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L T Landmesser

Publications and source records attributed to L T Landmesser.

30 records · Page 2Linked to original sources

Interspecies selective motoneuron projection patterns in chick-quail chimeras.

During normal development chick motoneurons have been shown to project selectively to appropriate muscles by responding to a series of cues, both specific and nonspecific, within the limb. We tested the ability of motoneurons from another avian species, the Japanese quail, to respond to these cues by transplanting chick limb buds onto quail embryos and quail limb buds onto chick embryos between stages 17 1/2 and 19. Feulgen staining, which distinguishes chick from quail cells on the basis of nuclear chromatin, revealed that all limb tissue, including muscle, was of donor origin, indicating that the migration of somite-derived muscle precursor cells had been completed by the time of transplantation. Normal quail motoneuron pools for most muscles were located in the same relative positions as homologous chick pools. In chick-quail chimeras we found that the motoneuron pools of one species selectively innervated the homologous muscles in the limb of opposite species with considerable precision. This was determined by defining the segmental innervation pattern of the muscles electrophysiologically and by retrogradely labeling motoneuron pools with HRP. Selective innervation was confirmed by using the functional activation patterns of the motoneuron pools as an additional means of identifying motoneurons. We conclude that any limb-derived cues required by motoneurons to project to their appropriate muscles must be similar in chick and quail and that the growth cones of both species must have similar detector systems for responding to these cues. Only 7 spinal segments were found to innervate the quail limb (versus 8 for the chick), resulting in an anterior shift in the spinal segments innervating several posterior quail muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

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Cell death of lumbosacral motoneurons in chick, quail, and chick-quail chimera embryos: a test of the quantitative matching hypothesis of neuronal cell death.

The quantitative matching hypothesis of neuronal cell death was tested for the chick hindlimb by determining the relationship between myotube number at the onset of motoneuron cell death and the number of motoneurons that survive in chicks, quail, and chick-quail chimeras. Hindlimb buds, which differ in size between the 2 species, were exchanged at stages 16 1/2-19, myosin ATPase-stained myotubes in selected thigh muscles were counted during the cell death period (stages 30-34), and lumbosacral motoneurons were counted following the cell death period (stage 38). No quail motoneurons were rescued when quail cords innervated chick limbs. When chick cords innervated quail limbs, the number of surviving motoneurons was significantly decreased but not to quail values. We consider that this occurred because chicks develop more slowly than quail, and we found that transplanted chick limbs were developmentally younger than the contralateral quail limb at the onset of motoneuron cell death and contained fewer myotubes. Similarly, transplanted quail limbs contained more myotubes at the onset of cell death than normal stage 30 quail limbs. An excellent correlation was obtained during normal development of both species between the number of myotube clusters at the onset of cell death and the number of surviving motoneurons. This correlation was also observed for chick-quail chimeras, and when the data points were plotted for control chick, control quail, chick host-quail limb, and quail host-chick limb, the correlation coefficient was 0.996. This strongly suggests that some parameter closely related to myotube number limits the number of motoneurons that will survive. A proposal consistent with our observations is that motoneuron survival is dependent on the uptake of a myotube-derived trophic factor that can only be taken up at synaptic sites and that the number of such sites is limited and directly related to myotube number. In conclusion, our observations strongly support a quantitative-matching component in the process of neuronal cell death. However, since we were unable to rescue any neurons, we cannot exclude the possibility that some proportion of neurons normally dies for reasons other than peripheral competition.

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Development of the major pathways for neurite outgrowth in the chick hindlimb.

To elucidate mechanisms that may control development of the gross anatomical nerve pattern, motoneuron outgrowth into the chick hindlimb was examined using orthograde labeling, scanning and transmission electron microscopy, and Alcian blue staining. Results show that growth cones are not guided by contact with oriented extracellular fibrils, aligned mesenchyme cells, the myotome, or the vasculature. Pathways are not delineated by cell-free space or channels of lower cell density; however, densely packed mesenchyme may form barriers that channel outgrowth. In addition, abundant mesenchymal cell death was seen at the nerve front. This cell death may provide space that encourages growth cone advancement. Pathways often lie along interfaces between areas that stain darkly and lightly with Alcian blue, which specifically stains glycosaminoglycans, and growth cones never penetrate areas that stain intensely, such as the pelvic girdle, which is known to be a barrier to outgrowth. Leading growth cones form specialized contacts with mesenchyme cells, but the predominant contacts are interneuronal. It is proposed that the anatomical pattern of outgrowth is determined by the distribution of preferred substrata, the most preferred substratum being other neurites. Further, neurites tend to prefer loose mesenchyme to dense mesenchyme or areas rich in glycosaminoglycans.

