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PubMed1991

Distribution and morphological characteristics of axons in the periodontal ligament of cat canine teeth and the changes observed after reinnervation.

The distribution and morphological characteristics of myelinated and non-myelinated axons innervating the lower canine periodontal ligament (PDL) in adult cats have been analysed. After perfusion fixation and decalcification, the teeth were slit transversely, divided into segments, and embedded in plastic. Ultrathin sections of each segment were examined in the electron microscope and used to reconstruct the whole PDL at 1, 4, 7, and 9 mm from the tooth apex. One millimeter from the tooth apex there were a mean of 920 myelinated axons and 1,415 non-myelinated axons. The numbers of axons declined toward the tooth crown. Bundles of myelinated and small non-myelinated axons lay adjacent to the blood vessels midway between the bone and cementum. Isolated myelinated axons appeared to have split away from these main nerve bundles and entered the avascular zone of the ligament, where they lost their myelin sheaths to become large non-myelinated axons rich in mitochondria. These non-myelinated axons sometimes appeared to be linked to collagen fibres and were thought to be the mechanoreceptor terminals. Twelve weeks after sectioning and inferior alveolar nerve, the total number of axons innervating the periodontal ligament was 50% of that found in the contralateral controls. The large non-myelinated axons had smaller mean diameters and contained fewer mitochondria, a change which may be consistent with a reduction in mechanoreceptor excitability.

Animals↗
PubMed2004

Three-dimensional computational reconstruction of lateral skull base with plastinated slices.

The goals of this study were to build the 3D reconstructed model of lateral skull base and to explore the spatial relationships of the important structures for providing the morphological basis for lateral skull base surgery and clinical image diagnosis. Blocks with edges of about 80 mm containing the lateral skull base region and adjacent structures were sawn out from both sides of the heads and sectioned on transverse plane at a thickness of 700 microm using a plastination technique. On an SGI workstation, a Contours-Marching cubes algorithm was selected to reconstruct the 3D model of the lateral skull base. Accurate alignment of the structures in the serial macroscopic sections was obtained by the employment of the plastination technique. The quality of the reconstructed images was distinct and perfect, specifically, the spatial positions and complicated adjacent relationships of various structures of the lateral skull base can be shown in direct viewing when they are displayed in background of the cranial bony substance. The time spent in displaying or rotating one image including 50 sections was 1.5 sec; all reconstructed structures can be represented individually or jointly and rotated in any plane. The plastination technique and computer-aided 3D reconstruction have an obvious advantage in the study of the complex anatomy of the lateral skull base. Plastination technique provides cross-section images of a higher resolution than those obtained from CT scanning. The computerized 3D reconstruction is important in studying the spatial anatomy of the lateral skull base and can serve as a standard for models created with other techniques.

Epoxy Resins↗
PubMed2002

Re-inventing anatomy: the impact of plastination on how we see the human body.

Over the past 20 years the development of plastination has opened up new vistas for gross anatomy. In particular, it has led to a major expansion in the range of human anatomic specimens available for teaching and its potential value in research is increasingly being appreciated. More recently, it has burst into the public arena through what has become known as 'Anatomy Art,' as depicted in the von Hagens exhibition, Körperwelten (Bodyworlds). In this exhibition, the lifeless cadavers of the dissecting room have been transformed into standing, sitting, and jumping lifelike plastinated 'models' that demonstrate spinal cords, tumorous lungs, cirrhotic livers, joint prostheses, and sagittally sectioned whole bodies. Not surprisingly, the exhibition has raised considerable ethical debate about treating human cadavers in this way, an issue of particular relevance to anatomists. This article is an attempt to further this debate by considering the nature of plastinated human specimens, and the context within which they should be examined. The only rationale for displaying (plastinated) human material in the public domain is an educational one, with a basis in a museum ethos. The boundaries of this educational rationale are discussed, as are the opportunities and challenges presented by plastination to the anatomical community.

Anatomy, Artistic↗
PubMed2002

Aortic arch variation analyzed by using plastination.

