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Expression of peripherin in solid transplants of foetal spinal cord and dorsal root ganglia grafted to the injured cervical spinal cord of adult rats.

The expression of the neuronal type III intermediate filament protein peripherin was studied in E14 spinal cord fragments and E15 dorsal root ganglia 1-30 weeks after their transplantation to the injured cervical spinal cord of the adult rat. In the dorsal root ganglion transplants, the surviving neurons generally appeared as a rather healthy looking population of small strongly immunoreactive cells which are very similar to the small dorsal root ganglion neurons of adult control rats. In the spinal cord transplants, there were only a few peripherin-immunoreactive neurons, morphologically close to the motoneurons or to the preganglionic sympathetic neurons of adult rats. In both types of transplants, peripherin expression of the immunoreactive neurons was apparently correlated with the previously established ability of these transplanted neurons for extensive axonal growth into a co-grafted peripheral nerve.

Animals↗

Identification of renal sympathetic preganglionic neurons in hamsters using transsynaptic transport of herpes simplex type 1 virus.

Herpes viruses have been used as retrograde transsynaptic tracers to identify pathways from the CNS to specific target tissues. We used herpes simplex virus to identify central nervous system neurons responsible for control of the kidney. Herpes simplex type 1 or herpes simplex type 2 was injected into rat kidneys and herpes simplex type 1 was microinjected into hamster and guinea pig kidneys. After three to seven days, ganglia, spinal cords and brains were examined using immunohistochemistry to visualize the virus-infected neurons. Our first experiments demonstrated that rats were not susceptible to infection with neurotropic strains of herpes simplex type 1. Injections of a wildtype strain of herpes simplex type 2 into rat kidneys led to nonspecific infection of many central nervous system neurons and glia. In contrast, herpes simplex type 1 injections in hamsters and guinea pigs caused specific infection of limited numbers of neurons in approximately one-third of the animals and the study was continued using hamsters. Sympathetic preganglionic neuron labelling was found in the ipsilateral intermediolateral cell column of the spinal cord as well as the lateral funiculus. Most infected preganglionic neurons were located in the seventh to the ninth thoracic spinal segments. Infected neurons were not found in the dorsal or ventral horn of the spinal gray matter and only one or two cells were found in the brainstem. Sympathetic preganglionic neuron morphology was usually normal, showing detailed dendritic arborizations, and lysis was infrequent. Small infected cells were sometimes observed close to sympathetic preganglionic neurons. Because herpes simplex type 1 virus was not detected immunocytochemically in ganglionic neurons in these same hamsters, the polymerase chain reaction was used in some additional hamsters to detect viral DNA in the T12 and T13 chain ganglia and splanchnic ganglia ipsilateral to the kidney injected with herpes simplex type 1. Finally, the overall distribution of renal postganglionic and splanchnic preganglionic neurons in hamsters was examined for comparison to the number and locations of virus-labelled neurons. Retrograde transport of the fluorescent dye FluoroGold demonstrated that (i) renal postganglionic neurons are distributed in the T10-L1 chain ganglia and in the prevertebral splanchnic ganglion and (ii) splanchnic preganglionic neurons are located in the T3-T12 spinal segments, predominantly in the intermediolateral and funicular spinal autonomic nuclei. In conclusion, herpes simplex type 1 virus infected an exclusive population of "renal" neurons in hamsters without lysis and with little cellular reaction to the infection after a survival period of three days, permitting these neurons to be studied in detail.

Animals↗

The location of extrinsic efferent and afferent nerve cell bodies of the normal canine stomach.

