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The interaction between foreign and original motor nerves innervating the soleus muscle of rats.

1. The fibular nerve was transplanted on to the soleus muscle of the rats. Interruption of the original soleus nerve then permitted cross-innervation, and subsequently, over a period of weeks, re-innervation by the original nerve. 2. Individual muscle fibres were often innervated by both the original and the foreign nerve. The original and foreign end-plates were located in separate regions of the muscle. There were no indications that the original nerve could displace or repress the foreign innervation. 3. The extent of re-innervation by the original nerve depended upon the method of denervation. A single crush of the nerve was followed by virtually complete re-innervation, even of muscle fibres already innervated by the foreign nerve. When re-innervation was delayed by resection of a segment of the nerve only muscle fibres without foreign nerve innervation were re-innervated. Denervation by a simple nerve cut gave an intermediate result. 4. Re-innervation by the original nerve can take place without measurable extrajunctional sensitivity to ACh. 5. The original end-plate region could retain high and localized sensitivity to ACh for several months despite degeneration of its motor nerve terminal and activity of the muscle fibre. 6. Established foreign end-plates were re-innervated by the foreign nerve on muscle fibres with intact original innervation. 7. The factors controlling synapse formation in skeletal muscles are discussed.

Acetylcholine↗

Polyneuronal innervation of skeletal muscle in new-born rats and its elimination during maturation.

1. The events taking place during the elimination of polyneuronal innervation in the soleus muscle of new-born rats have been studied using a combination of electrophysiological and anatomical techniques. 2. Each immature muscle fibre is supplied by two or more motor axons which converge on to a single end-plate. There was no sign of electrical coupling between muscle fibres receiving multiple synaptic inputs. By the end of the second week after birth virtually all muscle fibres are innervated by only a single motor axon. 3. The average tension produced by individual motor units, measured in terms of the percentage of the total muscle twitch tension, declined dramatically during the first 2 weeks after birth. During this period there was no significant change in the number of motor neurones innervating the soleus muscle. Thus, the disappearance of polyneuronal innervation reflects a decrease in the number of peripheral synapses made by each motor neurone. 4. The decline in motor unit size was delayed, but not ultimately prevented, by the early surgical removal of all but a few motor axons to the soleus muscle. This procedure also caused a delay in the removal of polyneuronal innervation involving the remaining motor units. 5. Following a crush of the soleus nerve in neonatal animals, regenerating axons usually returned to the original end-plates. Polyneuronal innervation was extensive at early stages of re-innervation and it disappeared during the second week after birth just as in normal muscles. 6. Cross-innervation of neonatal muscles by an implanted foreign nerve caused a rapid disappearance of cholinesterase at denervated original end-plates and in most fibres prevented re-innervation by the original nerve. In the small proportion of fibres that did become innervated through both the foreign and original nerves the end-plates were more than 1 mm apart, and both foreign and original nerve end-plates could persist indefinitely. 7. Many cross-innervated fibres received multiple inputs through the foreign nerve. Some foreign end-plates were separated by distances ranging up to 1 mm. Polyneuronal innervation through the foreign nerve was completely eliminated during maturation but over a slightly longer period than in normal muscles. Apparently the elimination process can act over a distance up to but not much more than 1 mm. 8. These observations suggest that there are several factors influencing the elimination of redundant inputs in immature muscles. Individual motor neurones appear to have an inherent tendency to withdraw the majority of their original complement of peripheral terminals. The determination of which particular synapses are to survive, however, seems to be made in the periphery by a selection among all the synapses that innervate a limited region of each muscle fibre. There may be a competitive interaction among synapses in which those belonging to smaller motor units are less likely to be eliminated, thereby leading to a relatively uniform size of the motor units in the soleus.

Animals↗

Afferent innervation of the utricular macula in pigeons.

