Biomedical subjects

S E Fraser

Publications and source records attributed to S E Fraser.

At least 91 records · Page 5Linked to original sources

PubMed1990

Dynamic changes in optic fiber terminal arbors lead to retinotopic map formation: an in vivo confocal microscopic study.

Dynamic remodeling of retinal ganglion cell terminal arbors has been proposed to contribute to formation of the topographically ordered retinotectal projection. To test this directly, the growth of individual terminal arbors was observed in live X. laevis tadpoles using a confocal microscope to visualize their complex three-dimensional structure. During initial development, nasal and temporal retinal arbors covered overlapping tectal areas. Despite subsequent remodeling, the dimensions and positions of the temporal arbors remained relatively stable. In contrast, the nasal arbors grew caudally, as they extended caudal branches and retracted rostral branches. These results suggest that differences in the remodeling of the nasal and temporal arbors lead to the emergence of retino-topography along the rostrocaudal axis of the tectum. All the terminal arbors were dynamic, including those with stable dimensions, suggesting that continual remodeling of arbors may be a universal feature of neuronal projections.

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PubMed1990

Acetylcholine receptor clustering is triggered by a change in the density of a nonreceptor molecule.

Acetylcholine receptors become clustered at the neuromuscular junction during synaptogenesis, at least in part via lateral migration of diffusely expressed receptors. We have shown previously that electric fields initiate a specific receptor clustering event which is dependent on lateral migration in aneural muscle cell cultures (Stollberg, J., and S. E. Fraser. 1988. J. Cell Biol. 107:1397-1408). Subsequent work with this model system ruled out the possibility that the clustering event was triggered by increasing the receptor density beyond a critical threshold (Stollberg, J., and S. E. Fraser. 1990. J. Neurosci. 10:247-255). This leaves two possibilities: the clustering event could be triggered by the field-induced change in the density of some other molecule, or by a membrane voltage-sensitive mechanism (e.g., a voltage-gated calcium signal). Electromigration is a slow, linear process, while voltage-sensitive mechanisms respond in a rapid, nonlinear fashion. Because of this the two possibilities make different predictions about receptor clustering behavior in response to pulsed or alternating electric fields. In the present work we have studied subcellular calcium distributions, as well as receptor clustering, in response to such fields. Subcellular calcium distributions were quantified and found to be consistent with the predicted nonlinear response. Receptor clustering, however, behaves in accordance with the predictions of a linear response, consistent with the electromigration hypothesis. The experiments demonstrate that a local increase in calcium, or, more generally, a voltage-sensitive mechanism, is not sufficient and probably not necessary to trigger receptor clustering. Experiments with slowly alternating electric fields confirm the view that the clustering of acetylcholine receptors is initiated by a local change in the density of some non-receptor molecule.

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PubMed1990

Pathways of trunk neural crest cell migration in the mouse embryo as revealed by vital dye labelling.

Analysis of neural crest cell migration in the mouse has been difficult due to the lack of reliable cell markers. Recently, we found that injection of DiI into the chick neural tube marks premigratory neural crest cells whose endfeet are in contact with the lumen of the neural tube (Serbedzija et al. Development 106, 809-819 (1989)). In the present study, this technique was applied to study neural crest cell migratory pathways in the trunk of the mouse embryo. Embryos were removed from the mother between the 8th and the 10th days of development and DiI was injected into the lumen of the neural tube. The embryos were then cultured for 12 to 24 h, and analyzed at the level of the forelimb. We observed two predominant pathways of neural crest cell migration: (1) a ventral pathway through the rostral portion of the somite and (2) a dorsolateral pathway between the dermamyotome and the epidermis. Neural crest cells were observed along the dorsolateral pathway throughout the period of migration. The distribution of labelled cells along the ventral pathway suggested that there were two overlapping phases of migration. An early ventrolateral phase began before E9 and ended by E9.5; this pathway consisted of a stream of cells within the rostral sclerotome, adjacent to the dermamyotome, that extended ventrally to the region of the sympathetic ganglia and the dorsal aorta.(ABSTRACT TRUNCATED AT 250 WORDS)

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PubMed1990

In situ analysis of neuronal dynamics and positional cues in the patterning of nerve connections.

