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M B Heaton

Publications and source records attributed to M B Heaton.

At least 73 records · Page 4Linked to original sources

The influence of muscle-conditioned media on chick embryo brainstem neurons in culture.

Brainstem pieces from the trigeminal region of the metencephalic basal plate of 10-day chick embryos were dissociated and cultured in control conditions or in the presence of muscle-conditioned medium (MCM). The MCM was derived from age-matched target tissue relevant to this neuronal region (jaw musculature), from relevant target tissue of an age at which innervation would initially be taking place (4 days), and from nonrelevant target tissue also of an early stage (4-day limb bud). Neuronal survival and differentiation was assessed daily, for 7 days. Survival and differentiation were significantly enhanced by the 4-day jaw MCM compared to both the controls and the cultures grown with 10-day jaw MCM and 4-day limb MCM. These measures in the presence of 10-day jaw MCM and 4-day limb MCM did not differ, but surpassed that seen in control cultures. The results are compared to the more specific responsiveness seen in earlier (2-day) neural tube cultures, and their relationship to in vivo regenerative nerve fiber outgrowth is considered.

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Specific responsiveness of chick trigeminal motor nucleus explants to target-conditioned media.

Explants of the neural tube from stage 11 chick embryos containing the metencephalic trigeminal (V) motor nucleus were cultured in standard control medium, in medium conditioned by appropriate target musculature (the mandibular process of the first visceral arch that gives rise to jaw musculature innervated by motor V) or in medium conditioned by inappropriate target musculature (rostral limb bud tissue). The appropriate and inappropriate muscle tissues were of the same developmental stage (stage 22) and were in similar states of differentiation. At this point in vivo, both are just beginning to be innervated. The neuritic outgrowth from the explants was quantified after 6 days in vitro. While explants from all three groups appeared healthy and exhibited some neuritic outgrowth, the density and complexity of this growth was significantly greater in the group cultured with the appropriate (jaw) muscle-conditioned medium. Growth in this group significantly surpassed that of both the control and the inappropriate muscle-conditioned medium group did not differ from the control group. These results demonstrate a specific responsiveness of the trigeminal motor nucleus population to its appropriate target tissue. Since relatively small amounts of the muscle-conditioned medium were used with each explant, it is suggested that there is a high degree of sensitivity of this population to factors present in their target at the time innervation would normally be occurring. It is hypothesized that such selective responsiveness may play a role in guiding or sustaining growth during normal neurogenesis.

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The influence of target tissue age on neurite outgrowth from chick embryo trigeminal motor nucleus explants.

In a previous study, explants from the neural tube of 2-day chick embryos, containing the trigeminal motor nucleus, showed specific enhancement of neurite outgrowth in the presence of culture medium conditioned by target musculature of an age at which innervation would normally be taking place in vivo. In the present study, similar explants were grown in the presence of target-conditioned medium from midincubation (10-day) embryos and late-incubation (18-day) embryos. At both of these later developmental stages, the target muscle-conditioned medium failed to produce enhanced neurite outgrowth. Rather, fewer processes were extended from the trigeminal explants in both experimental conditions compared to control cultures. This inhibitory effect was statistically significant in some cases, and did not appear to be related to the amount of target-conditioned medium present. It is concluded that trophic influences on nerve fiber outgrowth by target tissue are stage-specific in this system, and it is speculated that inhibitory influences such as those demonstrated may interact with excitatory influences in the normal patterning of the developing nervous system.

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Patterns of extraocular innervation by the oculomotor complex in the chick.

The horseradish peroxidase retrograde tracer technique was used to map the projection pattern of the oculomotor nuclear complex to the extraocular muscles in the chick embryo. The following projection pattern was found: The dorsolateral oculomotor subnucleus innervates the ipsilateral inferior rectus muscle, the dorsomedial subnucleus innervates the ipsilateral medial rectus muscle, a lateral division of the ventromedial subnucleus innervates the ipsilateral inferior oblique muscle, and a medial division of the ventromedial subnucleus innervates the contralateral superior rectus muscle. The so-called central nucleus also innervates the contralateral superior rectus muscle. This pattern was extremely discrete, with virtually no overlapping representations. These results provide the first evidence for a functional medial-lateral subdivision of the ventromedial subnucleus. This pattern relates to the unusual development of this subnucleus and suggests that only part of the primordium for this cell group migrates across the midline during its ontogeny, rather than all of it, as was previously believed. The subnuclear organization of the avian oculomotor complex is also considered in comparison to such functional organization in other species.

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Developmental relationships between trigeminal ganglia and trigeminal motoneurons in chick embryos. I. Ganglion development is necessary for motoneuron migration.