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Specificity of early motoneuron growth cone outgrowth in the chick embryo.

During development, chick lumbosacral motoneurons have been reported to form precise topographic projections within the limb from the time of initial outgrowth. This observation implies, first, that motoneurons select the appropriate muscle nerve pathway and, second, that they restrict their ramification within the primary uncleaved muscle masses to appropriate regions. Several reports based on electrophysiology and orthograde horseradish peroxidase (HRP) labeling have shown muscle nerve pathway selection to be fairly precise. However, studies based on retrograde labeling with HRP have produced conflicting reports on the extent to which vertebrate motoneurons make projection errors. Since it is difficult to distinguish between true projection errors and HRP leakage when using retrograde labeling, we decided to assess the distribution of labeled growth cones in 25-micron serial plastic sections, following orthograde labeling of identifiable subpopulations of motoneurons during the period of initial axon outgrowth. Examination of a large number of muscle nerves revealed no segmentally inappropriate axons, confirming earlier reports that muscle nerve pathway selection is very accurate. In addition, we observed that growth cones take widely divergent trajectories into the same muscle nerve, suggesting that growth cones are responding independently to some specific environmental cue rather than being passively channeled at this point. The distribution of labeled growth cones within the muscle masses provided direct evidence that motoneurons did not at any time project to obviously inappropriate muscle regions.(ABSTRACT TRUNCATED AT 250 WORDS)

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Growth cone morphology and trajectory in the lumbosacral region of the chick embryo.

We quantitatively analyzed several features of orthogradely labeled peripheral growth cones in the lumbosacral region of the chick embryo. We compared motoneuron growth cones in regions where they appear to express specific directional preferences (the plexus region and regions where muscle nerves diverge from main nerve trunks), which we operationally defined as "decision regions," to motoneuron growth cones in other pathway regions (the spinal nerve, nerve trunk, and muscle nerve pathways) which we termed, for contrast, "non-decision region." We found that motoneuron growth cones are larger, more lamellepodial, and have more complex trajectories in decision regions. Sensory growth cone populations, which are thought to be dependent upon motoneurons for outgrowth (Landmesser, L., and M. Honig (1982) Soc. Neurosci. Abstr. 8: 929), do not enlarge or become more lamellepodial in motoneuron decision regions, suggesting that this local environment does not affect all species of growth cones equally and that the alterations in motoneuron growth cones in these regions may be relevant to their specific guidance. In addition, the resemblance between the sensory population and other closely fasciculating growth cones lends support to the suggestion that sensory neurons utilize motoneuron neurites as a substratum. We suggest that the convoluted trajectories, enlarged size, and more lamellepodial morphology of motoneuron growth cones in decision regions is either related directly to the presence of specific cues that guide motoneurons or to some aspect of this environment that allows them to respond to specific cues.

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Activation patterns of embryonic chick hind limb muscles recorded in ovo and in an isolated spinal cord preparation.