Recently developed imaging techniques permit individual cells to be uniquely labeled and followed over time as development proceeds in intact vertebrate embryos. Small groups of cells in the developing eye rudiment of the frog Xenopus have been labeled with the vital dyes DiI, lysinated fluorescein dextran (LFD) or lysinated rhodamine dextran (LRD). Individual optic axons and their growth cones were clearly visible in the intact living animal using confocal microscopy or epifluorescence microscopy with a low light level video camera and computer-based video image enhancement. To follow the dynamics of single optic nerve fiber terminal arborizations, small groups of cells, or even single retinal ganglion cells, were labeled with DiI, and the resulting labeled optic nerve fibers were imaged using a confocal microscope. The images show a profound alteration in morphology from day to day, demonstrating that optic nerve terminal arborizations are dynamic structures constantly extending and retracting branches. To follow the topography of the developing projection and analyze the cues that guide its formation, small groups of eyebud cells from LFD- and LRD-labeled donor embryos were grafted to an unlabeled host in either a location equivalent to that from which they had been removed (homotopic grafts) or a non-equivalent location (heterotopic grafts). Axons from homotopic grafts projected to the tectum as expected from the adult topography of the retinotectal projection. Dorsoventral topography was present from the time that the optic nerve fibers were observable in the tectum, in agreement with previous work. Nasotemporal topography was subtle or absent for the first few days, and then slowly refined. The importance of positional cues was tested by performing heterotopic eyebud grafts, in which the labeled eyebud cells are grafted to inappropriate places in the host eyebud. The heterotopic grafts appeared to integrate with the ectopic site in the eyebud in a functional manner. They should, therefore, project to the tectum together with their new neighbors if neighbor interactions or activity-based cues were of primary importance in the initial patterning of the map. However, the experiments showed that the axons from heterotopic grafts always behaved in a fashion appropriate to their position of origin in the donor, regardless of their final position in the host. These observations indicate that small groups of eyebud cells (as small as a single cell) possess positional information that plays a dominant role in guiding the optic nerve fibers to their target sites in the tectum.

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PubMed1990

Local accumulation of acetylcholine receptors is neither necessary nor sufficient to induce cluster formation.

Acetylcholine receptors (AChRs) accumulate at developing neuromuscular junctions in part via lateral migration of diffusely expressed receptors. Using a model system--cultured Xenopus muscle cells exposed to electric fields--we have shown that AChRs, concentrated at the cathode-facing cell pole, continue to aggregate there after the field is terminated (Stollberg and Fraser, 1988). These observations are consistent with the possibility that the field-induced increase in receptor concentration triggers the aggregation event. Only 2 other molecular events could initiate the electric field-induced receptor aggregation: (1) a local increase in the density of some other molecules, or (2) a voltage-sensitive mechanism. Treatment of muscle cell cultures with neuraminidase changes the cell surface charge and has been reported to reverse the direction of electromigration for AChRs and concanavalin A binding sites (Orida and Poo, 1978). Using digitally analyzed fluorescence videomicroscopy, we find that AChRs in neuraminidase-treated cultures accumulate at both cell poles in an electric field. After termination of the field, the AChR continues to aggregate at the cathode-facing pole, as in cells not treated with neuraminidase. However, receptor density decreases at the anode-facing pole, indicating that elevated AChR density does not initiate receptor aggregation. Cells pretreated with neuraminidase and trypsin (which blocks receptor aggregation) display reversed receptor distributions compared to untreated controls, indicating that electromigration has indeed been reversed. The rate at which neuraminidase- and trypsin-treated cells approach steady-state distributions indicates a receptor diffusion constant of approximately 1.2 x 10(-9) cm2/sec, consistent with a diffusion trap mechanism of receptor aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)

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PubMed1989

Cell lineage analysis reveals multipotent precursors in the ciliary margin of the frog retina.

The eyes of lower vertebrates grow throughout life by the proliferation of cells in the ciliary margin. To determine the range of cell types that descend from single cells of the ciliary margin in Xenopus laevis embryos, a cell lineage study was performed. Precursor cells in the periphery of the eyes were labeled by intracellular injection of rhodamine dextran, and the proliferation of these cells resulted in clones of labeled descendants. The number of cells per clone was quite variable (range, 1 to 104 cells). This broad range of sizes (compared to clones derived from optic vesicle cells) and their distribution suggest that there may be two types of ciliary margin cells with different proliferative fates: self-renewing stem cells and cells that undergo a limited number of divisions. Labeled descendants in the neural retina differentiated into all of the major cell types, including glia. One-quarter of the clones had labeled descendants in both the neural retina and the pigmented epithelium. These observations suggest that ciliary margin cells are multipotent for all neural, glial, and pigmented epithelial cell types of the eye. Thus, the molecular events that regulate the commitment of specific cell types must occur late in the cell lineages of the ciliary margin.

