitter, and thereby prevent further nerve muscle connections from being made along the same muscle fibre.     

Fast and slow muscles of the chick after nerve cross-union

1. The multiply innervated anterior latissimus dorsi (ALD) and the focally innervated posterior latissimus dorsi (PLD) muscles of the chick were investigated 2-18 months after nerve cross-union.2. The fast PLD muscle re-innervated by the slow muscle nerve became supplied with ;en grappe' end-plates and responded to a single nerve volley with local potentials only. Control PLD muscles re-innervated by the original nerve, had ;en plaque' end-plates and responded to a single nerve volley by synchronous action potentials in the same way as normal muscles.3. In the slow ALD muscle re-innervated with the ;mixed' PLD nerve, the type of innervation and of electromyographic response remained practically unchanged, with the exception of transplanted ALD muscles supplied with PLD nerves where, in addition to local responses, propagated action potentials were registered electromyographically in response to single nerve volleys.4. ALD muscles of young chickens re-innervated both with an implanted purely fast muscle nerve and with the regenerated original nerve, had two types of innervation: ;en plaque' end-plates around the nerve implant, and multiple ;en grappe' end-plates in areas supplied with the ALD nerve. Accordingly, propagated action potentials were registered in response to single nerve volleys in regions near the implant, whereas local potentials were recorded in areas with original innervation.5. Contraction velocity was not substantially altered in PLD and ALD muscles after nerve cross-union.6. No changes were observed in the fine structure of muscle fibres in extrajunctional regions.     

Early morphological changes in the striated muscles in normal and dystrophic chickens

Histological and histochemical analyses were performed on the anterior latissimus dorsi muscle (ALD, red muscle) and the posterior latissimus dorsi muscle (PLD, white muscle) in normal (line 412) and dystrophic chickens (line 413) from 19 day embryos to 6 weeks of age. PLD, the white muscle, in dystrophic chickens showed higher percentages of red and intermediate fibres than those of normal chickens during the early development of muscles. Increases of the oxidative enzyme activities and the numbers of NADH--TR formazan granules in the white fibres of PLD were already found at 1 week of age in dystrophic chicken. Fibre types, oxidative enzyme activities and NADH--TR formazan granules showed no differences in ALD between normal and dystrophic chickens. These results suggest that increases of oxidative enzyme activities and formazan granule numbers and incomplete fibre type differentiation in PLD of dystrophic chickens are early pathological processes in such birds.     

The formation of synapses in mammalian sympathetic ganglia reinnervated with preganglionic or somatic nerves

1. A study has been made of the formation of synapses in the superior cervical ganglion of the guinea-pig, during reinnervation either with axons of the cervical sympathetic trunk, or with somatic axons of the nerve to the sternohyoid muscle.2. No significant changes in either the geometry or electrical parameters of sympathetic motorneurones were detected following denervation for periods of 3-6 weeks, or after reinnervation with either preganglionic or somatic axons.3. The post-ganglionic action potential reappeared about 4 weeks after preganglionic trunk section (eighteen of eighteen ganglia); its amplitude increased progressively and was almost normal by more than 10 weeks after nerve section. A very small response was detected from thirteen of eighteen ganglia after periods longer than 8 weeks after cross-reinnervation with somatic axons.4. Regenerated preganglionic or somatic nerve terminals were demonstrated around the ganglion cells using ZIO impregnation and electron-microscopy; the structure of these terminals was unchanged following regeneration into the ganglia, although many more synapses were formed by preganglionic terminals than by somatic terminals.5. The facilitation of evoked synaptic potentials which occurs during repetitive stimulation of preganglionic axons was retained following their regeneration, whereas most synapses formed on ganglion cells by regenerating somatic axons showed facilitation of transmitter release during trains of stimuli, rather than the normal depression.6. These observations suggest that the structure and electrical properties of adult mammalian autonomic motorneurones are not under neural control, but that these neurones do show some selectivity in the type of nerve which they will permit to form synapses on them.     

