Biomedical subjects
Publications and source records attributed to M D Egger.
Recently obtained anatomical evidence supports the division of the rat cuneate nucleus (CN) into three rostrocaudal regions, with the middle region receiving a disproportionately greater share of the primary sensory input. The CN in the rat conforms to the basic rostrocaudal CN pattern described in other mammals, including cat, monkey and raccoon.
A simple growth algorithm is presented that deals with one feature of dendritic growth, the distance between branches. The fundamental assumption of our growth algorithm is that the lengths of dendritic segments are determined by the branching characteristics of the growing neurite. Realistic-appearing dendritic trees are produced by computer simulations in which it is assumed that: (1) growth of individual neurons occurs only at the tips of each growing neurite; (2) the growing neurite can either branch (as a bifurcation) or continue to elongate; (3) events at any one growing tip do not affect the events at any other growing tip; and (4) the probability of branching is a function only of the distance grown either from the cell body (if branching has not occurred) or from the previous branch point. An analytic solution of a differential equation based on these same assumptions produces a distribution of dendritic segment lengths that accurately fits an experimentally determined distribution of dendritic segment lengths of reconstructed neurons, accounting for about 89% of the sample variance. Our analysis indicates that, immediately following branching, the temporary suppression of further branching during dendritic growth may be an important mechanism for regulating the distance between branches.
Primary afferent projections from cutaneous afferents in the forelimb and hindlimb digits to the dorsal horn (DH) were examined using 4 tracers: (1) 25% free horseradish peroxidase (HRP), (2) 2.5% wheat-germ agglutinin conjugated to horseradish peroxidase (WGA-HRP), (3) a mixture of 25% free HRP and 2.5% WGA-HRP (WGA-HRP/HRP) or (4) 0.1% HRP conjugated to cholera toxin (B-HRP). The tracer was injected intracutaneously into the digits. Three to 4 days later, the rats were perfused transcardially, transverse sections (60-microns thick) were cut and the HRP was reacted using the tetramethyl benzidine (TMB) method. The location of the label was reconstructed by camera lucida drawings. In rats which received an injection of HRP alone, no label was detected in the DH. Rats injected with WGA-HRP had projection patterns similar to those injected with WGA-HRP/HRP. Patterns of labelling with WGA-HRP differed markedly from those with B-HRP. WGA-HRP labelled cutaneous afferents projecting to Rexed's laminae I-III, with the densest label in lamina II; in contrast, B-HRP labelled cutaneous afferents projecting to laminae II-V, with the densest label in laminae III-IV. These results indicate that, for cutaneous primary afferents projecting to the DH, WGA-HRP and B-HRP labelled different subpopulations of fibers, with the B-HRP-labelled subpopulation biased toward afferents of larger diameter. Rostrocaudally, the extent of the densest fiber projections, whether labelled by WGA-HRP or by B-HRP, was essentially the same, but the extent of the less densely labelled projections was much greater with B-HRP than with WGA-HRP. Comparisons of the projection maps from each of the five digits, using either WGA-HRP or B-HRP, indicated that, as seen in transverse sections through the DH, there was extensive overlapping among the labelled cutaneous afferent fibers from adjacent, or even non-adjacent digits.
To analyze the patterns of cutaneous primary afferent fibers projecting to the dorsal column nuclei in the rat, horseradish peroxidase (HRP)-based tracers were injected intracutaneously into a number of discrete regions of the forelimbs and hindlimbs. Three-4 days following the HRP injections, the rats were perfused transcardially; 60 microns transverse sections were cut, and the HRP was reacted using the tetramethyl benzidine method. Comparisons were made of projections following injections with choleragenoid-conjugated horseradish peroxidase (B-HRP) or with wheat-germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). B-HRP and WGA-HRP produced similar patterns of labelling, but B-HRP produced greater intensity of labelling and slightly larger projection areas. In the cuneate nucleus (CN), HRP labelling of primary afferents from small, delimited regions, e.g., from a portion of the skin of a single digit, appeared to be precisely restricted in rostrocaudally oriented columns, with little or no overlap (in the mediolateral and dorsoventral plane) into adjacent regions. With respect to rostrocaudal organization, a region in the CN containing a dense population of cutaneous primary afferent fibers appeared to be similar to the middle, or cluster, region in cats and in raccoons and the pars rotunda in primates. Projection patterns were very consistent from rat to rat, but their somatotopic organization differed from that suggested by electrophysiological studies: cutaneous afferents from forelimb digit 1 projected near the ventral border of the CN; those from digit 5 projected dorsomedially to those from digit 1; the projections from the remaining digits formed a crescent between the projections from digits 1 and 5. In the gracile nucleus, the organization of cutaneous afferent projections from hindlimb digits was more variable and complex than that found in the CN.