Different ramification patterns can be observed during the development of the aortic arch. In this study a common trunk (CT), which subsequently branched into the brachiocephalic trunk (BT) and left common carotid artery (LCCA), arose from the aortic arch. The LCCA arose from the CT 10.27 mm above the aortic arch. After crossing the ventral aspect of the trachea and esophagus, the LCCA became situated on the left side of the esophagus. The caliber and length of the main branches of the aortic arch were determined and compared to reports in the literature. This variation was discovered in the context of producing transverse body slices using an E12 plastination process.

Aged↗
PubMed2007

Three-dimensional reconstruction of the ankle by means of ultrathin slice plastination.

Computerized reconstruction of anatomical structures is becoming very useful for developing anatomical teaching modules and animations. Although databases exist consisting of serial sections derived from frozen cadaver material, plastination represents an alternate method for developing anatomical data useful for computerized reconstruction. Plastination is used as an excellent tool for studying different anatomical and clinical questions. The sheet plastination technique is unique because it offers the possibility to produce transparent slices series, which can easily be processed morphometrically. The purpose of this study was to describe a method for developing a computerized model of the human ankle using plastinated slices. This method could be applied to reconstruct any desired region of the human body.A human ankle was obtained, plastinated, sectioned, and subjected to 3D computerized reconstruction using WinSURF modeling system (SURFdriver Software). Qualitative observations revealed that the morphological features of the model were consistent with those displayed by typical cadaveric specimens. Morphometric analysis indicated that the model did not significantly differ from a sample of cadaveric specimens. These data support the use of plastinates for generating tissues sections useful for 3D computerized modeling.

Ankle↗
PubMed2007

Anatomical investigations and their ethical dilemmas.

The multi-faceted nature of modern anatomy comes as a surprise to many, especially when confronted by such seemingly different topics as cadavers and human embryo research. However, even these disparate facets of anatomy are linked by common underlying ethical considerations. This article traces historical views of anatomy and places them alongside the more contemporary dimensions of whole-body plastination, use of human material obtained under unethical circumstances, and human embryo research. These dimensions introduce issues of respect, human dignity, consent, scientific integrity, and societal expectations.

Anatomy↗
PubMed1981

Fine structure of myelinated mechanical nociceptor endings in cat hairy skin.

High-threshold mechanoreceptors (mechanical nociceptors) with myelinated axons were electrophysiologically identified in hairy skin of the cat as described by Burgess and Perl ('67). Such elements possess receptive fields consisting of a number of punctate areas from which maximal firing can be elicited by intense (skin-damaging) mechanical stimuli. The spots of the receptive field are separated from each other by unresponsive regions, i.e., by skin areas from which responses cannot be evoked by stimuli effective at the spots. Fine steel pins were inserted to bracket closely a number of the spotlike responsive areas for each of several units. After aldehyde perfusion of the animal, osmification of the tissue and embedding in plastic, the marked skin zones were examined in semithin and ultrathin sections at the light and electron microscopic level. Near each delineated area, a thinly myelinated axon was found that could be traced to the papillary layer where it loses its myelin sheath. Unmyelinated axons accompanied by thin Schwann cell processes were then traced and found to penetrate the epidermal basal lamina in one of the papillae. At the epidermal penetration site, the axons contained both clear round, and large, dense core vesicles; at this level, the surrounding Schwann cell cytoplasm exhibited numerous pinocytotic vesicles. The zone of penetration may constitute the receptive apparatus. Some of these axons have been traced within the basal epidermal layer where they become surrounded by keratinocytes, lose their Schwann sheath, and apparently terminate. This overall morphological pattern was consistently present in the demarked areas of focal responsiveness, and was rare in the surrounding skin; this and its difference from other cutaneous neural endings suggest that the intraepidermal axon-Schwann cel complex constitutes the receptive structure for myelinated mechanical nociceptors. It is suggested that such complexes are the sense organs responsible for initiating the sensation of pricking pain produced by localized mechanical injury of the skin.

Afferent Pathways↗
PubMed1985

Variability in the terminations of GABAergic chandelier cell axons on initial segments of pyramidal cell axons in the monkey sensory-motor cortex.