Development of neuromuscular junctions of fast and slow muscles in the chick embryo: a light and electron microscopic study

Summary The morphogenesis of neuromuscular junctions (NMJ) was studied by electron microscopy in fast posterior and slow anterior latissimus dorsi muscles (PLD and ALD) of chick embryos. In 8 day embryos, the NMJ is already established in both types. In PLD at this stage, individual axons completely ensheathed by Schwann cell processes form NMJs with myotubes, whereas in ALD axon terminals lie adjacent to (and not separated by Schwann cell processes from) naked axons which are components of a nerve bundle. At 11–15 days, the number of profiles of axon terminals at each endplate increases in both PLD and ALD. In PLD, individual axon terminals are ensheathed by Schwann cells and often branch. In ALD, several axon terminals become ensheathed as a group by processes of a Schwann cell, along with a small number of adjacent naked axons. The individual axon terminals were confirmed by analysis of serial sections to originate from different preterminal axons. Thus, the increase in number of axon terminals in PLD may be due to extensive terminal branching, whereas in ALD it may be due to the arrival of other nerve fibres. From 16 days, each axon terminal in an endplate of ALD becomes individually ensheathed by Schwann cell processes. However, the property of polyneuronal innervation in each endplate is retained even in the adult muscle. The junctional sarcoplasm protrudes to separate individual axon terminals at certain developmental stages: the protuberances are thinner and more numerous in ALD than in PLD at 15–16 days.It is concluded that NMJ morphogenesis differs between PLD and ALD and that the differences reside mainly on the neuronal side.     

Structure, distribution and innervation of muscle spindles in avian fast and slow skeletal muscle

Muscle spindles in 2 synergistic avian skeletal muscles, the anterior (ALD) and posterior (PLD) latissimus dorsi, were studied by light and electron microscopy to determine whether morphological or quantitative differences existed between these sensory receptors. Differences were found in the density, distribution and location of muscle spindles in the 2 muscles. They also differed with respect to the morphology of their capsules and intracapsular components. The slow ALD possessed muscle spindles which were evenly distributed throughout the muscle, whereas in the fast PLD they were mainly concentrated around the single nerve entry point into the muscle. The muscle spindle index (number of spindles per gram wet muscle weight) in the ALD was more than double that of its fast-twitch PLD counterpart (130.5±2.0 vs 55.4±2.0 respectively, n=6). The number of intrafusal fibres per spindle ranged from 1 to 8 in the ALD and 2 to 9 in the PLD, and their diameters varied from 5.0 to 16.0 μm and 4.5 to 18.5 μm, respectively. Large diameter intrafusal fibres were more frequently encountered in spindles of the PLD. Unique to the ALD was the presence of monofibre muscle spindles (12.7% of total spindles observed in ALD) which contained a solitary intrafusal fibre. In muscle spindles of both the ALD and PLD, sensory nerve endings terminated in a spiral fashion on the intrafusal fibres in their equatorial regions. Motor innervation was restricted to either juxtaequatorial or polar regions of the intrafusal fibres. Outer capsule components were extensive in polar and juxtaequatorial regions of ALD spindles, whereas inner capsule cells of PLD spindles were more numerous in juxtaequatorial and equatorial regions. Overall, muscle spindles of the PLD exhibited greater complexity with respect to the number of intrafusal fibres per spindle, range of intrafusal fibre diameters and development of their inner capsules. It is postulated that the differences in muscle spindle density and structure observed in this study reflect the function of the muscles in which they reside.     

The influence of innervation on the differentiation of contractile speeds of developing chick muscles

1. The role of innervation of the differentiation of contractile speeds was studied in the slow anterior latissimus dorsi (ALD) and fast posterior latissimus dorsi (PLD) muscle of the chick. 2. These muscles become innervated during the 12th and 15th day of embryonic development. At this time both muscles contract and relax extremely slowly and their contractile speeds are very similar. By the 18th day their contraction and relaxation becomes more rapid. It is at this time that the contractile characteristics of both muscles also become very different from each other, ALD being about 3 times slower than PLD. Thus innervation percedes differentiation of contractile speeds by several days. 3. The influence of innervation on the contractile characteristics of developing slow and fast muscles was studied during muscle regeneration in adults. When a slow ALD muscle was minced and implanted in place of a fast PLD the newly regenerated ALD became innervated by a PLD nerve and resembled a fast PLD. Conversely, when PLD muscles were minced and replaced ALD the regenerated PLD was innervated by ALD nerve and became slow. 4. Histological examination revealed that the regenerated ALD became focally innervated, and the regenerated PLD multiply innervated. 5. Thus, the contractile speeds are not predetermined properties of the muscle fibre. Both contractile characteristic and the pattern of innervation of developing muscles are determined by the motor nerve.     