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PubMed1989

Electrophoretic repatterning of charged cytoplasmic molecules within tissues coupled by gap junctions by externally applied electric fields.

Ionic currents and cytoplasmic voltage gradients have been observed in a variety of polarizing cells and developing tissues. In certain cases, it has been determined that these endogenous electric fields can segregate intracellular charged molecules by electrophoresis; in other cases, the endogenous fields are suspected to have such an influence. Separate theoretical suggestions have been made that extracellular electric currents, whether from a biological or a nonbiological source, should be able to electrophorese intracellular molecules after being conducted through cell membranes into the interior of long single cells [L.F. Jaffe and R. Nuccitelli (1977) Annu. Rev. Biophys. Bioeng. 6, 445-476] or extended ensembles of cells coupled electrotonically by gap junctions [M.S. Cooper (1984) J. Theor. Biol. 111, 123-130]. To test whether external electric fields could redistribute intracellular molecules within a tissue coupled by gap junctions, and to quantitatively measure in situ the electrophoretic mobility of a charged intracellular molecule, we injected 6-carboxyfluorescein into the electrotonically coupled lateral giant neurons of the crayfish abdominal nerve cord. When a dc electric field (0.2-3.4 V/cm) was subsequently applied along the length of the cord, the negatively charged fluorescent dye was observed to migrate through both the cytoplasms and the gap junctions of the lateral giant neurons, toward the anode, at a rate directly proportional to the applied electric field strength (electrophoretic mobility = -0.92 +/- 0.27 micron/sec per V/cm). These results suggest that electric fields of a sufficient magnitude, whether of an exogenous or an endogenous origin, can repattern the distribution of charged molecules within the cytoplasm of an extended ensemble of coupled cells. In addition, these results suggest that externally applied electric fields might be used in studies of pattern formation to repattern the intercellular distribution of charged molecules that are permeant to gap junctions within electrically coupled tissues.

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PubMed1989

Gradual appearance of a regulated retinotectal projection pattern in Xenopus laevis.

The topographic projection pattern formed by the retinal ganglion cell axons in the tectum of the lower vertebrate appears to require positional cues that guide the optic nerve fibers to their appropriate targets. One approach to understanding these positional cues or "positional information" has been to investigate changes in the pattern of the retinotectal projection after surgical manipulation of the embryonic eyebud. Analysis of these apparent changes in the patterns of positional information in the eye, termed "pattern regulation," may provide clues to both the nature of positional information and the mechanisms by which it is assigned to cells in the eyebud. Here we examine pattern regulation in the Xenopus visual system following the replacement of the temporal half of a right eyebud with the temporal half of a left eyebud. This manipulation requires that the left half-eyebud be inverted along its dorsoventral axis. Electrophysiological maps of these compound eyes in postmetamorphic frogs reveal regulated maps; the cells in the temporal half of the NrTl eye project to the tectum with a dorsoventral polarity appropriate for their position in the host eye and not appropriate for the original positions of the grafted cells in the donor eyebud. Paradoxically, the regulated patterns are not apparent in the projections of the original grafted eyebud cells during early larval development. Using fiber-tracing and electrophysiological mapping techniques, we now show that the regulated patterns appear gradually in the projections made by peripheral retinal cells added during mid-larval development. Because the regulation occurs relatively late in development and probably only in the peripheral retinal cells, simple models of epimorphic or morphallactic regulation do not appear to fit this system. Thus, new or more complex models must be invoked to explain the phenomenon of pattern regulation in the developing visual system of Xenopus.

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PubMed1989

Slow intermixing of cells during Xenopus embryogenesis contributes to the consistency of the blastomere fate map.

The relatively consistent fates of the blastomeres of the frog embryo could result from (i) predetermination of the blastomeres or (ii) reproducible morphogenetic cell movements. In some species, the mixing of the cells during development provides a test between these alternative hypotheses. If blastomeres are predetermined, then random intermixing of the descendants with neighboring cells could not alter their fate. To follow cell mixing during Xenopus development, fluorescent dextran lineage tracers were microinjected into identified blastomeres at the 16-cell stage. The labelled descendants of the injected blastomeres were followed over several stages of embryogenesis. After gastrulation, the labelled descendants formed relatively coherent groups in characteristic regions of the embryo. By larval stages, most of the labelled descendants were still located in characteristic regions. However, coherence was less pronounced and individual descendants were located in many regions of the embryo. Hence, cell mixing is a slow, but progressive, process throughout Xenopus development. This is in sharp contrast to the extensive mixing that occurs during the early development of other vertebrates, such as zebrafish and mice. The slow cell mixing in Xenopus development suggests a simple mechanism for the consistent fates of cleavage-stage blastomeres. The stereotyped cell movements of embryogenesis redistribute the largely coherent descendants to characteristic locations in the embryo. The small amount of mixing that does occur would result in variable locations of a small proportion of the descendants; this could contribute to the observed variability of the blastomere fate map. Because cell mixing during Xenopus development is insufficient to challenge possible lineage restrictions, additional experiments must be performed to establish when and if lineage restrictions occur.