Chandelier cell axons were studied in the sensory-motor cortex of adult monkeys. The axonal fields of Golgi-impregnated chandelier cells in layer II in motor cortex are flattened sagittally. The vertical terminal portions of the axons varied both in length and in the numbers converging to form terminations of greater or lesser complexity. Golgi-impregnated plexuses were embedded in plastic and resectioned serially at 2.5-3.0 micrograms. A single axonal field could have as many as 400 terminal rows. All lie 3-13 micrograms beneath pyramidal cell somata. These terminations are not randomly distributed but instead, form clusters. Further resectioning the plastic sections for electron microscopy revealed that all the terminations are on the initial axon segments of pyramidal cells and all form symmetric synaptic contacts. In immunocytochemical material stained for glutamic acid decarboxylase (GAD), the enzyme involved in the synthesis of GABA, GAD-positive boutons were found to form symmetric synaptic contacts with a variety of postsynaptic elements including the axon hillocks and axon initial segments of pyramidal cells. Serial reconstructions from electron micrographs revealed GAD-positive terminals synapsing with the axon initial segment of pyramidal cells joined by cytoplasmic bridges and forming vertically oriented rows identical to those of chandelier cell terminals identified positively in the resectioned Golgi material. The GAD-positive terminals forming initial segment synapses were never continuous with GAD-positive terminals forming axo hillock synapses. The latter probably arise from basket cell axons. Initial segments of pyramidal cell axons in layers II and III were contacted by more GAD-positive terminals than the initial segments of pyramidal cell axons in layer V. The largest pyramidal cells in layer III received the most synapses. Many larger pyramidal cells, identified as callosally projecting cells by the retrograde transport of horseradish peroxidase (HRP), were shown in serial electron micrographs to possess large numbers of initial segment synapses, comparable to those seen in the immunocytochemical material. Serial reconstructions of pyramidal cell axons from axon hillock to the first myelin internode in resectioned Golgi, immunocytochemical and HRP material showed that the number of synapses varied from 2 to 52 for layers II and III and from 2 to 26 for layer V. The number of synapses on the axon hillocks varied from zero to 12. The variability in these terminations may be an important factor in the shaping of the functional properties of the pyramidal cells.

Animals↗
PubMed1985

The neuronal composition of area 17 of rat visual cortex. III. Numerical considerations.

The neuronal population of area 17 of rat visual cortex has been examined by using tissue from brains fixed by perfusion. The tissue was osmicated and embedded in plastic so that the same neurons could be examined by both light and electron microscopy. In these preparations area 17 was 1.49 mm thick and by stereological procedures it was calculated that there are about 120,000 neurons beneath 1 mm2 of cortical surface. If one assumes area 17 in each hemisphere of the rat to occupy between 7.1 and 9.4 mm2 of cortical surface, then in each hemisphere the area contains between 850,000 and 1,128,000 neurons. Of these neurons 85% are pyramidal cells and 15% are nonpyramidal cells. About one-third of the nonpyramidal cells occur in layers I and VIb, both of which contain only this kind of neuron. The remaining two-thirds of the nonpyramidal cells are in layers II-VIa. Within these layers it has been possible to differentiate bipolar cells from other types of nonpyramidal cells and in each of these two nonpyramidal cell groups to recognize both small and large neurons. The greatest concentration of nonpyramidal cells occurs in layer II/III. To confirm the validity of the stereologically derived data direct counts were made of the medium and large pyramidal cells in layer V.

Animals↗
PubMed1986

Functional organization of crayfish abdominal ganglia: I. The flexor systems.

For insect ganglia, Altman (Advances in Physiological Science, Vol. 23. Neurobiology of Invertebrates. New York: Pergamon Press, pp. 537-555, '81) proposed that individual neuropils control different motor activities. A corollary of this hypothesis is that motor neurons involved in many behavioral functions should branch in more neuropils than those active in fewer behaviors. In crayfish, the abdominal fast-flexor muscles are active only during the generation of the powerstroke for tailflips, whereas the slow-flexor muscles are involved in the maintenance of body posture. The slow flexors are thus active in many of the crayfish's behavioral activities. To test the generality of Altman's idea, we filled groups of crayfish fast-flexor and slow-flexor motor neurons with cobalt chloride and described their shapes with respect to the ganglionic structures through which they pass. Individual fast flexors were also filled intracellularly with HRP. Ganglia containing well-filled neurons were osmicated, embedded in plastic, and sectioned. Unstained sections were examined by light microscopy and pertinent sections were photographed. We found that the paths of the larger neurites were invariant, that the dendritic domains of fast and slow motor neurons occupied distinctive sets of neuropils, and that dendrites of slow motor neurons branched in more ganglionic structures than did those of fast motor neurons. These results are consistent with Altman's hypothesis.