The migration and early development of trigeminal (V) motoneurons were studied in chick embryos in which two different populations of primary trigeminal sensory neurons had been removed prior to the birthdate of the V motoneurons. Ablation of mesencephalic neural crest cells, which eliminates monosynaptic sensory input, did not affect the migration, early development, or later differentiation of the V motoneurons. However, when the anlagen of the V ganglion were removed, the V motor root did not exit from the brainstem and the V motor nucleus did not develop. Although the neurons of the V ganglion do not innervate adult V motoneurons, these populations are related developmentally. In those embryos in which the V ganglion did not develop, medial column cells, which are midline, postmitotic, premigratory V motoneurons, and a few medial, elongated cells (possibly migratory) were present until days 5-6, but these cells did not complete their lateral migration to form the lateral nucleus of V. In cases where the ganglion anlagen were not completely removed, the number of postmigratory V motoneurons was positively correlated to the size of the ganglion remnant. There also was a correlation between the axial position of the postmigratory V motoneurons and the ganglion remnants. If a caudal remnant developed, only caudal V motoneurons, whose axons reached the ganglion, migrated; if a rostral remnant developed, only rostral V motoneurons, with axons reaching this remnant, migrated. Additionally, if the central axons of the ganglion remnant entered the metencephalon in either dorsal or ventral ectopic positions, the V motor nucleus was located in a corresponding aberrant position. Thus, some characteristic of the V ganglion cells appears to guide the motor axons and somas to their final brainstem position.

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Developmental relationships between trigeminal ganglia and trigeminal motoneurons in chick embryos. II. Ganglion axon ingrowth guides motoneuron migration.

In the chick embryo the trigeminal (V) sensory ganglion cells send axons into the metencephalon a few hours before the V motoneurons migrate from the midline to form a lateral nucleus adjacent to the ingrowing sensory axons. This relationship suggests that the ganglion axons may influence the initiation and direction of V motoneuron migration. In the present experiment the development of the ganglion axons was retarded by removing the neural crest anlage of the V ganglion. Subsequently, V ganglion cells which were derived from the ectodermal placode anlage sent axons into the metencephalon up to 2 days later than normal. The lateral migration of the V motoneurons was similarly delayed, commencing only after the central axons from the placodal ganglia penetrated the metencephalon. This study demonstrates that the presence of V ganglion perikarya alone is not sufficient to guide the appropriate migration of V motoneurons. This migration occurs only after the axons from the V sensory ganglion cells have penetrated the brainstem.

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Developmental relationships between trigeminal ganglia and trigeminal motoneurons in chick embryos. III. Ganglion perikarya direct motor axon growth in the periphery.

The previous study in this series demonstrated that the ingrowth of the central axons of the trigeminal (V) ganglion is prerequisite to V motor axon outgrowth and somatic translocation. In the present experiment we determined whether further interactions with V ganglion cell bodies were required by V motoneurons after the V ganglion innervates the brainstem. Soon after the ganglion axons had penetrated the brainstem they were severed, and a barrier, either permeable or impermeable, was placed between the ganglion cell bodies and the metencephalon. V motor axons grew along aberrant pathways to circumvent the impermeable barriers, many rerouting to reach the V ganglion. Only those V motor nerves which contacted the V ganglion distal to the barrier reached their target musculature in the mandible. The pattern of migration of V motoneurons was normal regardless of the V motor nerve trajectory, but the cell bodies of those axons which did not reach a muscle were not fully differentiated. When permeable barriers (Millipore filters) were implanted, the nerves followed two types of trajectories. If the pore size of the filter was small (0.45 and 0.025 microns), the V motor nerves grew identically to those observed in embryos in which impermeable barriers had been implanted. If the pore size of the filter was large (8.0 and 0.08 microns), the V motor nerve grew along its normal path directly to the barrier. Small axonal bundles from these nerves frequently grew into the filter toward the distal V ganglion. These results indicate that V motor axons preferentially grow to the V ganglion perikarya after exiting from the brainstem. Contact with the V ganglion always results in V motor nerve growth to the mandible while growth of the V motor axons to aberrant target sites only occurs when the axons fail to contact the V ganglion cells distal to the barrier.

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A technique for improved resolution of [3H]thymidine autoradiography in the avian embryo: a preliminary report.

A technique is described for producing pulse-like effects in [3H]thymidine autoradiography in chick embryos. This procedure involves combining thymidine with reserpine, which temporarily inhibits thymidine incorporation. The concept is to introduce thymidine, allow time for its incorporation, then introduce reserpine, so that subsequent uptake is inhibited, and a relatively discrete label will appear in cells generated at the time of thymidine administration. The best results occurred when thymidine was administered 12 h prior to reserpine, or when the two were introduced simultaneously. A dose of 0.004 mg of reserpine produced the suppression, but had no deleterious effects, in terms of embryonic survival or in long-lasting changes in cell numbers, as reflected by cell counts in the trochlear nucleus, a population which was undergoing proliferation at the time of reserpine injection. Thus, this technique appears to hold considerable promise for improving the precision of the autoradiography procedure in avian embryos.

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The development of the oculomotor nuclear complex in the Japanese quail embryo.