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PubMed1989

The oral-aboral axis of a sea urchin embryo is specified by first cleavage.

Several lines of evidence suggest that the oral-aboral axis in Strongylocentrotus purpuratus embryos is specified at or before the 8-cell stage. Were the oral-aboral axis specified independently of the first cleavage plane, then a random association of this plane with the blastomeres of the four embryo quadrants in the oral-aboral plane (viz. oral, aboral, right and left) would be expected. Lineage tracer dye injection into one blastomere at the 2-cell stage and observation of the resultant labeling patterns demonstrates instead a strongly nonrandom association. In at least ninety percent of cases, the progeny of the aboral blastomeres are associated with those of the left lateral blastomeres and the progeny of the oral blastomeres with the right lateral ones, respectively. Thus, ninety percent of the time the oral pole of the future oral-aboral axis lies 45 degrees clockwise from the first cleavage plane as viewed from the animal pole. The nonrandom association of blastomeres after labeling of the 2-cell stage implies that there is a mechanistic relation between axis specification and the positioning of the first cleavage plane.

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PubMed1989

A vital dye analysis of the timing and pathways of avian trunk neural crest cell migration.

To permit a more detailed analysis of neural crest cell migratory pathways in the chick embryo, neural crest cells were labelled with a nondeleterious membrane intercalating vital dye, DiI. All neural tube cells with endfeet in contact with the lumen, including premigratory neural crest cells, were labelled by pressure injecting a solution of DiI into the lumen of the neural tube. When assayed one to three days later, migrating neural crest cells, motor axons, and ventral root cells were the only cells types external to the neural tube labelled with DiI. During the neural crest cell migratory phase, distinctly labelled cells were found along: (1) a dorsolateral pathway, under the epidermis, as well adjacent to and intercalating through the dermamyotome; and (2) a ventral pathway, through the rostral portion of each sclerotome and around the dorsal aorta as described previously. In contrast to those cells migrating through the sclerotome, labelled cells on the dorsolateral pathway were not segmentally arranged along the rostrocaudal axis. DiI-labelled cells were observed in all truncal neural crest derivatives, including subepidermal presumptive pigment cells, dorsal root ganglia, and sympathetic ganglia. By varying the stage at which the injection was performed, neural crest cell emigration at the level of the wing bud was shown to occur from stage 13 through stage 22. In addition, neural crest cells were found to populate their derivatives in a ventral-to-dorsal order, with the latest emigrating cells migrating exclusively along the dorsolateral pathway.

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PubMed1988

Cell lineage analysis reveals multipotency of some avian neural crest cells.

A major question in developmental biology is how precursor cells give rise to diverse sets of differentiated cell types. In most systems, it remains unclear whether the precursors can form many or all cell types (multipotent or totipotent), or only a single cell type (predetermined). The question of cell lineage is central to the neural crest because it gives rise to numerous and diverse derivatives including peripheral neurons, glial and Schwann cells, pigment cells, and cartilage. Although the sets of derivatives arising from different populations of neural crest cells have been well-documented, relatively little is known about the developmental potentials of individual neural crest cells. We have iontophoretically microinjected the vital dye, lysinated rhodamine dextran (LRD) into individual dorsal neural tube cells to mark unambiguously their descendants. Many of the resulting labelled clones consisted of multiple cell types, as judged by both their location and morphology. Cells as diverse as sensory neurons, presumptive pigment cells, ganglionic supportive cells, adrenomedullary cells and neural tube cells were found within individual clones. Our results indicate that at least some neural crest cells are multipotent before their departure from the neural tube.

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PubMed1988

Multipotent precursors can give rise to all major cell types of the frog retina.