The location of the extrinsic efferent and afferent nerve cell bodies to the mucosa, submucosa, and tunica muscularis of the cardiac, gastric, and pyloric gland regions of the ventral stomach and to the mucosa-submucosa alone of these 3 glandular gastric regions was determined using the horseradish peroxidase technique. All animals of the study demonstrated labeling bilaterally in the rostrocaudal extent of the dorsal motor nucleus of the vagus nerve (DMV) although mucosa-submucosa injections resulted in fewer labeled cells in the DMV. There was no evidence of viscerotopic organization within the DMV for the different gastric regions. However, the left nucleus generally contained a greater number of labeled cells than the right nucleus. Injection of the mucosa, submucosa, and tunica muscularis of the cardiac gland region also resulted in labeling in the nucleus ambiguus in 4 of 5 animals. The vast majority of labeled postganglionic sympathetic neurons were found in the celiacomesenteric ganglion. Labeled cells were also located variously in the stellate ganglion, middle cervical ganglion, and sympathetic trunk ganglia for the different groups. There was no discernible pattern of localization of labeled cells within a sympathetic ganglion. For the stomach, afferent labeled cells were located in the range of the first thoracic to fourth lumbar spinal ganglia and the nodose ganglia, bilaterally. As with sympathetic neurons, there was no discernible pattern of localization of labeled cells within a sensory ganglion.

Animals↗

Autoradiographic studies on the distribution of 3H-2,3,4-trimethoxy-beta-phenylethylamine in the mouse.

The regional distribution of radioactivity was studied in the brain and in other organs of the mouse by light microscopic autoradiography after injection of 3H-2,3,4-trimethoxy-beta-phenylethylamine (2,3,4,-TMPEA), a nonhallucinogenic isomer of mescaline. The distribution patterns were compared with biochemical and autoradiographic results obtained after 3H-mescaline. Although 2,3,4-TMPEA is rapidly deaminated compared to mescaline its distribution pattern in the brain is similar to that of mescaline at 1 h after injection. However, contrary to mescaline the radioactive labeling becomes more and more homogenously distributed in the brain with tiem. The distribution patterns in kidney, pancreas, adrenal, and spinal ganglia were also different from those observed after application of 3H-mescaline.

Animals↗

Brain cooling in humans--anatomical considerations.

Vascular arrangements allowing a bulky transfer of venous blood from the skin of the head and from nasal and paranasal mucous membranes to the dura matter provide an excellent anatomical basis for the convection process of cooling, caused by evaporation of sweat or mucus. The dura mater, with its extraordinarily high vascularization controlled by a potent vasomotor apparatus, may transmit temperature changes to the cerebrospinal fluid (CSF) compartment. Temperature gradients of the CSF may in turn influence the temperature of brain parenchyma (1) directly, along the extensive contact area between the cerebrocortical surface and the CSF-compartment, or (2) indirectly, via brain arteries that extend over long distances and arborize within the subarachnoid space before entering the pial vascular network and brain parenchyma. Numerous subarachnoid and pial arterial branches exposed to the CSF have diameters in the range of the vessels of the retia mirabilia of animals in which selective brain cooling has been clearly established experimentally. It is also shown that the arrangements of venous plexuses within the vertebral canal provide anatomical preconditions for a cooling of the spinal cord via the CSF. The possibility of spinal cord and spinal ganglia cooling by temperature convection via venous blood--cooled in the venous networks of the skin of the back--flowing through numerous anastomoses to the external and internal vertebral plexuses and, finally, into the vascular bed of the spinal dura is discussed on the basis of anatomical facts.

Arachnoid↗

Afferent projections from the rat longus capitis muscle studied by transganglionic transport of HRP.

Using the method of transganglionic transport of horseradish peroxidase (HRP) according to Mesulam (1978), the course and central terminations of the afferents from the longus capitis muscle were studied in the rat. Application of HRP to the cut muscle nerves was followed by heavy labeling of a considerable number of cell bodies of different sizes in the spinal ganglia C2 and C3. The labeled central processes follow two distinct main routes: one to the external cuneate nucleus, which is known to project ipsilaterally to the cerebellum, the other to the central cervical nucleus (CCN) of the spinal cord. The latter nucleus contains dense arborizations and terminals of muscle afferents in close relationship to medium sized cells which project contralaterally to the cerebellum. This could be shown in double labeling experiments with HRP as a tracer for primary afferents and Nuclear yellow for the cerebellar pathway. The labeled area of CCN extends from the fourth cervical segment up to the medulla oblongata where it lies laterally adjacent to the hypoglossal nucleus, though clearly separated from the latter. In the cervical part of CCN, dendrites of neck muscle motoneurons arborize within the area of afferent terminals. Besides the external cuneate and the central cervical nuclei, afferent projections were seen in lamina X, partly in close contact with the central canal, and in more lateral areas of lamina VII. In contrast to results from studies on other neck muscles, no reaction product was observed in dorsal horn laminae I-VI.