The formation of synapses in amphibian striated muscle during development.

1. A study has been made of the formation of synapses in developing reinnervated and cross-reinnervated amphibian twitch muscles which receive either a focal (iliofibularis) or a distributed (sartorius) innervation from 'en plaque' nerve terminals using histological, ultrastructural and electrophysiological techniques. 2. During the development of the tadpole through metamorphosis to the adult frog, the sartorius myofibres increased in length at about twice the rate of the iliofibularis myofibres, due to a fast rate of growth at their insertions on to the pelvic tendon. 3. The short iliofibularis and sartorius myofibres of young tadpoles (800 mum long) possessed only a single synapse and the iliofibularis myofibres did not receive any further innervation during development. However the sartorius myofibres received further transient innervation on the new muscle laid down during development at the fast growing pelvic insertion, until the distance between the original synapse formed on the myofibres and the synapse at the pelvic end of the muscle was about 12 mm. 4. During development synapses possessed either skewed, multimodal, or unimodal m.e.p.p. amplitude-frequency distributions; the intervals between m.e.p.p.s. were not distributed randomly according to a Poisson process, as m.e.p.p.s. of similar amplitudes tended to be separated by very short intervals; the unit-size e.p.p. had a similar amplitude-frequency distribution as the m.e.p.p.s. if these had a unimodal distribution. 5. Reinnervation or cross-reinnervation of the sartorius and the iliofibularis muscles in adults or at a late stage of development simply reconstituted the normal focal and distributed innervation patterns of the muscles, as found in the control muscles of the contralateral and unoperated legs. 6. These observations on synapse formation in amphibia are consistent with the hypothesis that during development the axon making the initial synaptic contact on the muscle cells induces a property over a length of muscle membrane adjacent to this site which makes it refractory to synapse formation; thus during reinnervation or cross-reinnervation of adult muscles this refractory property constrains synapse formation to these sites.     

The formation of synapses in reinnervated mammalian striated muscle

1. The hemidiaphragm of the adult rabbit has a single band of end-plates running around the middle of the muscle. A study has been made of the formation of synapses during spontaneous reinnervation of this muscle, using histological, ultrastructural and electrophysiological techniques.2. Following spontaneous reinnervation, silver-stained nerve terminals were found in association with cholinesterase-stained end-plates only in the region of the muscle corresponding to the original innervation band.3. The regenerated nerve terminals were observed with the electronmicroscope in positions overlying or adjacent to the old synaptic folds.4. Spontaneous miniature end-plate potentials and evoked synaptic potentials were recorded only in the middle of the muscle fibres after reinnervation.5. The growth of the regenerating axons was not oriented towards the end-plate zone but followed muscle fibres and blood vessels in random directions.6. It is concluded that, in adult mammalian striated muscle, the old end-plate region is preferentially reinnervated as a consequence of some special property of the muscle fibre at this site.     

Characterization of Ca2+- and Sr2+-activated tension in functionally skinned chicken fibers of normal and dystrophic skeletal and normal cardiac muscle

The Ca²⁺ and Sr²⁺ activation of tension in functionally skinned chicken fibers of normal and dystrophic skeletal and normal cardiac muscle were studied. The muscles studied can be separated into two groups based upon their Ca²⁺ and Sr²⁺ sensitivities: those which are significantly more sensitive to Ca²⁺ than to Sr²⁺, pectoralis and posterior latissimus dorsi (PLD), and those which show no Ca²⁺/Sr²⁺ sensitivity difference, cardiac and anterior latissimus dorsi (ALD). This suggests that there is more than one type of Ca²⁺ site involved in Ca²⁺ control of muscle contraction in different muscle types and suggests that ALD and cardiac muscle may be controlled by a different type of binding site than PLD and pectoralis muscle. Dystrophic ALD and PLD muscles showed little change in their Ca²⁺ and Sr²⁺ sensitivities from those of normal muscles in contrast to the pectoralis which showed a decrease in both Ca²⁺ and Sr²⁺ sensitivity (approaching that of PLD) with the onset of dystrophy. Similarly, upon SDS polyacrylamide gel electrophoresis, dystrophic ALD and PLD muscles showed no difference in contractile proteins from those of normal muscles, in contrast to pectoralis muscle where the appearance of a 36,000 dalton protein band correlated with the onset of dystrophy and the changes in the Ca²⁺/Sr²⁺ activation properties of this muscle. The contractile protein band pattern of normal and dystrophic PLD and dystrophic pectoralis muscle were similar including the presence of the 36,000 dalton protein.     