The tergotrochanteral muscles (TTM) in the second thoracic segment of the fruitfly, Drosophila melanogaster, power the jump-escape response. The cell bodies of the motoneurons innervating these muscles, located in the thoracico-abdominal ganglion, have prominent posterior and medial neurites. While in wildtype flies of the Canton-S (C-S) and Oregon-R (O-R) strains, the medial neurite of a TTM motoneuron rarely crossed the midline (C-S: 0/17; O-R: 2/8), in heterozygous flies with a deficiency at the base of the X-chromosome, Df(1)16-3-22, the medial neurite frequently crossed the midline (Df(1)16-3-22/C-S: 7/12; Df(1)16-3-22/O-R: 18/22).
A novel somatotopic map of primary cutaneous afferents projecting to the cuneate nucleus in the rat was determined by transganglionic transport of wheat-germ agglutinin conjugated to horseradish peroxidase and free horseradish peroxidase. Intracutaneous injections of tracer into different limited regions of the forelimb resulted in discrete areas of label for each injection site, with little or no overlap into other projection areas. The map of cutaneous projections onto the cuneate nucleus revealed by our anatomical tracing provided much more detail than any previous study in the rat, and demonstrated some significant differences from earlier maps based on electrophysiological recordings.
The tergotrochanteral (TTM) "jump" muscles in the second (T2) and third (T3) thoracic segments of the fruit fly, Drosophila melanogaster, were analyzed morphologically and morphometrically in wildtype (Canton-S) and bithorax mutants (abx bx3 pbx/Df(3R)P2). In the transformed T3 segments of mutant flies, the TTMs were greatly increased in fiber number (330% of wildtype), length (141%), and volume (460%), thus manifesting both hyperplasia and hypertrophy. In contrast, TTMs in the "untransformed" T2 segments of mutant flies were both hypoplastic and hypotrophic, in that significant decreases in fiber number (93% of wildtype), length (90%), and volume (80%) were observed. Two relationships emerged from analysis of the morphometric data: 1) Although the fiber numbers and volumes of the transformed T3 TTMs in bithorax flies were greatly increased, the total combined volumes of the TTMs in T2 + T3 remained approximately the same in bithorax compared to wildtype flies. 2) The changes in TTM volumes in bithorax flies compared to those in wildtype were proportional to the relative changes in fiber numbers times the relative changes in muscle lengths. These observations suggest that the genes of the bithorax complex influence the number and the length of tubular muscles fibers of the TTMs, but do not significantly affect the mean cross-sectional areas of these fibers. Fibrillar muscle fibers, which are not found at all in T3 segments in wildtype flies, were observed in the transformed T3 segments of bithorax mutants in 11 of 18 cases (61%), but typically as wisps, not in complete muscles. We suggest that, in the T3 segment of the bithorax flies, the relative differences between the massive transformation of tubular TTMs vs. the minimal appearance of fibrillar muscles may be related, in part, to the relative availability of muscle precursors.