Afferent Pathways↗

Involvement of substance P neurons in contractions of canine small intestine produced by mesenteric nerve stimulation.

Pathways for contractions of in vivo canine small intestine produced by mesenteric nerve stimulation (MNS) were studied. In intact and chronically sympathectomized dogs, contractions of jejunal and ileal segments were largely reduced by intra-arterial infusion of capsaicin (10-100 microM, 0.07 ml/min), substance P (SP) antagonist, (D-Pro4, D-Trp7.9) SP (4-11) (100 microM, 0.14 ml/min), hexamethonium (100-1000 microM, 0.07 ml/min) or atropine (100 microM, 0.07 ml/min). In chronically vagotomized dogs, capsaicin, SP-antagonist or atropine significantly reduced MNS-induced contractions, but hexamethonium did not. In dogs in which the coeliac and superior mesenteric ganglia had previously been removed, MNS caused no response although intra-arterial injection of 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP, 0.1 mumol) caused marked contractions. It may therefore be suggested that extrinsic SP neurons probably originating in spinal ganglia and intrinsic SP neurons receiving input from vagal preganglionic cholinergic neurons are involved in the excitatory pathways to MNS-induced contractions and that activation of these neurons excites myenteric cholinergic neurons, thereby causing contractions of the small intestine.

Adrenergic Fibers↗

The formation of mesoderm and mesectoderm in 5- to 41-somite rat embryos cultured in vitro, using WGA-Au as a marker.

The formation of mesectodermal cells by the neural crest in 5- to 41-somite stage embryos was investigated experimentally in rat embryos cultured in vitro, using lectin-coated colloidal gold as a probe. This method labelled all ectodermal cells, among them neural crest, surface ectodermal placodal and epiblastic (primitive streak) cells. The neural crest provides the mesodermal compartment of the entire head region with cells, including the primitive cranial ganglia and the branchial arches. In the head region migration of neural crest cells over a great distance (long-distance migration) was not observed. In the trunk region neural crest derived cells were mainly found to form the primitive spinal ganglia and the sympathetic trunk, once again without long-distance cell migration. Structures and tissues that supposedly were derived from the primitive streak were hardly labelled with colloidal gold. Surface ectodermal placodes were not only found at the expected sites (e.g. epibranchial placodes) but also in the ectoderm covering the transverse septum and lateral abdominal walls.

Animals↗

Innervation of the posterior fossa dura of the cat.

This study was designed to identify the location of neurons giving rise to fibers innervating the posterior fossa dura in the cat using horseradish peroxidase (HRP, Sigma, Type VI). Investigations since the 19th century have implicated innervation by cranial nerves V, VII, IX, X and XII and the upper cervical nerves, C1-3. The meninges of the posterior fossa of 14 cats was exposed using one of three surgical approaches: (1) a suboccipital craniectomy and C1 laminectomy, (2) a parieto-occipital craniectomy with removal of the occipital lobe and bony tentorium exposing the meninges over the cerebellum, or (3) an anterior approach through the upper neck, exposing the dura of the ventral surface of the caudal brainstem. A unilateral, curvilinear incision was made in the dura and HRP was applied to the exposed dural edges. Following 48 hours the animals were sacrificed and fixed by perfusion. Cranial nerve ganglia of V, VII, IX, X, dorsal root ganglia (DRG) of C1-3, and superior cervical ganglia (SCG) were removed bilaterally, sectioned and processed with tetramethylbenzidine (TMB). HRP labeled cells were located bilaterally, always more ipsilaterally, in DRG of C1, C2, C3, and SCG with application of HRP to all three regions of the dura. Labeled cells were also located in trigeminal ganglia and superior ganglia of CN X, occasionally bilaterally, depending on the site of application. No HRP was ever identified in neurons of the geniculate ganglion, inferior ganglion of CN X or superior or inferior ganglia of CN IX. This information is valuable to an understanding of the innervation of intracranial structures and the problems of head pain.