Biotinylated dextran amine (BDA) was used to retrogradely label afferents innervating the utricular macula in adult pigeons. The pigeon utriclar macula consists of a large rectangular-shaped neuroepithelium with a dorsally curved anterior edge and an extended medioposterior tail. The macula could be demarcated into several regions based on cytoarchitectural differences. The striola occupied 30% of the macula and contained a large density of type I hair cells with fewer type II hair cells. Medial and lateral extrastriola zones were located outside the striola and contained only type II hair cells. A six- to eight-cell-wide band of type II hair cells existed near the center of the striola. The reversal line marked by the morphological polarization of hair cells coursed throughout the epithelium, near the peripheral margin, and through the center of the type II band. Calyx afferents innervated type I hair cells with calyceal terminals that contained between 2 and 15 receptor cells. Calyx afferents were located only in the striola region, exclusive of the type II band, had small total fiber innervation areas and low innervation densities. Dimorph afferents innervated both type I and type II hair cells with calyceal and bouton terminals and were primarily located in the striola region. Dimorph afferents had smaller calyceal terminals with few type I hair cells, extended fiber branches with bouton terminals and larger innervation areas. Bouton afferents innervated only type II hair cells in the extrastriola and type II band regions. Bouton afferents innervating the type II band had smaller terminal fields with fewer bouton terminals and smaller innervation areas than fibers located in the extrastriolar zones. Bouton afferents had the most bouton terminals on the longest fibers, the largest innervation areas with the highest innervation densities of all afferents. Among all afferents, smaller terminal innervation fields were observed in the striola and large fields were located in the extrastriola. The cellular organization and innervation patterns of the utricular maculae in birds appear to represent an organ in adaptive evolution, different from that observed for amphibians or mammals.

Animals↗

Innervation pattern of a pool of nine excitatory motor neurons in the flexor tibiae muscle of a locust hind leg

The flexor tibiae muscle of a locust hind leg consists of 10-11 pairs of fibre bundles in the main body of the muscle and a distal pair of bundles that form the accessory flexor muscle, all of which insert onto a common tendon. It is much smaller than the antagonistic extensor tibiae muscle and yet it is innervated by nine excitatory motor neurons, compared with only two for the extensor. To determine the pattern of innervation within the muscle by individual motor neurons, branches of the nerve (N5B2) that supplies the different muscle bundles were backfilled to reveal somata in the metathoracic ganglion. This showed that different muscle bundles are innervated by different numbers of excitatory motor neurons. Physiological mapping of the innervation was then carried out by intracellular recordings from the somata of flexor motor neurons in the metathoracic ganglion using microelectrodes. Spikes were evoked in these neurons by the injection of current, and matching junctional potentials were sought in fibres throughout the muscle using a second intracellular electrode. Each motor neuron innervates only a restricted array of muscle fibres and, although some innervate a larger array than others, none innervates fibres throughout the muscle. Some motor neurons innervate only proximal fibres and others only more distal fibres, so that the most proximal and most distal bundles of muscle fibres are innervated by non-overlapping sets of motor neurons. More motor neurons innervate proximal bundles than distal ones, and there are some asymmetries in the number of motor neurons innervating corresponding bundles on either side of the tendon. Individual motor neurons cause slow, fast or intermediate movements of the tibia, but their patterns of innervation overlap in the different muscle bundles. Furthermore, individual muscle fibres may also be innervated by motor neurons with different properties.

Journal Article↗

Acetylcholinesterase-positive and paraformaldehyde-induced-fluorescence-positive innervation in the upper eyelid of the sheep (Ovis aries).

This is the first study which describes the innervation of some eyelid structures, such as the glands of Moll and the glands of Zeiss. It is also the first to investigate the innervation pattern of the eyelid as a whole. We have studied the acetylcholinesterase-positive and paraformaldehyde-induced-fluorescence-positive (FIF+) innervation pattern of the different structures that constitute the upper eyelid of the sheep. There is widespread acetylcholinesterase-positive innervation in the epithelium, but not such an abundant FIF+ innervation. Both types of innervation are represented in the connective tissue by trunks or fibers that are distributed towards the different structures immersed within them. In the glands of Zeiss, cholinesterase-positive innervation is much more widespread than FIF innervation. On the contrary, the glands of Moll present denser FIF+ innervation than acetylcholinesterase-positive innervation. The Meibomian glands and the lachrymal glands show a rich acetylcholinesterase-positive and FIF+ innervation. Eyelid muscle innervation is mainly acetylcholinesterase-positive. In the conjunctive membrane there is no acetylcholinesterase-positive innervation, and only scarce FIF+ fibers can be demonstrated.