We report on computer-assisted three-dimensional reconstruction of spinal cord activity associated with stimulation of the plantar cushion (PC) as revealed by [14C]-2-deoxy-D-glucose (2-DG) serial autoradiographs. Moderate PC stimulation in cats elicits a reflex phasic plantar flexion of the toes. Four cats were chronically spinalized at about T6 under barbiturate anesthesia. Four to 11 days later, the cats were injected (i.v.) with 2-DG (100 microCi/kg) and the PC was electrically stimulated with needle electrodes at 2-5 times threshold for eliciting a reflex. Following stimulation, the spinal cord was processed for autoradiography. Subsequently, autoradiographs, representing approximately 8-18 mm from spinal segments L6-S1, were digitized for computer analysis and 3-D reconstruction. Several strategies of analysis were employed: 1) Three-dimensional volume images were color-coded to represent different levels of functional activity. 2) On the reconstructed volumes, "virtual" sections were made in the horizontal, sagittal, and transverse planes to view regions of 2-DG activity. 3) In addition, we were able to sample different regions within the grey and white matter semi-quantitatively (i.e., pixel intensity) from section to section to reveal differences between ipsi- and contralateral activity, as well as possible variation between sections. These analyses revealed 2-DG activity associated with moderate PC stimulation, not only in the ipsilateral dorsal horn as we had previously demonstrated, but also in both the ipsilateral and contralateral ventral horns, as well as in the intermediate grey matter. The use of novel computer analysis techniques--combined with an unanesthetized preparation--enabled us to demonstrate that the increased metabolic activity in the lumbosacral spinal cord associated with PC stimulation was much more extensive than had heretofore been observed.
In the rat, the numbers and locations of motoneurons innervating the short plantar muscles of the hindlimb (supplied by the medial and lateral plantar nerves, as well as a branch of the sural nerve) were determined by using both horseradish peroxidase (HRP) and fluorochromes as retrograde labels. Topographical organization within the plantar motor nucleus was examined by exposing individually the cut ends (encapsulated in low melting-point paraffin) of medial plantar, lateral plantar, and sural nerves to HRP. In addition, double-labeling experiments were conducted in which the medial plantar nerve was labeled with one fluorochrome (either true blue or diamidino yellow) and the lateral plantar nerve with another. The plantar motor pool is located in the extreme dorsolateral portion of the ventral horn, usually concentrated in the fifth lumbar (L5) spinal segment. Labeled motoneurons extended caudally into the sixth lumbar (L6) segment and rostrally into portions of the fourth lumber (L4) segment. Motoneurons of the medial plantar, lateral plantar, and sural nerve have overlapping territories. Sural motoneurons (about 70 cells per side) are generally confined to L5, medial plantar motoneurons (about 180 cells per side) tend to be concentrated in caudal L5 and rostral L6, whereas the lateral plantar motoneurons (about 310 cells per side) extend throughout the entire length of the plantar motor pool. The distribution of motoneuronal cell size is unimodal (mean cross-sectional area = 610 +/- 150 microns2). Cell bodies of plantar motoneurons tend to have similar geometries in all three major planes of sectioning. In all, the combined plantar plus sural nerve population amounts to about 560 motoneurons on each side of the spinal cord. On the basis of these data, and those published by others, the innervation of the small muscles of the foot accounts for about 25% of the motor axons carried by the entire sciatic nerve.
Dorsal horn cells (DHC) in the spinal cord of cats responding to low threshold mechanical stimulation of the plantar cushion (PC), labelled with intracellular horseradish peroxidase, displayed a wide variety of anatomical and physiological characteristics, some of which were quite consistent within a given lamina. These DHC had cell bodies located medially in spinal segment L7, in laminae III, IV and VI. Dendritic fields varied greatly, for the most part conforming to patterns corresponding to their locations in the dorsal horn. Axon collaterals of most DHC ramified near their cell bodies, chiefly projecting ventral ward within the same lamina as the cell body or into more ventral laminae. Cells with the shortest central delays had cell bodies in laminae III and IV. Response latencies of DHC to PC stimulation were only slightly greater than the latencies of primary afferent fibers. Lamina IV cells had mechanical thresholds comparable to those of the primary afferents, significantly lower than the thresholds of cells of laminae III or VI. The receptive fields (RF) of the lamina IV cells were significantly larger than those of the primary afferent fibers or of the lamina VI cells. Relationships among RF area, mediolateral locations of cell bodies and dendritic spread were investigated.