The development of the oculomotor nuclear complex was studied in the Japanese quail. In hatchlings, this complex was found to consist of four subnuclei: The accessory, the dorsolateral, the dorsomedial, and the ventromedial. An arciform subnucleus or central subnucleus was not found in this species. All subnuclei were made up of homogeneous cell populations. The oculomotor primordia can first be recognized at day 3 of incubation, and a clear subdivision within this primordia is apparent on day 5 (total incubation period = 17 days). At this time the accessory subnucleus can be discerned. By day 6, a horizontally oriented dorsal cell mass, the anlagen of the dorsolateral and dorsomedial subnuclei, is seen, as is a vertically oriented ventral cell mass, the anlagen of the ventromedial subnucleus. The boundaries between the subnuclei are not yet distinct at this age, however. By day 7 of incubation, all four subnuclei can be detected the ventromedial subnuclei, are first seen on day 5, at which time the oculomotor commissure begins to appear. Migratory traffic is extensive during days 6 and 7 of incubation, with wide bands of migrating cells typically seen, spanning the entire dorsal-ventral extent of the oculomotor complex. These cells do not appear to be associated exclusively with the ventromedial subnuclei. In many instances, migratory cells appear to be affiliated with the dorsomedial cell group. By day 10 of incubation, migration has ceased and the oculomotor complex has attained its definitive configuration. These observations are discussed with comparative reference to earlier studies of chick and duck embryos and hatchings.

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Recovery from experimentally induced problem-solving deficits in neonatal Peking ducklings as a function of environmental stimulation.

Neonatal Peking ducklings were reared in 3 different environments containing varied light and sound stimulation. One group was exposed to the stimulative environment only during the last 3 prenatal days, 1 group was placed in the stimulative environment at the time of hatching, and the 3rd group was exposed to the stimulative environment both pre- and postnatally. No differences in performance in a detour learning task were noted between controls and experimentals from the 1st 2 groups (prenatal stimulation only and postnatal stimulation only); however, the group receiving both pre- and postnatal stimulation solved the detour task significantly faster than their controls. This learning improvement as a function of the stimulative environment represents a partial recovery from the deficit produced as a result of the experimental paradigm in which the shell covering the air space was replaced with clear plastic wrap, a procedure which has been shown to reduce oxygen consumption and lead to learning retardation.

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Operant escape learning in decerebrate duck embryos.

Peking duck embryos were trained in an operant escape task on Day 25 of incubation (total incubation period = 27 days) following decerebration on Day 24 of incubation. Experimental embryos successfully acquired the operant response, which involved the performance of a discrete foot flexion in order to terminate a shock applied to the wing. The decerebrate embryos resembled normal, unoperated embryos of this age in their acquisition of the operant behavior. This study demonstrates that this form of learning can be acquired without benefit of telencephalic structures, at least at this point in development. Differences in the consequences of decerebration during prenatal or neonatal versus adult stages are discussed.

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Early development and migration of the trigeminal motor nucleus in the chick embryo.

The development of the trigeminal motor nucleus in the chick embryo was studied using autoradiographic, cell staining, fiber staining, and axonal transport techniques. It was found that this nucleus arises very early in neurogenesis, with the first cells produced at 48 hours of incubation (stage 12), peak cell production at 50--56 hours (stage 15), and neuroblast proliferation completed by 72 hours (stage 18). As has been described in mammalian embryos, the primordial trigeminal cells move from the ventricular layer to accumulate as part of the common medial column, and later migrate in a ventrolateral direction to form the definitive lateral motor nucleus. The first identifiable component of the trigeminal system is the semilunar ganglion, which flanks the neural tube at stage 12, and sends afferents into the metencephalon by stage 13. By stage 12-13, the medial column cells are first apparent, and at stage 14, a few of these medial column cells have moved to begin formation of a lateral nucleus. At this time, a thin motor root can be seen exiting the brainstem. During subsequent stages, migratory traffic from medial to lateral regions increases, with cells frequently moving in association with fiber processes in the marginal zone. These fibers are presumed to emanate from secondary sensory, reticular, and medial column neuroblasts. By day 5, the medial column is greatly depleted and by day 6--7, the definitive lateral motor nucleus is formed. Beginning at 5 days, the dorsal motor nucleus can be detected, with cells from the lateral nucleus appearing to stream in a dorsomedial direction for its formation. Injections of horseradish peroxidase (HRP) into the mandibular process of the first visceral arch resulted in retrograde labeling of lateral nucleus cells as early as 3.5 days of incubation. In addition, migrating cells, intermediate between medial column and lateral nucleus, were similarly labeled. These observations indicate that processes of the lateral nucleus cells and those of migrating cells are well into their peripheral field at this age, but we cannot conclude that neuromuscular affiliations have been established, due to the possibility of HRP diffusion and growth cone uptake.

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Learning deficits in the neonatal Peking duckling produced by decreased oxygen consumption during late prenatal periods: a cautionary note.

Neonatal Peking ducklings were tested in a detour problem-solving task following replacement of the shell overlying the air space of the egg on Prenatal Day 24 (total incubation period: 27 days) with a plastic wrap. This procedure, a common practice in behavioral embryology, resulted in a significant decrease in oxygen consumption, and a protracted retardation in the mastery of the learning task, with experimental ducklings solving the problem at a mean age of 5.27 days, compared with 2.32 days for controls. These results serve to underscore the problems that may be encountered as a result of any interference with the integrity of the egg during the prenatal period.

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