Acetylcholinesterase↗

[Innervation pattern to the extensor digitorum brevis by deep peroneal nerve and accessory deep peroneal nerve].

On the electrophysiological basis, extensor digitorum brevis(EDB) muscle is innervated electrophysiologically not only by deep peroneal nerve(DPN) but also by accessory deep peroneal nerve(ADPN), an anomalous branch of superficial peroneal nerve, with a prevalence of 17-28%. We investigated 23 patients who had both DPN and sufficient ADPN innervation to the EDB on the intramuscular distribution of DPN and ADPN innervation to the medial and lateral side of the EDB. Recording electrodes were placed on the medial and lateral edges of the EDB with a supramaximal stimulation to the anterior or lateral ankle, compound muscle action potential (CMAP) of DPN or ADPN innervation was recorded. In 19 patients (83%) the DPN innervation was larger than or equal to the ADPN innervation. Only in 4 patients (17%) the ADPN innervation obviously exceeded the DPN innervation. DPN enters to the EDB from the medial side, and ADPN from the lateral side of the EDB. In 16 patients(70%) the DPN innervation was relatively large and the ADPN innervation was relatively small at the medial side of the EDB, and vice versa at the lateral side of the EDB. These distributions were almost uniform in 5 patients(22%). This study clarified that a biased larger DPN innervation and smaller ADPN innervation to the medial side of the EDB, and vice versa to the lateral side of the EDB in the majority cases. In some cases diffuse innervation to the EDB was found.

Action Potentials↗

A comparative light microscopic analysis of the sensory innervation of the mystacial pad. II. The common fur between the vibrissae.

The innervation to the common fur between the vibrissae was examined in the hamster, mouse, rat, gerbil, rabbit, guinea pig, and cat. Samples were taken from central locations among the more caudal vibrissae in the mystacial pad and processed with Richardson's variant of the Bielschowsky silver technique or with Winkelmann's silver technique to selectively stain peripheral axons and terminals. Additional samples were taken among the rostral vibrissae in the rat. We found major unpredictable species-related variations in the distribution of receptor types, innervation density, and the quantity of innervation in the skin between neighboring vibrissae. The common fur is composed of numerous larger guard hairs and even more numerous smaller vellus hairs. The guard hairs usually are richly innervated with fully developed piloneural complexes composed primarily of a pallisade of lanceolate endings and a circumferential array of Ruffini and free nerve endings. The vellus hairs are usually innervated by individual or shared free nerve endings. The piloneural complexes in the cat, rat, and mouse are usually complete, whereas those in the other species were usually incomplete and lacked Ruffini endings. There is considerable interspecies variation in the relative quantity of innervation between homologous neighboring vibrissae. The quantity of innervation is related to a combination of receptor completeness, innervation density, and distance between vibrissae. The quantity of intervibrissal fur innervation is by far highest in the cat, relatively high in the rabbit, relatively low in the hamster and caudal mystacial pad of the rat, and lowest in the mouse, gerbil, guinea pig, and rostral mystacial pad of the rat. The differences in the innervation between the cat and the rabbit correlate well with published physiologic data on types of receptor units. Also, barrels are most prominent in species having relatively low quantities of intervibrissal innervation and are less prominent or absent in species having high quantities of intervibrissal innervation.

Animals↗

Innervation of extraocular muscles in the rabbit.

