Positional cues governing cell migration in leech neurogenesis.

The stereotyped distribution of identified neurons and glial cells in the leech nervous system is the product of stereotyped cell migrations and rearrangements during embryogenesis. To examine the dependence of long-distance cell migrations on positional cues provided by other tissues, embryos of Theromyzon rude were examined for the effects of selective ablation of various embryonic cell lines on the migration and final distribution of neural and glial precursor cells descended from the bilaterally paired ectodermal cell lines designated q bandlets. The results suggest that neither the commitment of q-bandlet cells to migrate nor the general lateral-to-medial direction of their migration depend on interactions with any other cell line. However, the ability of the migrating cells to follow their normal pathways and to find their normal destinations does depend on interactions with cells of the mesodermal cell line, which appears to provide positional cues that specify the migration pathways.

Leech neurogenesis. I. Positional commitment of neural precursor cells.

This paper reports analyses of the differentiation and distribution of identified peripheral neurons and central 5-HT-containing neurons in embryos of the glossiphoniid leech Theromyzon rude that have been deprived of one of the bilaterally paired major ectodermal cell lines called the n bandlets. Cells descended from a lone surviving n bandlet were abnormally distributed across both sides of the ventral midline. Nevertheless, they produced the complement of identified neurons that they would have produced in a normal embryo. Neurons produced by cells that crossed the midline occupied the normal positions of their absent homologs, as demonstrated by morphometric analysis of normal and n-bandlet-deprived ganglia. Ablations of ectodermal cell lines other than the n bandlets (o and p, or q) allowed the formation of normal distributions of neurons descended from the n bandlets. These results are interpreted as showing that neural precursor cells are committed to occupy particular positions before reaching those positions and probably use positional cues of predominantly nonectodermal origin to recognize those positions. Together, the results reported here and in the accompanying paper (S. Torrence, M. Law, and D. Stuart, 1989, Dev. Biol. 136, 40-60) suggest that ectodermal cells that are committed to give rise to specific neurons use cues provided by the mesoderm to find positions appropriate to their fates.

Leech neurogenesis. II. Mesodermal control of neuronal patterns.

This paper reports analyses of the effects of eliminating mesoderm from one or both sides of embryos of the glossiphoniid leech Theromyzon rude on the differentiation and distribution of ectodermal cells, especially identified peripheral neurons and central 5-hydroxytryptamine (5-HT)-containing neurons arising from the bilateral pair of cell lines called the n bandlets (n-kinship cells). In mesoderm-deprived regions, no segmental hemiganglia formed, and identified neurons were not organized into recognizable patterns, although 5-HT neurons underwent neurochemical differentiation and grew axons. In unilaterally mesoderm-deprived embryos, segmental hemiganglia were formed in a midbody experimental zone, and cells that had abnormally crossed the ventral midline from the deprived side gave rise to identified neurons that were incorporated as supernumeraries into the normal organization of hemiganglia on the nondeprived side. In a posterior experimental zone, ganglionic morphology was disrupted on both sides. We conclude that precursor cells are committed to specific neuronal fates regardless of whether they occupy normal positions and that mesodermal tissues provide positional cues necessary for such precursor cells to find positions appropriate to their fates.

Gangliogenesis in leech embryos: migration of neural precursor cells.

In the metameric CNS of leeches, identified neurons occupy highly stereotyped positions in each segmental ganglion. Although many of the neural precursor cells arise near their definitive positions, some arise outside the prospective domain of the segmental ganglia and thus must migrate into the CNS. Here, we report the results of an analysis of the role of cell migration in gangliogenesis in the leech Theromyzon rude. Segmental ganglia of the ventral nerve cord arise as laterally thickened sheets of tissue lying astride the ventral midline. Particular identified circular and longitudinal muscle fibers, visualized by indirect immunofluorescence using a monoclonal antibody against leech muscle, outline the presumptive ganglionic territories even before the ganglionic rudiments become morphologically distinct and serve as anatomical landmarks to which the cell movements are related. Cell lineage tracers microinjected into precursor blastomeres are used to visualize migratory cells. Small groups of neural precursor cells that arise outside the prospective ganglionic territories migrate with stereotyped timing along stereotyped pathways to reach their definitive positions, and each group of migratory cells gives rise to a stereotyped subset of the cells in a ganglion. No segmental or regional differences are observed in any aspect of cell migration studied here, supporting the view that segmental differences in the architecture of the leech CNS arise only after the initial condensation of the ganglionic rudiments.

Rhythmic contractions of the ampullar epidermis during metamorphosis of the ascidian Molgula occidentalis.

The ampullae of Molgula occidentalis are hollow, tubular extensions of the epidermis. They are ensheathed by a secreted tunic. When they grow out shortly after settlement, the ampullae spread the tunic over the substratum to form a firm attachment for the sessile juvenile. A simple squamous epithelium forms the thin ampullar walls. A glandular, simple columnar epithelium forms the distal tip of each ampulla. The glandular cells probably secrete the adhesive that attaches the tunic to the substratum. Repetitive, peristaltic contractions pass from the base to the distal end of each ampulla. Microsurgery, time-lapse cinemicrography and TEM have been used to analyze this phenomenon. The contractions are mediated by a layer of 4-8 nm microfilaments in the base of the ampullar epithelium. Each juvenile has 7-9 ampullae which contract at different frequencies. Isolated ampullae continue to contract normally for several days. Thus each ampulla has an intrinsic rhythm. Microsurgical experiments suggest that there is no specific region within an ampulla with unique pacemaker properties. It is proposed that communication via gap junctions allows the coordination of ampullar cells into a well organized peristaltic wave.