Afferent Pathways↗

Herpes simplex types 1 and 2: latency in the genital tract of guinea pigs.

Guinea pigs were infected with herpes simplex virus (HSV) intravaginally and then sacrificed during latent infection. Virus was recovered from the ganglia, spinal cord and genital tissues by co-cultivation after 1-6 weeks in culture. The virus could not be recovered from the genital tract during the first week of co-cultivation, nor from homogenized genital tissue. Cultivation of genital tissues with acyclovir did not reduce the recovery of HSV. Thus, HSV appeared to establish a truly latent infection in the genital tract and not a persistent infection as previously described.

Acyclovir↗

Distribution and development of VIP immunoreactive neurons in the spinal cord of the embryonic and newly hatched chick.

The distribution and development of vasoactive intestinal polypeptide (VIP) immunoreactive elements were studied in the spinal cord of embryonic and newly hatched chicks with the indirect immunofluorescence method. VIP neurons were first detectable in the presumed dorsal horn at stages 27-28 (incubation day 5). Subsequently they increased in number, and by stage 39 (day 12) many occurred in lamina I, in the nucleus of the dorsolateral funiculus, and in the lateral portion of the neck of the dorsal horn throughout the cord. However, at the thoracic level many were also situated lateral to the central canal, with their processes running to the ipsilateral lateral and contralateral ventral funiculi. The pattern described above remained visible in both embryonic and colchicine-pretreated newly hatched chicks. During development, VIP fibers appeared later than cell bodies. In the gray matter, they were mainly scattered in the intermediate zone, especially around the central canal at all levels examined. In the white matter, however, longitudinal fibers were observed in the lateral funiculus throughout the cord, but mostly at the cervical level, though some also occurred in the ventral funiculus. This finding supports the idea that spinal VIP neurons might project rostrally via the lateral funiculus. In addition, no VIP immunoreactivity was found in the spinal ganglia, but examination of the sympathetic paravertebral ganglia showed immunoreactivity as described by others.

Age Factors↗

Actin myopathy with nemaline bodies, intranuclear rods, and a heterozygous mutation in ACTA1 (Asp154Asn).

Mutations in the skeletal muscle alpha-actin gene ( ACTA1) are associated by and large with three muscle diseases (1) congenital actin myopathy, (2) nemaline myopathy, and (3) intranuclear rod myopathy. More than 70 mutations have now been identified. The majority of ACTA1 mutations are dominant, a small number are recessive and most isolated cases with no previous family history have de novo dominant mutations. The present case, a boy of healthy Turkish parents, had a severe form of the disease of the latter type due to a heterozygous, presumably de novo mutation of the ACTA1 gene in exon 4 (Asp154Asn), with lack of spontaneous movements at birth requiring immediate mechanical ventilation. He died at the age of 9 weeks due to respiratory failure, secondary pneumonia, and chylothorax. The biopsy specimen of the femoral muscle was characterized by pleomorphic alterations with numerous muscle fibers showing accumulation of actin filaments, but, in addition, both nemaline bodies and intranuclear rod bodies. This was also seen in several other muscles investigated at autopsy. No developmental abnormalities of the central nervous system, and no loss of spinal motor neurons were detected despite atrophy or hypotrophy of a considerable number of muscle fibers. The peripheral nervous system, which has not been studied before in patients with ACTA1 mutations, showed no loss of motor or sensory myelinated fibers and no loss of sensory neurons in spinal ganglia.