Animals↗
PubMed1991

Glycinergic interplexiform cells make synaptic contact with amacrine cell bodies in goldfish retina.

Recent works utilizing glycine-immunoreactivity (IR) and combined Golgi impregnation and 3H-glycine uptake autoradiography indicate that glycinergic interplexiform cells (IPC) may synapse upon cell bodies in the inner nuclear and ganglion cell layers in fish retina. This possibility was investigated with immunocytochemical techniques using presynaptic and postsynaptic markers for glycinergic neurons: a monoclonal antibody (mAb 7A) against the 93 kDa subunit of the strychnine-sensitive glycine receptor and a polyclonal antiserum against a glycine/BSA conjugate. Synaptic contacts onto the lateral and proximal surfaces of amacrine cell bodies and onto the distal surface of cells in the ganglion cell layer were identified with both probes. The contacts were rare with one contacted amacrine cell/section of 500 linear micron. Serial 1-micron sections were processed alternately for glycine and GABA antisera using postembedding techniques at the light microscopic level. Glycine-IR processes + boutons were apposed to GABA-IR cell bodies in 16 of 17 examples, indicating that the dendro-somatic contacts were onto GABA-immunoreactive amacrine cell bodies. In context of other published morphological data, we suggest that the dendro-somatic synapses were derived from glycinergic IPCs. Glycinergic IPCs receive input from GABAergic horizontal cells and, via a shunt conductance produced by the dendro-somatic contacts, may be involved in controlling the sensitivity, temporal, or spatial properties of amacrine cell responses to large field illumination.

Animals↗
PubMed1992

Survival and integration of bovine chromaffin cells transplanted into rat central nervous system without exogenous trophic factors.

Our previous studies have demonstrated that suspension grafts of isolated bovine chromaffin cells survive in the periaqueductal gray (PAG) of rat midbrain for up to 1 year after transplantation. The current study aimed to determine whether this type of graft could survive transplantation into sites other than the PAG that can benefit from chromaffin cell secretory products. In this study, electron microscope analysis showed that chromaffin cells implanted into the frontal neocortex, striatum, PAG, or the subarachnoid space overlying the spinal cord survived for at least 8 weeks without evidence of degeneration. Intraparenchymally placed grafts appeared relatively avascular and well integrated within the host parenchyma. When blood vessels were found, they were primarily at the host-graft border and were of the nonfenestrated central nervous system (CNS) type. Numerous synapses were present between the grafted cells and host neuronal processes. In addition, extensive intercommunication, via gap junction-like structures, was common in the grafts. Morphologic evidence of granular secretion was most commonly seen in striatal grafts. In contrast, subarachnoid grafts displayed minimal interaction with the host spinal tissue and were heavily vascularized with fenestrated capillaries. Despite morphologic differences between intra- and extraparenchymal grafts, this study demonstrates that isolated suspensions of bovine chromaffin cells survive transplantation into CNS sites without exogenous trophic factors and suggests that these cells are potential candidates for neural transplantation into these regions.

Animals↗
PubMed1992

Unique topographic separation of two spectral classes of cones in the mouse retina.