Muscle activation patterns of embryonic chick hind limb muscles were determined from electromyographic (e.m.g.) recordings in an isolated spinal cord/hind limb preparation of stage 34-36 embryos, and were compared with in ovo e.m.g. activity from similarly staged embryos. Muscle activity in ovo consisted of periodically recurring sequences of bursts during which antagonistic muscles often alternated and synergistic muscles were co-active, as compatible with their mature function. However, more variable behaviour was also observed. Burst sequences in ovo were often initiated by a short-duration, high-amplitude discharge that occurred synchronously in all muscles studied, and which was followed by a period of electrical silence that was longest in the flexor muscles. This type of activity has not been described previously in mature animals. In ovo movement sequences were generally initiated by extensor activity which progressively declined in duration and intensity throughout the sequence, while flexor activity progressively intensified. The onset of activity in extensor muscles was accompanied by an abrupt decrease in flexor activity, whereas the converse was not observed. Spontaneous movement sequences also occurred when the spinal cord and hind limb were isolated and maintained in oxygenated Tyrode solution for several hours. Deafferentation experiments indicated that the motor pattern in this preparation was generated centrally by circuits within the spinal cord. Activity from the isolated cord was less variable than that occurring in ovo, consisting of sequences of highly regular recurring bursts. Each burst began with a brief high-amplitude discharge that occurred synchronously in all muscles and which was similar to that observed in ovo. This was followed by a silent period, which was longest in the flexors, and then by a more prolonged burst. Although its behaviour differs from that in ovo in some respects, it is concluded that the isolated cord maintained in vitro produces a spontaneous and patterned motor output.

Action Potentials↗

The activation patterns of embryonic chick motoneurones projecting to inappropriate muscles.

Chick lumbosacral motoneurones were caused to innervate foreign muscles by surgically rotating or shifting the limb bud about the anterior-posterior axis in stage 17-18 embryos. The activation pattern of such wrongly projecting motoneurones was assessed at stages 35-38 by recording electromyographic activity from muscles in an isolated spinal cord/hind limb preparation. Muscle activity was classed as flexor- or extensor-like according to the characteristics of the patterned sequence of bursts elicited by a single shock to the thoracic cord. Wrongly projecting motoneurones did not have their activation pattern altered to one appropriate for the muscle innervated; therefore in some cases a particular muscle was activated with a pattern similar to its original one, and in other cases in an opposite manner. Mixed flexor-extensor-like activation of a single muscle was, however, rare. The identity of motoneurones projecting to a muscle was determined by their cord location following retrograde labelling with horseradish peroxidase. This allowed us to conclude that motoneurones could develop their normal pattern of activation even when projecting to foreign muscles. It is concluded that the cord circuits (presumably composed of local interneurones responsible for the activation of motoneurones in the isolated cord preparation are not altered by retrograde influences from the muscle. Wrongly projecting motoneurones, which were maintained throughout the normal cell death period, were activated during spontaneous embryonic movements, and in many cases were found to have a behaviourally inappropriate activation pattern. These observations are discussed in relation to proposed mechanisms by which developmental errors in connectivity are corrected.

Action Potentials↗

Pattern and specificity of axonal outgrowth following varying degrees of chick limb bud ablation.

Motoneurons grow into the chick hindlimb via consistent pathways, within which they make specific decisions leading to their correct targets. To determine which axonal guidance features dictate the position of the pathways and to examine the distribution of specific cues, we totally or partially ablated the early hindlimb bud and determined how the subsequent pattern of nerve outgrowth related to the distribution of tissue remnants. Our results suggest that local elements determine the gross anatomical pattern of outgrowth. First, determinants of individual pathways could be selectively removed without altering the pattern in other regions. Second, neurites were restricted to the plexus region at the base of the leg (within which, for unknown reasons, they proceeded posteriorly) unless distal permissive pathways or nearby target remnants were present. Finally, we found that the central region of the pelvic girdle, adjacent to the plexus region, determines the position where the major nerve trunks enter the leg. When gaps were introduced in this region of the girdle, nerves traversed the gaps and directly innervated adjacent muscle. The developing girdle is probably a nonpermissive environment for axon elongation, and axons enter the leg only where it is locally absent. Our results also support the concept that the specific cues that neurites use to reach their appropriate muscles are local. We find that neurites could make correct and specific decisions in the plexus region in the absence of all tissues distal to the pelvic girdle. This shows that the cues for these decisions are independent of the target and must reside in the local mesenchyme. In addition, when muscle remnants were present they were correctly innervated.(ABSTRACT TRUNCATED AT 250 WORDS)

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Synaptic plasticity: keeping synapses under control.

Recent studies have confirmed that a retrograde signal is produced at the neuromuscular junction that can adjust the efficacy of transmission to meet long-term changing needs. Genetic manipulations in Drosophila have begun to define the circumstances in which such signals are generated and how they act.

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