Localization of chick retinal 24,000 dalton protein (visinin)-like immunoreactivity in the rat lower brain stem

The distribution of visinin, a 24,000 dalton peptide, in the lower brain stem of the rat was examined by means of an indirect immunofluorescent method. Visinin-immunoreactive structures were found to be unevenly distributed only in the neuronal elements. The following neuronal systems were strongly labeled by the antiserum; the Purkinje cell system, mammillotegmental system, habenulointerpeduncular system, the second layer of the superior colliculus, ventral tegmental area, substantia nigra pars lateralis, area medial to the medial geniculate body, parabrachial area, dorsal and ventral nuclei of the lateral lemniscus, pontine reticular formation just medial to the trigeminal principal nucleus, superior olivary nucleus, solitarii nucleus, external layer of the inferior colliculus and spinal trigeminal nucleus. The densities of the labeled fibers in these areas paralleled those of the labeled cells. In addition, highly dense visinin-immunoreactive fiber plexuses were seen in the zona compacta of the substantia nigra, lateral portion of the interpeduncular nucleus, ventral tegmental nucleus of Gudden and vestibular nucleus.     

Calcitonin gene-related peptidergic projection from the parabrachial area to the forebrain and diencephalon in the rat: an immunohistochemical analysis

We investigated ascending fiber projections of calcitonin gene-related peptide from the parabrachial area to the forebrain and diencephalon in the rat using immunocytochemistry. Destruction of the lateral portion of the dorsal parabrachial area resulted in a marked ipsilateral decrease in the fibers containing calcitonin gene-related peptide in the ventromedial hypothalamic nucleus, indicating that cells containing calcitonin gene-related peptide in the lateral portion of the dorsal parabrachial area projected to the ipsilateral ventromedial hypothalamic nucleus. Destruction of the ventral portion of the parabrachial area resulted in a marked decrease of fibers containing calcitonin gene-related peptide in the bed nucleus of the stria terminalis, the central amygdaloid nucleus and the lateral hypothalamus just medial to the crus cerebri (the far-lateral hypothalamus), and a less marked decrease in the ventromedial thalamic nucleus. This means that there are projections from cells containing calcitonin gene-related peptide in the ventral portion of the parabrachial area to the first three regions just mentioned, and to some extent to the last.     

Correlation between met-enkephalin and substance P immunoreactivity in the primate globus pallidus

Using immunohistochemical techniques, the distribution of enkephalin and substance P immunoreactive fibers and terminals was studied in the globus pallidus of the non-human primate. In the external segment of the globus pallidus, enkephalin immunoreactivity was very dense while only sparse to moderate substance P staining was observed. Enkephalin immunoreactivity in the inner portion of the internal segment was moderate while such fibers were sparse in the outer portion of the internal segment. Substance P immunoreactivity was dense throughout the internal segment of the globus pallidus.The pattern of enkephalin and substance P immunoreactivity in the globus pallidus of the non-human primate as reported in the present study is of interest with regard to pallidal efferents. The pallidosubthalmic projection, arising from the external segment of the globus pallidus, is likely to be strongly influenced by the very dense network of enkephalin immunoreactive fibers and terminals in this region. Conversely, substance P immunoreactive elements are sparse in the external segment, and are, therefore, unlikely to influence significantly activity carried by the pallidosubthalmic projection. Since the inner portion of the internal segment contains moderate enkephalin immunoreactivity and dense substance P immunoreactivity, the information carried by the lenticular fasiculus may be modulated by both of these putative transmitters. On the other hand, based on the densities of immunoreactivity, the ansa lenticularis, which arises from the outer portion of the internal segment, is likely to be under greater influence of the dense substance P projection to this area.     

Oxytocin/vasopressin-like immunoreactivity is present in the nervous system of hydra

Nerve cells have been found in hydra, which react with antisera to oxytocin, vasopressin and mesotocin. These nerve cells have a high density in the ectoderm of basal disk and tentacles and lower density in the ectoderm of peduncle, gastric region and hypostome. A very small number of nerve cells occur also in the endoderm of foot, gastric region and hypostome. By using a technique for simultaneous visualisation of nerve cells reacting with antisera to oxytocin and vasopressin, it can be shown that these nerve cells belong to a single population. In agreement with this, the staining of the nerve cells can be abolished by absorbing each antiserum with either oxytocin, vasopressin, [Lys⁸]vasopressin, vasotocin, mesotocin or isotocin, indicating that the antigenic determinant of hydra cross-reacts with those antibody subpopulations, which recognize common portions (sequence 1–2, 5–7, 9) of the oxytocin/vasopressin-like peptides. With radioimmunoassays that are specific for either oxytocin or vasopressin, only very low amounts of immunoreactivity were measured. In addition, the dilution curves in these assays were not parallel to the standards, indicating that the antigenic determinant of hydra is not oxytocin or vasopressin.The presence of oxytocin/vasopressin-like material in coelenterates, shows that this family of peptides is of great antiquity.     