Rhythmical whisking of the mystacial vibrissae at about 7 Hz during exploration is one of the most conspicuous behavioral patterns in the rat. To identify the final common pathway for vibrissal movement, individual motor branches of the facial nerve, including the posterior auricular, temporal, zygomatic, buccal, marginal mandibular, cervical, stylohyoid, and posterior digastric branches, were cut, either singly or in various combinations. We found that vibrissal movement could be abolished only by transection involving the buccal branch and the upper division of the marginal mandibular branch. To trace back the central origins of the buccal and marginal mandibular, as well as the other branches of the facial nerve, all distal to the stylomastoid foramen, horseradish peroxidase (HRP) was applied to the cut proximal ends of these individual branches. The retrograde HRP labelling in the facial motor nucleus revealed topographical representation of these branches in which the buccal and marginal mandibular branches were represented laterally. The stylohyoid and posterior digastric branches originated from cells in the suprafacial nucleus. Consistent with earlier observations with intramuscular HRP injections, the motoneuronal population devoted to vibrissal movement did not seem to be substantially larger than that for other facial movements. An additional examination was made of the labelled afferent component of the facial motor nerve. We confirmed and extended previous findings that none of the above facial motor nerve branches, except the posterior auricular branch, contained a significant number of afferent fibers originating from the geniculate ganglion, the sensory ganglion of the seventh nerve. In addition, no labelling was seen in the mesencephalic trigeminal nucleus or trigeminal ganglion. These findings, in combination, suggest that, with the exception of the posterior auricular branch, all the facial motor nerve branches, including those involved in vibrissal movement, are almost entirely efferent.
Systemic administration of 4-aminopyridine (4-AP) increased the size of the cutaneous receptive fields of 9 of the 15 dorsal horn cells tested. These receptive fields were on the feet and toes of the hind limbs of cats. Receptive field sizes increased with increasing doses of 4-AP. However, 4-AP administration did not change the responses of dorsal horn cells to graded mechanical stimuli administered near the centers of their receptive fields.
The intra-axonal horseradish peroxidase technique was used to examine the central terminals of 7 A beta primary afferent fibers from rapidly adapting (RA) mechanoreceptors in the glabrous skin of the cat's hindpaw. At the light microscopic level, labelled collaterals were seen to bear occasional boutonlike swellings, mostly (75-82%) of the en passant type. These swellings were distributed more or less uniformly from lamina III to a dorsal part of lamina VI in the dorsal horn, over a maximum longitudinal extent of about 4 mm. At the electron microscopic level, we observed that labelled boutons of RA afferent fibers were 1.0 to 3.3 micrometers in longest sectional dimension, and contained clear, round synaptic vesicles. They frequently formed asymmetric axospinous and axodendritic synapses and commonly appeared to receive contacts from unlabelled structures containing flattened or pleomorphic vesicles plus occasional large dense-cored vesicles. The examination of synaptic connectivity over the entire surface of individual boutons indicated that RA afferent boutons each made contacts with an average of one spine and one dendrite and, in addition, appeared to be postsynaptic to an average of two unlabelled vesicle-containing structures. This synaptic organization was, in general, more complex than that we had seen previously in Pacinian corpuscle (PC) and slowly adapting (SA) type I mechanoreceptive afferent fibers. Our findings indicate that RA, SA, and PC afferent terminals, while displaying some differential synaptic organizations, have many morphological and synaptological characteristics in common. These afferent terminals, in turn, seem to be generally distinguishable from the terminals of muscle spindle Ia afferents or unmyelinated primary afferents.
Using a method to visualize HRP-containing cells in the geniculate ganglion (GG) in situ after decalcifying surrounding bone, we found that about 30% of the total (about 1000) GG cells contributed sensory fibers to the posterior auricular branch of the facial motor nerve. These cells are relatively large for GG cells in general. The remaining facial motor nerve branches, including those involved in vibrissal movement, contained few sensory afferent fibers originating from GG cells.
Photographic, electrophysiological and neurobehavioral analyses were used to examine the contribution of trigeminal inputs to the behavioral organization of eating in the rat. During eating, jaw opening was always preceded by a period of perioral contact with the food source. Mechanical or electrical stimulation of oral and perioral areas in anesthetized animals elicited compound action potentials in the mylohyoid nerve (jaw-opener innervation) at short latencies and low stimulus intensities. Trigeminal orosensory deafferentation (sparing jaw muscle afferents and efferents) abolished or significantly reduced mouth opening during eating. We conclude that trigeminal orosensory inputs provide an essential link in the stimulus-response chain mediating eating in the rat.