Morphogenesis of rat muscle spindles after nerve lesion during early postnatal development

Summary The influence of innervation on muscle spindle morphogenesis has been investigated in rat hind-limb muscles by sectioning the sciatic nerve, with suture of the stumps, at various postnatal stages. After nerve section at 4 or 7 days of age a proportion of spindles survived during the denervation phase and developed, during the subsequent reinnervation phase, into atypical structures. The reinnervated spindles were recognized by the presence of a limiting capsule but lacked the characteristic distinction of equatorial and polar regions. The intrafusal fibres were fewer than normal and were indistinguishable in size and fine structure from extrafusal fibres; they had a single motor endplate and lacked sensory nerve terminals. In reinnervated muscles of animals operated at 13 and 22 days of age there was a progressive tendency towards a restoration of normal spindle structure and innervation. These findings indicate that muscle spindle morphogenesis is profoundly altered by nerve lesion at early developmental stages, apparently as a result of inadequate sensory reinnervation. This study also shows that the differentiation of intrafusal fibres is dictated by their specific pattern of innervation and is not intrinsically predetermined.     

Matching of facilitation at the neuromuscular junction of the lobster: a possible case for influence of muscle on nerve

1. The facilitation of neuromuscular transmission, which occurs during repetitive activation, was examined in the proximal accessory flexor muscle in walking legs of the lobster using electrophysiological techniques.2. Post-synaptic potentials (p.s.p.s) in different muscle fibres facilitated to markedly different degrees. P.s.p.s in some fibres did not facilitate at all, while in others they increased in size by 20-30 times during stimulation at 20 Hz even though all the excitatory neuromuscular synapses are made by a single axon.3. Stimulation of widely separated groups of synapses on any single muscle fibre evoked p.s.p.s with closely matched facilitation properties. Extracellular p.s.p.s recorded from single synaptic spots showed the same characteristics of facilitation as those of intracellular p.s.p.s in the same muscle fibre, suggesting that individual synaptic contacts on any single fibre are similar to each other.4. Facilitation can be accounted for by an increase in the number of quanta released from the nerve terminals. There is no evidence for an increase in post-synaptic membrane sensitivity.5. Low Ca solutions reduce transmitter release with comparatively little change in facilitation, while Cs solutions increase the size of p.s.p.s without increasing the amplitude of spontaneous miniature potentials. Thus, at poorly facilitating synapses it is unlikely that the absence of facilitation is caused by the saturation of some post-synaptic process.6. It is concluded that the excitatory presynaptic nerve terminals on a single muscle fibre have matching facilitation characteristics. Some interaction between individual muscle fibres and their associated nerve endings may be required to establish or maintain this matching.     

The influence of secretin on the secretion of pepsin in response to acid stimulants in the anaesthetized cat.

1. The problem of selectivity during reinnervation of skeletal muscle fibres was investigated in the rat using the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscles and their nerves. 2. After an operation on these nerves permitting them to compete for reinnervation of one or the other muscle (hereafter called Y-union), virtually the total isometric tetanic tension of EDL muscle could be elicited by stimulating the EDL nerve, while stimulating the soleus nerve yielded little or no tension. In the case of the soleus muscle, stimulation of either nerve elicited about half of the total isometric tetanic tension. 3. During the course of reinnervation of these muscles in non-competitive situations, the time course of increase in the ratio of tension elicited by nerve stimulation to that by direct stimulation was slower in the case of soleus nerve reinnervating EDL muscle, compared with cross-reinnervation in the reverse direction or reinnervation of each muscle by its own nerve. 4. Crushing the common peroneal nerve 12 days after a Y-union in an attempt to retard the EDL nerve did not favour reinnervation of the EDL by soleus nerve, but crushing the nerve again or just once at 1 month after the original operation produced substantial partial reinnervation of the EDL by the soleus nerve. 5. It is concluded that soleus nerve fibres form functioning neuromuscular synapses on EDL muscle fibres only with difficulty. The pattern of reinnervation reveals characteristic differences between fast-twitch and slow-twitch muscles on the one hand and between their respective nerves on the other.     

