Sacral neural crest cell migration to the gut is dependent upon the migratory environment and not cell-autonomous migratory properties.

Avian neural crest cells from the vagal (somite level 1-7) and the sacral (somite level 28 and posterior) axial levels migrate into the gut and differentiate into the neurons and glial cells of the enteric nervous system. Neural crest cells that emigrate from the cervical and thoracic levels stop short of the dorsal mesentery and do not enter the gut. In this study we tested the hypothesis that neural crest cells derived from the sacral level have cell-autonomous migratory properties that allow them to reach and invade the gut mesenchyme. We heterotopically grafted neural crest cells from the sacral axial level to the thoracic level and vice versa and observed that the neural crest cells behaved according to their new position, rather than their site of origin. Our results show that the environment at the sacral level is sufficient to allow neural crest cells from other axial levels to enter the mesentery and gut mesenchyme. Our study further suggests that at least two environmental conditions at the sacral level enhance ventral migration. First, sacral neural crest cells take a ventral rather than a medial-to-lateral path through the somites and consequently arrive near the gut mesenchyme many hours earlier than their counterparts at the thoracic level. Our experimental evidence reveals only a narrow window of opportunity to invade the mesenchyme of the mesentery and the gut, so that earlier arrival assures the sacral neural crest of gaining access to the gut. Second, the gut endoderm is more dorsally situated at the sacral level than at the thoracic level. Thus, sacral neural crest cells take a more direct path to the gut than the thoracic neural crest, and also their target is closer to the site from which they initiate migration. In addition, there appears to be a barrier to migration at the thoracic level that prevents neural crest cells at that axial level from migrating ventral to the dorsal aorta and into the mesentery, which is the portal to the gut.

Dynein-dynactin function and sensory axon growth during Drosophila metamorphosis: A role for retrograde motors.

Mutations in the genes for components of the dynein-dynactin complex disrupt axon path finding and synaptogenesis during metamorphosis in the Drosophila central nervous system. In order to better understand the functions of this retrograde motor in nervous system assembly, we analyzed the path finding and arborization of sensory axons during metamorphosis in wild-type and mutant backgrounds. In wild-type specimens the sensory axons first reach the CNS 6-12 h after puparium formation and elaborate their terminal arborizations over the next 48 h. In Glued1 and Cytoplasmic dynein light chain mutants, proprioceptive and tactile axons arrive at the CNS on time but exhibit defects in terminal arborizations that increase in severity up to 48 h after puparium formation. The results show that axon growth occurs on schedule in these mutants but the final process of terminal branching, synaptogenesis, and stabilization of these sensory axons requires the dynein-dynactin complex. Since this complex functions as a retrograde motor, we suggest that a retrograde signal needs to be transported to the nucleus for the proper termination of some sensory neurons.

Development of the spinal nerves in the mouse with special reference to innervation of the axial musculature.

Development of the mouse spinal nerves was studied. On E11 (11th day of gestation), the primitive spinal nerve fascicle extended ventrally in the anterior half of the sclerotome. Spinal nerves in the forelimb region united with each other to form the primitive brachial plexus. Their terminal segment was covered by a peculiar cell mass. On E12, five primary branches developed along the primitive spinal nerve trunk. The ramus dorsalis was originally a cutaneous nerve, supplying two series of branches to the skin of the back. The medial series was derived from the dorsal ramus of C2-C8, and the lateral series from C8 and the more caudal dorsal rami. Nerves of the former series took the presegmental course through the intermyotomic space, while those of the latter the postsegmental course. The ramus cutaneous lateralis was a nerve that took the presegmental course to become cutaneous. The ramus intercostalis externus was a muscle branch whose distribution was restricted within the segment. The ramus anterior was a muscle branch from the end of the primitive spinal nerve trunk. The ramus visceralis connected a thoracic nerve with the para-aortic sympathetic cell cord. On E13-16 the ramus anterior secondarily gave off a cutaneous branch (ramus cutaneous anterior). The ramus intercostalis externus extended ventrally deep to the intercostalis externus muscle, crossing just caudal to the ramus cutaneous lateralis that secondarily gave off branches to the obliquus externus abdominis muscle.

Alteration of pectoral fin nerves following ablation of fin buds and by ectopic fin buds in the Japanese medaka fish.