The glabrous skin of the hindlimb of the cat contains 3 types of low-threshold mechanoreceptors: Pacinian corpuscles (PC), and slowly and rapidly adapting receptors. In the present study, 12 primary afferent fibers transmitting impulses from PC were injected intra-axonally with horseradish peroxidase (HRP) in the spinal cord to examine the morphology of their terminals in the dorsal horn. At the light microscopic level, terminal arborizations were observed in laminae II-VI of the dorsal horn, extending up to 7 mm rostrocaudally in and near the seventh lumbar segment. Bouton-like swellings, predominantly (67%) of the en passant type, were distributed in two discrete clusters, one concentrated rostrally in Rexed's laminae III-IV, and the other concentrated caudally in lamina V. At the electron microscopic level, a combination of morphometric and serial reconstructive analyses with 3 fibers revealed the following. Boutons labelled with HRP invariably contained clear round vesicles, approximately 40 nm in diameter. Labelled bouton sections had longest dimensions of 1.84 +/- 0.63 micron. Their shapes varied from rounded to elongated forms with occasional scalloped appearances. A majority (73%) of the contacts associated with HRP-filled boutons were made with dendritic spines and shafts. Thick postsynaptic densities were usually associated with these synapses, although thinner densities were also observed. 24% of the contacts made by labelled boutons were synapse-like contacts with unlabelled vesicle-containing structures. The vesicles in the unlabelled structures were usually pleomorphic, but sometimes round. These contacts were identified as 'synapse-like' because labelling obscured possible landmarks necessary for definitive identification of synapses. However, in most of these contacts, there was an accumulation of vesicles near the cleft on the unlabelled side, suggesting that the labelled boutons were postsynaptic. Only 3% of the contacts made by labelled boutons were axosomatic. The lengths of contacts with dendritic spines (0.49 +/- 0.23 micron) or with dendrites proper (0.45 +/- 0.20 micron) were significantly longer than those with vesicle-containing unlabelled structures (0.31 +/- 0.18 micron). The portions of cross-sectional bouton contours devoted to synaptic or synapse-like contacts accounted for 9-13% of the perimeters. The larger the bouton, the longer the summed lengths of contacts. Serial reconstruction of selected labelled boutons revealed both simple and quite complex synaptic organizations, including glomeruli with labelled boutons as the central component.(ABSTRACT TRUNCATED AT 400 WORDS)
Primary afferent fibers transmitting impulses from slowly adapting (SA) Type I receptors in the glabrous skin of the hind paw of the cat were injected intraaxonally in the spinal cord with horseradish peroxidase (HRP). At the light microscopic level, terminal arborizations were observed in the medial dorsal horn extending up to 6 mm rostrocaudally in and near the seventh lumbar segment. Boutonlike swellings labelled with HRP were distributed in clusters in Rexed's laminae III-VI. There was a tendency for the most dorsal clusters from an individual fiber to be located rostrally and for the most ventral clusters to be located caudally. At the electron microscopic level, a combination of morphometric analysis and serial reconstruction revealed the following: (1) All the boutons labelled with HRP contained predominantly clear, round synaptic vesicles, 40-50 nm in diameter. (2) Labelled boutons (n = 75) had cross-sectional longest dimensions of 1.72 +/- 0.53 micron (Mean +/- S.D.), perimeters of 4.95 +/- 1.52 micron, and areas of 1.18 +/- 0.59 micron 2. Their shapes in section varied from rounded to elongated forms. (3) The sizes of labelled boutons decreased significantly and linearly with depth from lamina IV to VI. The shapes of the bouton cross sections also became rounder with depth in the dorsal horn. (4) About 72% of synaptic contacts associated with HRP-filled boutons were with dendritic spines and shafts; most of these synapses were of the asymmetric type. (5) The remainder (28%) of the appositions were synapselike contacts between labelled boutons and unlabelled structures containing flattened or pleomorphic vesicles, and occasional dense-cored vesicles. (6) We observed no unequivocal axosomatic contacts made by labelled boutons. (7) The lengths of synaptic appositions with dendritic spines (0.46 +/- 0.20 micron) or with dendritic shafts (0.51 +/- 0.18 micron) were significantly greater than the synapselike contacts with vesicle-containing unlabelled structures (0.29 +/- 0.09 micron). (8) Complex neuropilar organization was occasionally seen with labelled boutons as central elements, although simpler organizations were much more common. In summary, HRP-labelled fibers ended predominantly in boutons containing clear, round vesicles forming axospinous and axodendritic synapses.(ABSTRACT TRUNCATED AT 400 WORDS)
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