A prospective lineage analysis was performed to determine the variety of cell types that could be formed by individual precursor cells of the developing frog retina. Fluorescent dextran was iontophoretically injected into single cells of the embryonic optic vesicle. After further development of the embryo, labeled descendants were observed in all three layers of the larval retina. Furthermore, different clones were composed of various combinations of all major cell types, including the glial Müller cells. Hence, single optic vesicle cells have the potential to form any type of retinal cell, suggesting that the interactions that specify the differentiation pathway of retinal cells must occur late in development.

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PubMed1988

Application of new technologies to studies of neural crest migration and differentiation.

This review describes the application of new techniques for examining some longstanding questions in the neural crest system concerning pathways of migration, cell lineage decisions, and importance of the extracellular matrix. The first issue examined involves the migratory pathways followed by neural crest cells. In birds, it has been possible to map crest migratory routes accurately using antibodies that selectively recognize neural crest cells. These antibodies permit the identification of migrating cells in the absence of the surgical trauma incurred by neural tube transplantations. In amphibian embryos, which are more readily accessible to embryological manipulation than are birds, neural fold grafts using two new cell markers have made it possible to map the early stages of neural crest migration using both interspecific and intraspecific chimerae. In both birds and amphibians, a metameric pattern of neural crest migration was observed; this presumably results from interactions with the adjacent somites. A second novel experimental paradigm tests the role of cell surface-extracellular matrix interactions in neural crest migration. Using antibodies to perturb interactions between the cell surface and the extracellular matrix selectively, it has been possible to identify some molecular interactions that are important for aspects of neural crest migration in situ. New techniques have also made it possible to examine the cell lineage decisions of neural crest cells. Using a retrograde labeling technique and a method of microinjecting cells into embryos, we have found that neural crest-derived cholinergic neurons remain plastic with respect to their neurotransmitter expression even after overt differentiation. New cell-marking techniques have made it possible to study neural crest cell lineage by means of clonal analysis. Individual neural crest precursors can be injected with a cell lineage tracer that is passed to all of its progeny. Thus, we can examine the developmental potential of individual neural crest cells. The various experimental paradigms presented in this review illustrate the utility of recent technological advances to study embryological questions that have long concerned investigators interested in the neural crest and in other developmental systems.

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PubMed1988

Alterations in the Xenopus retinotectal projection by antibodies to Xenopus N-CAM.

The patterned neural projection from the eye to the optic tectum of lower vertebrates (the retinotectal projection) has been proposed to be ordered by interactions between the optic nerve fibers and their surrounding tissues. To investigate the role of one such defined cell interaction, agarose implants containing antibodies to the neural cell adhesion molecule, N-CAM, were inserted into the tectum of the African clawed frog, Xenopus laevis. Both monoclonal and polyclonal antibodies against N-CAM reversibly and specifically distorted the pattern of the retinotectal projection, decreasing the precision of the projection as determined by electrophysiological techniques as well as decreasing the density of retinal innervation of the tectum and the branching of single axons as determined by horseradish peroxidase tracing. The anatomical effects became maximal at 4 to 6 days after implantation and returned to undetectable levels by 2 weeks, whereas the physiological effects became maximal by 8 to 10 days and a normal physiological map was reestablished within 4 weeks. The results are consistent with the hypothesis that anti-N-CAM antibodies perturb the ongoing growth and retraction of the terminal arbors of the optic nerve fibers, such that a region of the tectum becomes largely denuded of fibers. The physiological defects may then be a consequence both of the initial retraction of optic nerve terminals and of the rapid ingrowth of the perturbed and neighboring optic nerve fibers into the denuded region after the antibodies were cleared from the tectum. These results support the concept of a major role for N-CAM-mediated adhesion during map regeneration and maintenance.

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PubMed1988

Mapping of neural crest pathways in Xenopus laevis using inter- and intra-specific cell markers.