The innervation of extraocular muscles in the rabbit was studied by using the methods of horseradish peroxidase (HRP) histochemistry, gross dissection, and quantitative morphology. Subdivisions of the oculomotor complex that innervate the superior rectus, inferior rectus, medial rectus, and inferior oblique and levator palpebrae are described, and our results are in agreement with previous accounts of the projections of this nucleus. Our analysis of the innervation of the lateral rectus and retractor bulbi muscles, however, differs from previous descriptions. The axons of approximately 80% of neurons in the abducens nucleus are in the VIth nerve and innervate the lateral rectus muscle, and approximately 15-20% are internuclear neurons both surrounding and intermingling with the motor neurons of the abducens nucleus. The interneurons project to the medial rectus subdivision of the contralateral oculomotor complex via the medial longitudinal fasciculus (MLF). Neurons in both the abducens and the accessory abducens nucleus innervate the retractor bulbi muscles via the VIth nerve. All neurons in the accessory abducens nucleus innervate the retractor bulbi muscles, but gross dissection revealed that the retractor bulbi is also innervated by the IIIrd nerve. The bases for differences between our data and previously published descriptions are discussed. The trochlear nucleus of the rabbit has not been previously studied by methods of axonal transport. The body of the nucleus, its caudal tail, the trajectories of axons entering the trochlear nerve, and soma size distributions are described. The trochlear nucleus contains approximately 900 neurons; most are motoneurons the axons of which travel in the trochlear nerve and decussate in the anterior medullary velum. Approximately 3% of trochlear motor neurons innervate the ipsilateral superior oblique muscle. Their soma size is significantly smaller than that of contralaterally projecting neurons. For comparative purposes, the innervation of extraocular muscles by the trochlear nerve was also investigated in several rodents and carnivores. In all animals studied, the percentage of trochlear neurons innervating the ipsilateral superior oblique muscle was strikingly uniform (2-4%). Gross dissection of the extraocular muscles revealed in the rabbit a muscle, innervated by the trochlear nerve, for which we propose the name "tensor trochleae." In the rabbit, this muscle is innervated by approximately one-third of the trochlear motor neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Abducens Nerve↗

Auditory hair cell innervational patterns in lizards.

The pattern of afferent and efferent innervation of two to four unidirectional (UHC) and two to nine bidirectional (BHC) hair cells of five different types of lizard auditory papillae was determined by reconstruction of serial TEM sections. The species studies were Crotaphytus wislizeni (iguanid), Podarcis (Lacerta) sicula and P. muralis (lacertids), Ameiva ameiva (teiid), Coleonyx variegatus (gekkonid), and Mabuya multifasciata (scincid). The main object was to determine in which species and in which hair cell types the nerve fibers were innervating only one (exclusive innervation), or two or more hair cells (nonexclusive innervation); how many nerve fibers were supplying each hair cell; how many synapses were made by the innervating fibers; and the total number of synapses on each hair cell. In the species studies, efferent innervation was limited to the UHC, and except for the iguanid, C. wislizeni, it was nonexclusive, each fiber supplying two or more hair cells. Afferent innervation varied both with the species and the hair cell types. In Crotaphytus, both the UHC and the BHC were exclusively innervated. In Podarcis and Ameiva, the UHC were innervated exclusively by some fibers but nonexclusively by others (mixed pattern). In Coleonyx, the UHC were exclusively innervated but the BHC were nonexclusively innervated. In Mabuya, both the UHC and BHC were nonexclusively innervated. The number of afferent nerve fibers and the number of afferent synapses were always larger in the UHC than in the BHC. In Ameiva, Podarcis, and Mabuya, groups of bidirectionally oriented hair cells occur in regions of cytologically distinct UHC, and in Ameiva, unidirectionally oriented hair cells occur in cytologically distinct BHC regions.

Afferent Pathways↗

De novo neuromuscular junction formation on human muscle fibres cultured in monolayer and innervated by foetal rat spinal cord: ultrastructural and ultrastructural--cytochemical studies.