Distribution of galanin immunoreactivity in the central nervous system and the responses of galanin-containing neuronal pathways to injury

Radioimmunoassay and immunocytochemistry were used to study the distribution of galanin, a novel 29 amino acid porcine intestinal peptide, in the central nervous system of the rat and pig. The pattern of distribution was similar in the two species, with the highest concentrations of galanin-like immunoreactivity found in the neurohypophysis, hypothalamus and sacral spinal cord. Immunocytochemical studies of these regions localized galanin-like immunoreactivity to cell bodies in the paraventricular and supraoptic nuclei of the hypothalamus, to fibres in the pars nervosa and to numerous cell bodies and fibres in the dorsal horn of the spinal cord. On both gel and high pressure liquid chromatography, galanin-like immunoreactivity in rat and pig nervous tissue eluted as a single peak in a position similar to purified procine intestinal galanin standard. Surgical and pharmacological manipulations in the rat suggest the presence of galanin in afferent fibres. An increase of galanin-like immunoreactivity was observed in the sacral spinal cord of the rat following thoracic spinal cord transection. Thus galanin-like immunoreactivity in the brain is mainly localized in the hypothalamopituitary region. The decrease of galanin-like immunoreactivity in the dorsal horn of the spinal cord, following dorsal rhizotomy and pre-treatment of rats with capsaicin, indicates that many of the fibres, which are of small diameter, may well be derived from spinal sensory neurones.     

Immunohistochemical localization of protein-O-carboxylmethyltransferase in rat brain neurons

The distribution of the enzyme protein-O-carboxylmethyltransferase (EC 2.1.1.24) has been investigated in the rat brain using both immunohistochemical and biochemical techniques. The enzyme, which carboxylmethylates free aspartic and glutamic acid residues of protein substrates, was localized in neurons, but not other cell types throughout the brain. The highest immunoreactivity was detected throughout the cortex, followed by the hippocampus, the corpus striatum, the thalamus and the amygdala. Immunoreactive cells were detected in other brain regions but were not as prominent as those regions listed above. The distribution of immunoreactivity in the hippocampus was most striking, with considerable labelling of the pyramidal and granule cells in all regions. Numerous pyramidal cells were labelled in the cerebral cortex, with some ascending processes exhibiting immunoreactivity. The corpus striatum was uniformly labelled, suggesting that the enzyme was not localized to any specific neurotransmitter system. The antisera employed in this study was generated against purified bovine brain protein-O-carboxylmethyltransferase and Western immunoblot analysis showed cross reactivity against both rat brain and human erythrocyte forms of the enzyme. Enzyme activity and methyl acceptor protein capacity were examined in 1.5 mm coronal sections of rat brain. The regions with highest enzyme activities were found in cross-sections containing cortex and corpus striatum or cortex and hippocampus. The lowest enzyme activities were noted in slices of brainstem and cerebellum, areas exhibiting low amounts of immunoreactive protein-O-carboxylmethyltransferase. Methyl acceptor protein capacity was highest in slices of cortex and corpus striatum, cortex and hippocampus and was lowest in slices of brainstem and cerebellum. These results demonstrate that protein-O-carboxylmethyltransferase has an unique neuronal pattern of distribution in the rodent central nervous system, and suggest that the carboxylmethylation of proteins may be of functional significance in these neurons.     

An immunohistochemical study of the projections of somatostatin-containing neurons in the guinea-pig intestine

Somatostatin-like immunoreactivity was localized in nerves in whole mount preparations of the separated layers of the guinea-pig intestine. The directions in which the neurons project were determined by examining the accumulation of somatostatin-like immunoreactivity after axonal flow was interrupted. In some experiments this was done by crushing or cutting the nerves in isolated preparations which were then maintained in oxygenated Krebs solution for 3–5 h. In other experiments, the nerves were cut in vivo and the animals allowed to survive for 4–8 days before the intestine was examined.Somatostatin immunoreactive nerve cell bodies were found in both the myenteric plexus, where they represented 4.7% of the total population of neurons, and in the submucous plexus, where they formed 17.4% of the total population. The axons of the somatostatin-containing neurons in the submucosa are not polarized while those of the somatostatin-containing neurons in the myenteric plexus of the small intestine project in the anal direction for 8–12 mm to form pericellular baskets around other enteric neurons, some of which are reactive for somatostatin.It is postulated that somatostatin-containing neurons in the myenteric plexus are interneurons in a descending nerve pathway, possibly the one involved in the descending inhibitory reflex of peristalsis.     