The role of the pectoral fin bud for outgrowth by fin axons was assessed by ablation of pectoral fin buds and by transplantation of fin buds to ectopic sites in the embryos of the Japanese medaka fish (Oryzias latipes). Normally nerves from segments 1-4 (S1-4) and less frequently the S5 nerve converged at the base of the fin bud by extending toward the fin bud on the ventral surface of the axial muscles (H. Okamoto and J. Y. Kuwada, 1991, Dev. Biol. 146). Following ablation of the fin bud before motor growth cones have begun to extend laterally, nerves in S1-5 followed a trajectory down the middle of each segment parallel to the borders of the metamerically arranged axial muscles rather than converging. This trajectory was similar to that of more posterior segmental nerves which do not converge toward the fin bud. When fin buds were transplanted to more posterior segments, nerves from S1-5 often changed their trajectories and extended to the base of ectopic buds. Furthermore, motor nerves from segments posterior to S5, which normally do not innervate the fin bud, also extended to the ectopic fin bud. When faced with both the host and ectopic fin bud, motor nerves extended to either fin bud or branched and extended to both fin buds. These results demonstrate that the early fin bud is necessary for correct outgrowth of fin nerves and suggest that the fin bud normally attracts fin nerves to its base. One possible mechanism for the attraction of motor growth cones by the fin bud is a long distance cue emitted by the fin bud.

The development of GABA-like immunoreactivity in the thoracic ganglia of the locust Schistocerca gregaria.

The development of GABA-like immunoreactivity was investigated in embryonic and juvenile locusts using an antibody raised against GABA-protein conjugates. GABA-like immunoreactivity was first detectable in the neuropile of embryonic ganglia at 55% development, and in neuronal somata at 62% development. The total number of immunoreactive somata increased between 62% and 85% embryonic development, and followed an anterio-posterior pattern of expression. At 85% development, the number of immunoreactive somata reached adult levels and no change in number was then seen. In embryonic stages and first and second juvenile instars two dorsal and four ventral groups of somata were labeled in all three thoracic ganglia, whilst in later juvenile instars one of the dorsal groups was visible as a separate entity only in the metathoracic ganglion. These early patterns were modified by alterations in the positions of some of the groups during late embryogenesis and during juvenile development to produce the adult pattern. The results show that the development of GABA expression is similar to that of other neurotransmitters. The characteristics of the development of immunoreactivity indicate that some of these immunoreactive clusters may be derived from clonally related neurones. Finally, we demonstrate the presence of immunoreactive somata and processes in embryos, which correspond to those of identified local and intersegmental interneurones studied in the adult.

Brachial muscles of dystrophic chick embryos atypically sustain interaction with thoracic nerves.

Previous analyses of experimental chick embryos of normal lineage demonstrate the inability of brachial muscles to sustain a successful union with foreign nerves derived from a thoracic neural tube segment transplanted to the brachial region at day 2 in ovo (day 2E). The present experiments were performed to determine if mutant chick embryos afflicted with hereditary muscular dystrophy would respond similarly to this experimental manipulation. Using the same criteria applied to our analysis of experimental normal embryos, our results demonstrated that dystrophic brachial muscles were capable of maintaining a compatible union with foreign thoracic nerves throughout the experimental period analysed. Significant muscle growth occurred, intramuscular nerve branches were maintained, motor endplates formed and wing motility was equivalent to that of unoperated dystrophic embryos. Thus, foreign nerves rejected by normal brachial muscles were accepted by brachial muscles of the mutant dystrophic embryo.

A method for fate mapping the foci of lethal and behavioral mutants in Drosophila melanogaster.

A new method of mosaic fate mapping, called focusing, is introduced. Its advantages are that it allows a mapping, on the blastoderm surface, of the site of action of functions defined by either pre-adult lethal or behavioral mutations. Moreover, it does not require that the mosaics used be 50% of one genotype and 50% of the other. Methods for quantitative evaluation of the results of focusing, and a comparison of this method with others, are discussed.--Focusing is applied to the analysis of a new mutant, doomed (symbol: dmd), a distally located X-linked recessive in D. melanogaster, that causes the deaths of males and females around the time of eclosion. The dmd phenotype among eclosing flies is first seen as the loss of thoracic motor coordination and as ether sensitivity. Fate mapping by the method of focusing places the site of action of the dmd+ function in the same region of the map as that of the thoracic neural ganglia primordia, but not that of muscle, suggesting the possibility that the effect of dmd is on a thoracic neural function rather than a muscular one.