Ultrastructural features of neuromuscular junction formation and transverse tubule development were studied utilizing a newly developed model in which human muscle fibres cultured in monolayer are innervated by foetal rat spinal cord with dorsal root ganglia attached. At early innervation (7-10 days), when distinct 'boutons' are contacting muscle fibres, the contacts of nerve terminals with the muscle fibres are, ultrastructurally, superficial and unorganized, and there is no basal lamina-like material between nerve terminals and muscle fibres. A bouton consists, ultrastructurally, of a cluster of small nerve terminals contacting the muscle fibre. At 2-3 weeks of innervation, shallow 'beds' are formed on the muscle fibre just beneath nerve terminals, and occasionally there are irregular and miniscule fragments of basal lamina-like material in the cleft. There is no Schwann cell apposing the nerve terminal at this stage of innervation. After 4-5 weeks of innervation there is more definite basal lamina material in the cleft and suggestive postsynaptic plasmalemmal densities and invaginations. However, there is no Schwann cell apposing the nerve terminal at this stage. At 6-8 weeks of innervation, deep postsynaptic folds are present, a Schwann cell apposes the nerve terminal, and basal lamina surrounds the entire muscle fibre. At all four stages of innervation examined, ultrastructural cytochemistry of alpha-bungarotoxin binding reveals that nicotinic ACh receptors are located exclusively at the neuromuscular junctions. After 1-2 weeks of innervation, very few lanthanum-positive transverse tubules are observed and only in close proximity to the surface membrane. After 3 weeks of innervation, more lanthanum-positive tubules are present, and they are located deeper within the muscle fibre. Five weeks after innervation, somewhat more elaborated tubules (but no lateral sacs) appear, and honeycomb structures are often present. After 6-7 weeks of innervation the tubular system is very elaborate and lateral sacs are present. Hence, this study describes consecutive stages of the formation of neuromuscular junctions and transverse tubules in innervated cultured human muscle, and provides an important basis to which similar studies related to the diseased human muscle can be compared.

Animals↗

Loss of polyneuronal innervation and establishment of a topographical map in the glutaeus muscle of Bufo marinus during generation of secondary muscle cells.

The development of synaptic connections to the toad (Bufo marinus) glutaeus magnus from segmental nerves 8 (N8) and 9 (N9) was determined in the postmetamorphic period. Three different-size toads were studied: small (0.3-2.0 g), medium-size (5-15 g) and large (greater than 20 g). The number of cells in the glutaeus increased about 9-fold during development; this involved the appearance and subsequent maturation of secondary fibres throughout the muscle. The glutaeus in small toads, which consisted almost entirely of primary fibres, was innervated to a similar extent by N8 and N9 as assessed by tetanic contraction measurements. During late development there was a progressive increase in the percentage of the muscle innervated by N9 and a decrease in the percentage innervated by N8. This change in the segmental innervation was accompanied by changes in the innervation of the ventral glutaeus as assessed by intracellular recording. In small toads this surface of the muscle was innervated predominantly by N8, with N9 frequently appearing as a low-efficacy terminal on dually innervated fibres. With further development there was a progressive reduction in the percentage of dually innervated fibres and a concomitant decrease in the percentage innervation of the entire ventral glutaeus by N8. These results suggest that the topographical projection is established by the initial distribution of N9 terminals on the primary fibres of the muscle. The multiple innervation of newly generated fibres and the on-going process of terminal elimination results in N9 terminals, many of which were initially weak, preserving their position in the muscle. This occurs at the expense of N8 terminals, whose relative incidence declines during development. The competitive advantage of N9 motoneurones may be due to their greater capacity to lay down axon collaterals and preferentially innervate newly generated fibres; alternatively N9 terminals may displace N8 terminals, which were initially more efficacious, from dually innervated fibres. Secondary muscle fibres generated throughout the muscle are thus incorporated into an increasingly precise topographical map.

Animals↗

Organization of geniculate and trigeminal ganglion cells innervating single fungiform taste papillae: a study with tetramethylrhodamine dextran amine labeling.