We have found two immunologically distinguishable cone types in the retina of the mouse, each localized to two opposite halves of the eye. One cone type was labelled by the monoclonal antibody COS-1 specific to the middle-to-long wave sensitive visual pigment of the mammals, while the other type was stained by the shortwave-specific monoclonal antibody (OS-2). These results were confirmed with other antibodies directed against specific sequences of the visual pigments. As a result of the uneven distribution of the two cone types the mouse retina is divided into two fields separated by an oblique meridional line. The middlewave sensitive cones were present exclusively in the dorsal half of the mouse retina (M-field). The overwhelming majority of the shortwave sensitive cones occupied the ventral half (S-field), and only a small number was scattered among the middlewave sensitive cones in the dorsal retina. The ratio of the two cone types in the M-field corresponds to what has been found in the retina of other mammals, including rodents such as the gerbil and the rat. The S-field represents an entirely unique area with the unusually great number of shortwave sensitive cones and with the complete lack of the middlewave sensitive ones. The present study provides the structural basis for dichromacy in a rodent species considered for a long time to be monochromat. In addition, it shows that the ventral retina, containing exclusively S-cones in a relatively high density, is a unique retinal field not present in other mammalian species studied so far.

Animals↗
PubMed1994

Rapid increase in mitochondrial volume in nucleus magnocellularis neurons following cochlea removal.

Second-order auditory neurons in nucleus magnocellularis (NM) of the chick brainstem undergo a series of rapid metabolic changes following unilateral cochlea removal, culminating in the death of 25% of NM neurons. Within hours of cochlea removal, ipsilateral NM neurons show marked increases in histochemical staining for the mitochondrial enzymes succinate dehydrogenase and cytochrome oxidase. We investigated corresponding ultrastructural changes in NM neurons by preparing animals undergoing unilateral cochlea removal for transmission electron microscopy. We quantified changes in NM mitochondrial volume by stereological methods and qualitatively compared mitochondrial morphology between NM neurons destined to survive and those destined to die after cochlea removal. Within hours of cochlea removal, ipsilateral NM neurons show striking increases in mitochondrial volume (84% at 6 hours and 236% at 12 hours after cochlea removal compared to unoperated, control animals). At 2 week survival times, ipsilateral NM neurons contain fewer mitochondria than contralateral neurons. Surprisingly, anesthesia alone causes short-term increases in NM mitochondrial volume. Animals anesthetized with pentobarbital and ketamine and sacrificed 6 or 12 hours later showed a 45% increase in mitochondrial volume compared to previously unanesthetized animals. NM neurons destined to die within days of cochlea removal can be identified within several hours after deafferentation by the appearance of their ribosomes. We observed qualitative differences in mitochondrial morphology in dying neurons. Mitochondria in neurons destined to die consistently showed mitochondrial swelling and vacuolization indicative of metabolic dysfunction. Similar mitochondrial changes have been reported when mitochondria take up excess calcium. Ultrastructural changes in NM after cochlea removal display features of both programmed and pathological cell death, in which increased intracellular calcium is thought to play a role.

Anesthesia↗
PubMed1994

Development of lateral line organs in the axolotl.

Lateral line sensory receptors and their cranial nerves in axolotls arise from a dorsolateral series of placodes, including the octaval placode, that gives rise to the inner ear and the octaval nerve. Anterodorsal and anteroventral placodes occur rostral to the octaval placode and give rise to anterodorsal and anteroventral lateral line nerves and electroreceptors and mechanoreceptors of the snout, cheek, and lower jaw. Middle, supratemporal, and posterior placodes occur caudal to the octaval placode and give rise to similarly named lateral line nerves, electroreceptors and mechanoreceptors of the occipital region of the head, and trunk neuromasts. All placodes, except the posterior placode, elongate, forming sensory ridges, following the genesis of sensory ganglia. Primary mechanoreceptor primordia begin to form within the central zone of the sensory ridges at stage 36; primary electroreceptor primordia originate within the lateral zones of these ridges at stage 38. The first primary mechanoreceptors erupt during stage 37; all primary mechanoreceptors have erupted at hatching (stage 41). Primary electroreceptors begin to erupt at stage 43. Secondary mechanoreceptor primordia begin to form in 1-week-old larvae and erupt 1-2 weeks later. Secondary electroreceptor primordia also begin to form in 1-week-old larvae and continue until clusters of two to five electroreceptors are formed. The developmental stages thought to characterize lateral line placodes in the earliest gnathostomes suggest that this ancestral ontogeny has been truncated in modern amphibians, and ontogenetic mechanisms underlying placodal differentiation are suggested.

Ambystoma mexicanum↗