Somatostatin-like immunoreactivity in the forebrain of Pseudemys turtles

Previous investigations of cortical organization in the brain of the turtle have revealed many features typical of mammalian neocortex. Recent evidence suggests that many neocortical neurons contain neuroactive peptides. The possibility that one such peptide, somatostatin, is found in the turtle brain was tested using immunocytochemical techniques. Intense somatostatin-like immunoreactivity was observed in many neurons and fibers in turtle cortex, as well as in several forebrain nuclei. Cortical neurons with several different dendritic configurations showed immunoreactive labelling, including bipolar, stellate and pyramidal cell types. In addition, stained cells and processes were observed in close association with the ependyma of the lateral ventricle. Other forebrain regions containing immunoreactive neurons included the dorsal ventricular ridge, the basal telencephalic nuclei and the hypothalamus. These data support the idea that peptidergic neurons existed in the pallium of an ancestor common to modern mammals and reptiles. We speculate that somatostatin plays a similar role in the normal function of all types of cortex and suggest that turtle cortex may provide a useful model for the study of this cortical neuropeptide.     

Pancreatic polypeptide immunoreactivity in rat brain is actually neuropeptide Y

Radioimmunoassay was combined with high pressure liquid chromatography and immunohistochemistry to establish the identity of pancreatic polypeptide-like immunoreactive material in the central nervous system of the rat. Antisera to avian pancreatic polypeptide, bovine pancreatic polypeptide, the invariant amidated carboxyterminal hexapeptide fragment of mammalian pancreatic polypeptides and the structurally related peptide, neuropeptide Y, were used immunocytochemically to localize neurons containing immunoreactive pancreatic polypeptide-like material in rat brain. Adjacent brain sections stained by the indirect immunofluorescent technique and single sections from double-staining experiments demonstrated that identical fibers and perikarya stained for pancreatic polypeptide-like immunoreactive material by antisera directed against each of the four peptides. Characterization of pancreatic polypeptide-like immunoreactive material in chromatographed rat brain extracts by radioimmunoassay using antisera to either neuropeptide Y or the carboxy-terminal portion of the pancreatic polypeptide molecule revealed that the major peak of immunoreactive material, as measured by either assay, appeared to co-elute with synthetic porcine neuropeptide Y. A minor peak of immunoreactive material co-eluting with peptide YY standard was indicated by the neuropeptide Y radioimmunoassay. This was contrasted by data obtained from chromatographic profiles of rat pancreas, which showed that the main immunoreactive peak, as measured by the neuropeptide Y assay, co-eluted with porcine peptide YY, with a minor peak co-eluting with porcine neuropeptide Y. The main peak of immunoreactive material in pancreas, as measured by the pancreatic polypeptide carboxy-terminal radioimmunoassay, eluted considerably earlier than standard peptide YY, neuropeptide Y and bovine pancreatic polypeptide, and is probably identical to rat pancreatic polypeptide.(ABSTRACT TRUNCATED AT 250 WORDS)     

Overall distribution of vasoactive intestinal polypeptide-containing nerves on the wall of cerebral arteries: an immunohistochemical study using whole-mounts

The overall distribution of vasoactive intestinal polypeptide (VIP)-like immunoreactivity on the wall of the cerebral arteries, including its 3-dimensional profile, was investigated by means of the indirect immunofluorescence method using flat-mounts. VIP-immunoreactive fibers run spirally on the wall of the cerebral arteries. On the wall of the large arteries, such as the vertebral artery, basilar artery, internal carotid artery, within and/or without the circle of Willis, posterior and anterior communicating arteries, proximal parts of anterior, mid and posterior cerebral arteries, these fibers are richly distributed and show a dense grid-like appearance. The highest density was identified on the wall of the anterior cerebral artery, internal carotid artery and anterior communicating artery, while the lowest density was on the posterior communicating artery. On the other hand, on the walls of the branches of these arteries or along distal parts of the anterior, mid and posterior cerebral arteries, the number of VIP-immunoreactive fibers decreased markedly.     