Single gustatory nerve fibers branch and innervate several taste buds. In turn, individual taste buds may receive innervation from numerous gustatory nerve fibers. To evaluate the pattern of sensory innervation of fungiform papilla-bearing taste buds, we used iontophoretic fluorescent injection to retrogradely label the fibers that innervate single taste papillae in the hamster. For each animal, a single taste papilla was injected through the gemmal pore with 3.3% tetramethylrhodamine dextran amine. Fungiform papillae either at the tongue tip (0.5-1.5 mm from the tip) or more posteriorly (1.5-3.0 mm from the tip) were injected. After one to seven days survival, the geniculate and trigeminal ganglia and the tongue were sectioned and examined for labeled cells and fibers, respectively. Analysis of the number and topographic distribution of geniculate cells innervating single taste papillae, shows that: (i) 15 +/- 4 (S.D.) ganglion cells converge to innervate a single fungiform taste bud; (ii) more ganglion cells innervate anterior- (range: 13-35 cells) than posterior-lying buds (range: five to 12 cells), which, in part, may be related to bud volume (microm3); and (iii) ganglion somata innervating a single taste bud are scattered widely within the geniculate ganglion. Analysis of labeled fibers in the tongue demonstrated that two to eight taste buds located within 2 mm of the injected taste bud share collateral innervation with the injected taste bud. Since all buds with labeled fibers were located in close proximity (within a 2-mm radius), widely dispersed geniculate ganglion cells converge to innervate closely spaced fungiform taste buds. Trigeminal ganglion (mandibular division) cells were also labeled in every case and, as with the geniculate ganglion, a dispersed cell body location and collateralization pattern among papillae were observed. This study shows that iontophoresis of tetramethylrhodamine dextran amine, selectively applied to individual peripheral receptor end-organs, effectively locates sensory ganglion cells in two different ganglia that project to these sites. Moreover, the marker demonstrates collateral branches of sensory afferents associated with the labeled fibers and the nearby receptor areas innervated by these collaterals. The labeling of single or clusters of receptor cells, as well as identified sensory afferents, affords future possibilities for combining this technique with immunocytochemistry to establish the relationships of innervation patterns with neurotransmitters and neurotropic substances within identified cells.

Afferent Pathways↗

Specific innervation of guinea-pig superior cervical ganglion cells by preganglionic fibres arising from different levels of the spinal cord.

1. The synaptic contribution of preganglionic nerve fibres arising from the last cervical (C8) and the first seven thoracic spinal cord segments (T1-T7) to neurones of the guinea-pig superior cervical ganglion has been studied by means of intracellular recording during ventral root stimulation in vitro. 2. The majority of neurones received innervation from the middle segments (T2 and T3) of the length of spinal cord from which preganglionic fibres derive; an intermediate number of ganglion cells were innervated by fibres from the segments adjacent to these (T1, T4, and T5), and relatively few neurones by fibres from the most rostral and caudal segments supplying innervation to the ganglion (C8, T6 and T7). 3. Each neurone received preganglionic terminals from multiple thoracic segments (range 1-7, mean = 4-0). The estimated minimum number of preganglionic fibres contacting each neurone was 10, on average. 4. As a rule, the spinal segments innervating a neurone were contiguous. Thus we rarely encountered neurones innervated by segments located both rostrally and caudally to a segment which failed to provide innervation. 5. Neurones tended to be innervated predominantly by axons arising from a single spinal segment, with adjacent segments contributing a synaptic influence that diminished as a function of their distance from the dominant segment. All segments provided dominant innervation to at least some neurones. 6. Stimulating the ventral roots of C8-T7 in vivo showed that the axons arising from each segment produced a characteristic pattern of peripheral effects. Thus different populations of neurones in the superior cervical ganglion of the guinea-pig are innervated by preganglionic axons from different levels of the spinal cord, as originally suggested by Langley (1892) for the cat, dog, and rabbit. 7. On the basis of our in vitro studies we conclude that underlying the specificity of innervation of neurones of the superior cervical ganglion that can be inferred from in vivo experiments is a tendency for individual neurones to be innervated in a systematically graded fashion by a contiguous subset of the eight spinal segments which provide innervation to the ganglion.

Action Potentials↗

The organization and development of compartmentalized innervation in rat extensor digitorum longus muscle.