This study examines the pathways of migration followed by neural crest cells in Xenopus embryos using two recently described cell marking techniques. The first is an interspecific chimera created by grafting Xenopus borealis cells into Xenopus laevis hosts. The cells of these closely related species can be distinguished by their nuclear dimorphism. The second type of marker is created by microinjection of lysinated dextrans into fertilized eggs which can then be used for intraspecific grafting. These recently developed fluorescent dyes are fixable and identifiable in both living and fixed embryos. After grafting labeled donor neural tubes into unlabeled host embryos, the distribution of neural crest cells at various stages after grafting was used to define the pathways of neural crest migration. To control for possible grafting artifacts, fluorescent lysinated dextran was injected into a single blastomere which gives rise to a large number of neural crest cells, thereby labeling the neural crest without grafting. By all three techniques, Xenopus neural crest cells were observed along two predominant pathways in the trunk. The majority of neural crest cells were observed along a "ventral" route, between the neural tube and somite, the notochord and somite, and along the dorsal mesentery. A second group of neural crest cells was observed "dorsally" where they populated the dorsal fin. A third minor "lateral" pathway was observed primarily in borealis/laevis chimerae and in blastomere-injected embryos; some neural crest cells were observed underneath the ectoderm lateral to the neural tube. Along the rostrocaudal axis, neural crest cells were not continuously distributed but were primarily located across from the caudal two-thirds of the somite. Fewer than 3% of the neural crest cells were observed across from the rostral third of each somite. When grafted to ventral locations, neural crest cells were not able to migrate dorsally but migrated laterally along the dorsal mesentery. Labeled neural crest cells gave rise to cells of the spinal, sympathetic, and enteric ganglia as well as to adrenal chromaffin cells, Schwann cells, pigment cells, mesenchymal cells of the dorsal fin, and some cells in the integuments and in the region of the pronephros. These results show that the neural crest migratory pathways in Xenopus differ from those in the avian embryo. In avians NC cells migrate as a closely associated sheet of cells while in Xenopus they migrate as individual cells. Both species exhibit a metamerism in the neural crest cell distribution pattern along the rostrocaudal axis.(ABSTRACT TRUNCATED AT 400 WORDS)

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PubMed1988

Wound healing, cell communication, and DNA synthesis during imaginal disc regeneration in Drosophila.

Wound healing, gap-junctional cell communication, and DNA synthesis were studied in 3/4 fragments of the imaginal wing disc of Drosophila melanogaster cultured in the female adult abdomen. Such fragments regenerate the missing sector by cell proliferation over a period of several days. Individual cells were marked in the starting fragment by intracellular injection of high-molecular-weight lysinated rhodamine dextran, and the positions of the marked cells were determined by fluorescence microscopy before and after various culture times. These experiments showed that cells along the two wound edges were brought together by wound healing during the first day of the culture period. The marked cells then gradually moved apart over the next 3 days as new cells were added between them by intercalary regeneration. Gap-junctional cell communication across the healed wound, assessed by the movement of intracellularly injected small fluorescent dye molecules, was not detectable after 1 day of culture. After 2 days there was some dye transfer into the growth zone but not as much as in other directions. After 3 days there was significant dye transfer into the growth zone. DNA synthesis, detected by immunolocalization of incorporated bromodeoxyuridine, was first apparent at 18 hr after the beginning of the culture period; at this time as well as at later times it occurred only in the immediate vicinity of the healed wound. DNA synthesis was stimulated over a distance of several cell diameters from the wound, and for one of the cuts it was much more prevalent on one side of the wound than on the other. The results indicate that wound healing brings together cells from distant positions in the disc, and that the response to this is local DNA synthesis and cell proliferation. Gap-junctional communication across the wound does not appear to be necessary for stimulating this cell proliferation, although the timing of the reappearance of gap-junctional communication suggests that it may be involved in terminating proliferation.

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PubMed1988

Acetylcholine receptors and concanavalin A-binding sites on cultured Xenopus muscle cells: electrophoresis, diffusion, and aggregation.

Using digitally analyzed fluorescence videomicroscopy, we have examined the behavior of acetylcholine receptors and concanavalin A binding sites in response to externally applied electric fields. The distributions of these molecules on cultured Xenopus myoballs were used to test a simple model which assumes that electrophoresis and diffusion are the only important processes involved. The model describes the distribution of concanavalin A sites quite well over a fourfold range of electric field strengths; the results suggest an average diffusion constant of approximately 2.3 X 10(-9) cm2/s. At higher electric field strengths, the asymmetry seen is substantially less than that predicted by the model. Acetylcholine receptors subjected to electric fields show distributions substantially different from those predicted on the basis of simple electrophoresis and diffusion, and evidence a marked tendency to aggregate. Our results suggest that this aggregation is due to lateral migration of surface acetylcholine receptors, and is dependent on surface interactions, rather than the rearrangement of microfilaments or microtubules. The data are consistent with a diffusion-trap mechanism of receptor aggregation, and suggest that the event triggering receptor localization is a local increase in the concentration of acetylcholine receptors, or the electrophoretic concentration of some other molecular species. These observations suggest that, whatever mechanism(s) trigger initial clustering events in vivo, the accumulation of acetylcholine receptors can be substantially enhanced by passive, diffusion-mediated aggregation.

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