The onset and progress of transformation of avian slow into fast muscles under neural influence

The slow anterior latissimus dorsi (ALD) muscles of newly hatched chickens were transposed and cross0innervated by the mixed, predominantly fast superior brachialis nerve, and investigated 2 to 15 months after the operation. Two months after the operation, myosin ATPase activity of the cross-innervated ALD muscles was still as low as in the control ALD, although the ultrastructure and the histochemical ATPase activity already showed a mixed fibre-type pattern with a predominance of fast -type fibres around the site of nerve implantation. The change of myosin properties of thw whole cross-innervated ALD did not occur until the third month after the operation. At that time, the myosin ATPase activity increased about 2.5 times and light chains of myosin of the fast type appeared in the electrophoretic pattern. The myosin ATPase activity attained 62% of the activity found in the control fast posterior latissimus dorsi muscles at three months; subsequently it remained at about this level reaching 68% 18 months after the operation. The results indicate that approximately two thirds of the cross-innervated ALD muscle fibres became changed towards the fast type under neural influence, whereas about one third remained slow, being re-innervated by the slow-type motor fibres of the implanted nerve.     

The role of muscle activity in the differentiation of neuromuscular junctions in slow and fast chick muscles

Summary The differentiation of neuromuscular junctions of multiply innervated, slow, anterior latissimus dorsi (ALD) and focally innervated, fast, posterior latissimus dorsi (PLD) muscles was studied in normal and curarized chick embryos. At 16 days of incubation, fibres of both muscles are contacted by several axon profiles, the number of which falls with age. In 18-day-old embryos individual endplates in ALD are usually contacted by three axon profiles, whereas in PLD, endplates are contacted only by a single large terminal profile. At this time, there is already a significant accumulation of cell organelles in the postsynaptic area.Treatment of embryos with curare during the 7th and 12th day of incubation delays the differentiation of the neuromuscular junction in both muscles. The paralysis dramatically affects the decrease of the number of axon profiles at individual endplates in both muscles. At 16 days the number of axon profiles was greater in embryos treated with curare than in the untreated controls. At 18 days when the number of axon profiles normally decreases, the endplates of both types of curarized muscles have an even greater number of axon profiles than at 16 days. Endplates in curarized PLD had up to 13 and in curarized ALD up to 12 axon profiles. The effects of curare gradually wore off and when the movements of the embryos again became more vigorous, the normal differentiation of neuromuscular junctions continued. At 21 days of incubation many embryos recover from curare and show endplates of normal appearance in both muscles. These results suggest that activity of the muscle is essential for the maturation of the neuromuscular junctions.     

Differentiation of membrane systems during development of slow and fast skeletal muscle fibres in chicken

Summary The disposition of transverse (T) tubules, sarcoplasmic reticulum (SR) and T-SR junctions (triads) and the width of Z lines are matched to contractile properties in adult muscle fibres. We have studied the development of the membrane systems in the slow anterior (ALD) and the fast posterior (PLD) latissimus dorsi of the chicken in ovo (E14–E21) and after hatching (D1–D30). T tubules, SR, triads and Z lines were visualized using DiIC16[3] labelling for confocal microscopy and either Ca-osmium-ferrocyanide or standard procedures for electron microscopy. Anterior latissimus dorsi and PLD have similar, slow twitches in early development (E14–E16), but PLD suddenly becomes faster starting at E17–E18. We find that in coincidence with the differentiation of faster contraction properties (starting at E18–E19) density of triads is significantly higher and width of Z lines is narrower in PLD. The SR also begins to acquire fibre-type specific characteristics at this time. Early development of T tubules, on the other hand, is quite similar in the two muscles. Peripherally-located, longitudinally-oriented T tubules, and the first T networks crossing the fibre center appear earlier in ALD (E14–E15 and E16) than in PLD (E14–E16 and E17), but have similar dispositions. The final fibre-type specific distribution of T tubules is achieved after hatching. Some T tubules-rich fibres in the ALD, presumably future fast fibres, develop extensive T tubule networks at early stages. Location of triads at the Z line in pectoralis occurs in three steps: an initial location of longitudinally oriented triads at the A-I junction; a subsequent move to the Z lines and finally a rotation to a transverse orientation.     