1. We have examined the innervation of the rat extensor digitorum longus (EDL) muscle by the two extramuscular branches formed from the bifurcation of its muscle nerve. Observations of muscle contractions, recordings of end-plate potentials, and glycogen depletion of young adult muscles show that each branch innervates a separate region or 'compartment' in the muscle. The branch entering the muscle nearer the knee (the K branch) innervates fibres in the anteromedial half of the muscle whereas the branch entering closer to the foot (the F branch) innervates fibres located posterolaterally. Individual EDL motoneurones project either into the K or the F branch and therefore innervate fibres located in one compartment. The boundary between the compartments is usually sharply delineated. No obvious anatomical feature exists within the muscle which would explain the division of the muscle into two distinct regions. 2. The presence of a segmentotopic projection from the spinal cord to the muscle was investigated to evaluate its possible contribution to the compartmental pattern. The most posterior neurones of the EDL motor pool were found to project more frequently to the posterolateral F compartment; similarly, the most anterior neurones most frequently project to the anteromedial K compartment. However, each compartment is innervated by both anteriorly and posteriorly located motoneurones. The segmentotopic projection is too weak to explain the presence of neuromuscular compartments. 3. The post-natal period of synapse elimination appears to play at best a minor role in setting up the compartmentalized innervation. Glycogen depletion and intracellular recording in 1-2-day-old muscles show that each nerve branch innervates fibres in the same region of the muscle as in the adult. Most of the fibres in each compartment are polyneuronally innervated by axons in their own particular nerve branch, although fibres located near the boundary between the two compartments are innervated by axons from both nerve branches. This convergent innervation from the two branches disappears in concert with the elimination of polyneuronal innervation throughout the muscle. A random elimination of these convergent inputs appears adequate to explain the final compartmental pattern. 4. Our findings suggest that the compartmental pattern is primarily the consequence of te segregation of EDL motoneurones into two nerve branches which are directed into separate regions of the muscle.

Animals↗

Development of functional motor innervation in supernumerary hindlimbs of the chick embryo.

1. The development of functional motor innervation in supernumerary chick hindlimbs transplated to host embryos at stages 16--18, was studied throughout the embryonic period from stage 27 when peripheral synapses were first formed. 2. The innervation pattern, defined as the number and sequential order of spinal nerves innervating individual muscles, were determined for supernumerary hindlimbs innervated by thoracic, lumbosacral-thoracic, or wholly lumbosacral spinal cord segments. Spinal nerves were electrically stimulated and muscle contraction was scored visually or by tension measurement and compound action-potential recordings were made from muscle nerves. 3. There was no tendency for spinal nerves to grow to or to synapse with the muscles which they normally innervate. Rather, they formed functional connections with inappropriate muscles, which were maintained throughout the developmental period studied. The localization of the motoneuron cell bodies was confirmed with retrograde transport of horseradish peroxidase (HRP) and shown to be adjacent to the spinal nerves through which their axons exited. Motoneurons innervating specific muscles occupied a similar medial-to-lateral position in the cord in both control and supernumerary limbs. 4. In all supernumerary limbs a definite peripheral innervation pattern was formed, which approximated the normal pattern. Specifically, the relative craniocaudal position of motor columns innervating individual muscles was conserved, even though the actual spinal nerves innervating the transplant were different from animal to animal. In limbs reversed along the anterior-posterior axis, the innervation pattern was also reversed. 5. The results suggest that motoneurons have not been rigidly specified to innervate certain muscles at the time when the limb buds were transplanted. Further, the limb itself can apparently influence the innervation pattern in an important manner, possibly by specifying motoneurons or by subsequently directing axon outgrowth and synapse formation.

Action Potentials↗

Initial innervation of embryonic rat tongue and developing taste papillae: nerves follow distinctive and spatially restricted pathways.

The rat tongue has an extensive, complex innervation from four cranial nerves. However, the precise developmental time course and spatial routes of these nerves into the embryonic tongue are not known, although this knowledge is crucial for studying mechanisms that regulate development and innervation of the lingual taste organs, gustatory papillae and resident taste buds. We determined the initial spatial course of nerves in the developing tongue and papillae, and tested the hypothesis that sensory nerves first innervate the tongue homogeneously and then retract to more densely innervate papillae and taste buds. Antibodies to GAP-43 and neurofilaments were used to label nerve fibers in rat embryo heads from gestational day 11 through 16 (E11-E16). Serial sagittal sections were traced and reconstructed to follow paths of each nerve. In E11 rat, geniculate, trigeminal and petrosal ganglia were labeled and fibers left the ganglia and extended toward respective branchial arches. At E13 when the developing tongue is still a set of tissue swellings, the combined chorda/lingual, hypoglossal and petrosal nerves approached the lingual swellings from separate positions. Only the chorda/lingual entered the tongue base at this stage. At E14 and E15, the well-developed tongue was innervated by all four cranial nerves. However, the nerves maintained distinctive entry points and relatively restricted mesenchymal territories within the tongue, and did not follow one another in common early pathways. Furthermore, the chorda/lingual and glossopharyngeal nerves did not set up an obvious prepattern for gustatory papilla development, but rather seemed attracted to developing papillae which became very densely innervated compared to surrounding epithelium at E15. To effect this dense papilla innervation, sensory nerves did not first innervate the tongue in a homogeneous manner with subsequent retraction and/or extensive redirection of fibers into the taste organs. Results contribute to a set of working principles for development of tongue innervation. Points of entry and initial neural pathways are restricted from time of tongue formation through morphogenesis, suggesting distinctive lingual territories for each nerve. Thus, sensory and motor nerves distribute independently of each other, and sensory innervation to anterior and posterior tongue remains discrete. For taste organ innervation, gustatory papillae are not induced by a prepatterned nerve distribution. In fact, papillae might attract dense sensory innervation because neither chorda/lingual nor glossopharyngeal nerve grows homogeneously to the lingual epithelium and then redistributes to individual papillae.