Actins↗

Age-related mitochondrial damage in the B-type cells of the rat trigeminal ganglia.

Aging is associated with signs of sensory impairment and neurological symptoms. Advancing age is characterized by increased thresholds of thermal, tactile and vibratory sensations. One important cause of the sensory disturbances has been stated to be the loss of neurons. Decreases have been observed in the number of peripheral nerve fibers and in the number of neurons in the spinal ganglia of rats. In the present study, the cytoplasmic organelles of the neurons of the trigeminal ganglia were examined in young and senescent rats in order to reveal the cause of cell loss during aging. Mitochondrial alterations, swelling and loss of internal cristae were observed from 23 week of age in the B-type neurons of the trigeminal ganglia. Other cytoplasmic elements were intact. Mitochondrial damage was never seen in A-type neurons and satellite glial cells. It was concluded that the ultrastructural changes in the mitochondria of the B-type cells may contribute to the nervous disturbances that occur in senescent individuals. The diminution of mitochondrial damage and the protection of B-type neurons through the use of nerve growth factors may prevent the sensory impairment late in life.

Aging↗

A modified roller tube technique for organotypic cocultures of embryonic rat spinal cord, sensory ganglia and skeletal muscle.

The roller tube technique as initially described in the literature in 1981, was modified in several aspects for the coexplantation of embryonic rat spinal cord with attached dorsal root ganglia and skeletal muscle from newborn rats. The high metabolic activity of this coculture system required a particular culturing protocol to stabilize pH and osmotic pressure. The appropriate adjustment of the partial pressure of carbon dioxide gas in the incubator proved to be essential for the control of the pH within narrow limits (7.3 +/- 0.1). The adjustment of the osmotic pressure of the medium (290-300 mOsm) improved the growth of the cultures considerably. Roller drum speed was set to 120 revolutions per hour for enhanced flattening of the culture. A simple rating system was used to evaluate neuronal and non-neuronal outgrowth under different modifications of the culture system. Furthermore, morphological and electrophysiological criteria were defined for evaluating individual neurons. The technique described insures the growth of long-term organotypic cocultures of spinal cord, sensory ganglia and skeletal muscle.

Animals↗

[Efferent sympathetic preganglionic and afferent spinal transit pathways of the cat stellate ganglion].

Using a technique of retrograde axonal transport of horseradish peroxidase, labeled neurons were detected in the intermedialateral nucleus (pars principalis and pars funicularis), intercalatous spinal nucleus, and in the ventral horns of the spinal cord in cats. Afferent spinal transit pathways pass in all the above branches as well as the vertebral nerve. Bodies of the labeled neurons with branches passing in the vertebral nerve are located in the T2-T7 spinal ganglia, whereas those with branches passing in other nerves--are located in the C8-T8.

Afferent Pathways↗

Atrial natriuretic factor-like immunoreactivity in spinal cord and in primary sensory neurons of spinal and trigeminal ganglia of guinea-pig: correlation with tachykinin immunoreactivity.

Atrial natriuretic factor (ANF) is a cardiac hormone with various functions in body homeostasis. It is also processed in the brain and in the peripheral nervous system where it appears to play a role as a neuromodulator. Little is known about the presence of ANF throughout the spinal cord of the guinea-pig. We therefore examined the distribution of ANF and its possible interrelation with primary sensory afferents in this species. Using enzyme-and fluorescence-immunohistochemistry on deparaffinized sections. ANF-like immunoreactivity was found to be present in nerve fibers in laminae I/II of the spinal cord and in neurons of spinal and trigeminal ganglia. Tachykinins and ANF coexisted in very few fibers of the spinal cord but did not coexist in primary sensory spinal or trigeminal neurons. Our results indicate that spinal ANF-immunoreactive fibers are of dual origin, primary sensory and non-primary sensory. The possibly heterogeneous source of the non-primary sensory ANF, its possible coexistence with other co-transmitters and functional implications are discussed.

Animals↗