Suppression of foreign innervation in axolotl muscle may not be dependent on juxtaposition of native and foreign nerve terminals.

The posterior supracoracoideus nerve of the axolotl, Ambystoma mexicanum, was induced to make synapses outside its normal muscle territory. Muscle fibres with inputs from both native and foreign nerves were studied during the period of suppression of foreign transmission and in only 8% of fibres were foreign and native terminals found within 120 micrometer of each other. A combined cholinesterase/silver staining technique revealed non-innervated endplates in foreign-innervated fibres just prior to the return of the native nerve. These results suggest a mechanism enabling suppression of foreign synapses at some distance from native synapses probably beginning with the reinnervation of empty sites by the native nerve.     

Freeze-fractured sarcoplasmic reticulum in adult and embryonic fast and slow muscles

Summary There is evidence to suggest that 8 nm calcium transport particles in the sarcoplasmic reticulum are involved in the regulation of twitch properties in adult muscles. We have studied ultrastructural characteristics of the sarcoplasmic reticulum in relation to previously defined physiological changes that take place in the normal course of development. The fast twitch posterior latissimus dorsi (PLD) and the slow tonic anterior latissimus dorsi (ALD) of the chicken were compared using the procedure of freeze-fracture. In the adult PLD, the sarcoplasmic reticulum was composed of longitudinal tubules, which gave rise to fenestrated cisternae at the centre of the H band and to terminal cisternae that form triads regularly at each A-I junction. In most of the fibres (85%), 8 nm intramembrane particles were closely packed in the concave fracture face (P-face). In the ALD, a tubular network with an open circular pattern extended the entire length of the A band and usually throughout the I band as well. Dyads or triads, which were infrequent, were often oriented obliquely. The density of intramembrane particles was low in the majority of the fibres, but there was a significant minority population (30%) in which particle density was relatively high. At 10 daysin ovo, when speed of contraction in both the ALD and PLD is slow, there was a circular configuration of sarcoplasmic reticulum components in both muscles, and particle density was low. Surprisingly, at 18 daysin ovo, when the rate of tension development and relaxation have reached nearly adult values in the fast PLD, this muscle, like the ALD, continued to exhibit a circular arrangement of sarcoplasmic reticulum tubules. The density of P-face particles, although greater than at 10 days, was still low relative to the adult PLD. Estimated values for the 18-day PLD were similar to those calculated for the adult slow muscle. Our observations, along with those of other investigators, suggest that abundant intramembrane particles may be related to the fast twitch properties of the adult PLD. However, they indicate that neither the pattern of membranes typical of the adult fast muscle nor the high content of calcium transport particles is required for the differentiation of fast twitch characteristics.     

Innervation of muscle receptors in the cross-reinnervated soleus muscle of the cat

It has recently been reported (Gregory et al., J. Physiol., 331:367-383, 1982) that cutting a muscle nerve and letting it grow back into the muscle or cross-uniting the muscle with a foreign nerve results in major disruption of the normal response patterns of muscle spindles and tendon organs. Here we report observations on the structure of muscle receptors in cross-reinnervated and self-reinnervated soleus muscles in an attempt to detect abnormalities that might account for their disturbed function. Eight soleus muscles were reinnervated with the extensor digitorum longus nerve for periods up to 449 days and two were self-reinnervated. Following the physiological investigation, the muscle was fixed and stained according to the method of Barker and Ip (J. Physiol., 69:73P-74P, 1963). Spindles and tendon organs were teased from the muscle and photographed. In one cross-reinnervated muscle an attempt was made to isolate all receptors. About two-thirds of the normal number of spindles and tendon organs were found. Three categories of receptor were identified: normal, abnormal, and those having no visible nerve endings. There appeared to be little difference in degree of abnormality of receptors in self- and cross-reinnervated muscles. Of the 180 spindles, 3% were normal, 43% had no visible endings, and 54% had abnormal endings. Of 80 tendon organs, 38% were normally innervated, 33% were without visible innervation, and 29% had abnormal endings. We conclude that following long-term cross-reinnervation and self-reinnervation of soleus there is extensive disruption of the normal innervation pattern of both spindles and tendon organs which could account for their functional abnormalities.