Animals↗

Overexpression of nerve growth factor in skin causes preferential increases among innervation to specific sensory targets.

The impact of increased levels of skin-derived nerve growth factor (NGF) neurotrophin on sensory and sympathetic innervation to the mouse mystacial pad and postero-orbital vibrissae was determined. Consistent with an approximate doubling of neuron number in trigeminal and superior cervical ganglia, many components of the sensory and sympathetic innervation were substantially enhanced. Although the increased number of neurons raised the possibility that all types of innervation were increased, immunohistochemical analysis indicated that enhanced NGF production had a differential effect upon sensory innervation, primarily increasing unmyelinated innervation. This increased innervation occurred in specific locations known to be innervated by small, unmyelinated fibers, suggesting that NGF modulated sensory innervation density, but not targeting. In contrast, sympathetic innervation was not only increased but also was distributed to some aberrant locations. In the intervibrissal fur of the mystacial pad, both the number of sensory axons and branches appeared increased, whereas in vibrissal follicle sinus complexes, only branching increased. In some areas, sensory ending density was lower than expected based upon the size of the source nerve bundles suggesting that many axons and branches were surviving but failing to form functional endings. Furthermore, the immunochemical profile of innervation was altered in some sensory populations as demonstrated by the coexistence of RT-97 neurofilament labeling in calcitonin gene-related peptide (CGRP) positive axons, by the loss of substance P colocalization in some CGRP axons, and by an absence of neuropeptide Y labeling in tyrosine hydroxylase positive sympathetic axons. Collectively, these results indicate that the NGF mediated increase in neuron number may be selective for particular sets of innervation and that increases among some populations may result from phenotypic switching.

Aging↗

Postnatal changes in cell body size and oxidative enzyme activity of spinal motoneurons innervating the rat tibialis anterior muscle.

The cell body size and succinate dehydrogenase (SDH) activity of spinal motoneurons innervating the superficial and deep regions of the tibialis anterior muscle were studied in rats ranging in postnatal age from 3 to 11 weeks, by retrograde neuronal labeling using fluorescent neuronal tracers. The motoneurons innervating the tibialis anterior muscle were located primarily at the L4 spinal cord segment and those innervating the superficial and deep regions of the muscle were distributed throughout the entire extent of the motoneuron pool. The distribution of the motoneurons during postnatal development was similar to that observed in the adult animal. The mean cell body size of the motoneurons innervating the superficial region of the muscle in rats from 5 to 11 weeks of age was greater than that innervating the deep region at corresponding ages. The mean SDH activity of the motoneurons innervating the deep region of the muscle increased during postnatal development, while there were no changes in the mean SDH activity of those innervating the superficial region during this period. At 11 weeks of age, the motoneurons innervating the deep region of the muscle had a higher mean SDH activity than those innervating the superficial region. An inverse relationship between cell body size and SDH activity of motoneurons innervating both the superficial and deep regions of the muscle was observed, independent of age. These results indicate that motoneurons innervating the superficial and deep regions of the rat tibialis anterior muscle have different developmental patterns with regard to cell body size and SDH activity.

Aging↗