Distribution of atrial natriuretic factor-like immunoreactive neurons in the rat brain

Using antisera generated in rabbits against rat atriopeptin III [alpha-rANP(5-28)] and human alpha-atrial natriuretic polypeptide we mapped the distribution of atrial natriuretic factor-like immunoreactivity throughout the rat central nervous system. Cell bodies were observed in the telencephalon (nucleus interstitialis striae terminalis and between the amygdala centralis and medialis), throughout the diencephalon in all nuclei of the "anteroventral third ventricle", the base of the hypothalamus, the subzona incerta area, the medial forebrain bundle and the medial habenula, in the mesencephalon (mamillary body, substantia nigra lateralis, dorsal and ventral parabrachial nuclei) and very sparse in the medulla oblongata along the fourth ventricle towards the vestibular nuclei, the nucleus tractus solitarii and nervi trigemini. Fibers were present wherever cell bodies were located. The highest relative densities were observed in the anteroventral third ventricle area and the medial habenula. Sparse fibers were also seen in the spinal cord (dorsal and ventral horn and around the central canal) and in the posterior pituitary. The predominance of the atrial natriuretic factor-like perikarya and fibers in the anteroventral third ventricle area suggests an involvement of this peptide in central blood pressure control.     

Isolation and partial characterisation of neuronal growth cones from neonatal rat forebrain

We have devised a method for the isolation of viable neuronal growth cones from neonatal rat forebrain. The method involves differential and density gradient centrifugation and exploits the relatively low buoyant density (approximately 1.018 g/cm3) of growth cones. There are no known biochemical markers for growth cones and it was necessary therefore to monitor for their presence during the isolation using transmission electron microscopy. Several criteria were used to identify isolated growth cones including the presence of filopodia, an extensive system of branching, tubular smooth endoplasmic reticulum and a region rich in microfilaments subjacent to the plasma membrane. These morphological features are similar to those of growth cones identified unequivocally in intact developing brain and in tissue culture. Electron microscopical analysis showed that greater than 90% of membrane-bound, identifiable objects in one fraction were growth cones by these criteria. The major contaminant consisted of membrane sacs and vesicles of unidentified origin. There were only small amounts of isolated rough endoplasmic reticulum and mitochondria. Isolated growth cones were roughly spherical in shape with a diameter of 1.9 +/- 0.5 micron (mean +/- 1 SD). They usually contained mitochondria, large granular vesicles and small vesicles, and occasionally contained coated vesicles, lysosomes, lamellar bodies and multivesicular bodies, and only very rarely, intermediate filaments. Occasionally, growth cones had rudimentary synapses on them. The viability of isolated growth cones was investigated by observing their behaviour in short-term culture. After a few hours in culture on poly-D-lysine-coated coverslips, growth cones flattened down and extended filopodia-like processes. This behaviour was inhibited by cytochalasin B and reversibly by cold (4 degrees C). We conclude that physiologically active growth cones can be isolated rapidly and in large numbers by the method described here.     

Perivascular nerves with immunoreactivity to vasoactive intestinal polypeptide in cephalic arteries of the cat: distribution, possible origins and functional implications

The distribution of nerves containing vasoactive intestinal polypeptide(VIP)-immunoreactive material was examined in the cephalic arteries and cranial nerves of cats using an indirect immunofluorescence procedure on whole mounts. Perivascular VIP-immunoreactive nerves were widely distributed in arteries and arterioles supplying glands, muscles and mucous membranes of the face. Within the cerebral circulation, perivascular VIP-immunoreactive nerves were most abundant in the circle of Willis and the proximal portions of the major cerebral arteries and their proximal branches supplying the rostral brainstem and ventral areas of the cerebral cortex. Nerves containing VIP-immunoreactive material were absent from distal portions of arteries supplying the posterior brainstem, cerebellum and dorsal cerebral cortex. Cerebral perivascular VIP-immunoreactive nerves had extracerebral origins probably from VIP-immunoreactive perikarya within microganglia in the cavernous plexus and external rete. Extracerebral perivascular VIP-immunoreactive nerves probably arose from VIP-immunoreactive perikarya in microganglia associated with the tympanic plexus, chorda tympani, lingual nerve and Vidian nerve as well as from cells in the otic, sphenopalatine, submandibular and sublingual ganglia. Therefore, it seems likely that each major segment of the cephalic circulation is supplied by local VIP-immunoreactive neurons. If the VIP-immunoreactive nerves cause vasodilation, they are well placed to allow redistribution of arterial blood flow within the head. During heat stress, neurogenic vasodilation of the appropriate beds would permit efficient cooling of cerebral blood, particularly that supplying the rostral brainstem and surrounding areas of the cerebral cortex.     

Distribution of calcitonin gene-related peptide in the rat peripheral nervous system with reference to its coexistence with substance P

This immunocytochemical study, using a double-staining method, showed that calcitonin gene-related peptide-like immunoreactive structures are widely distributed in the peripheral nervous system and that many of them coexist with substance P-like immunoreactive structures in single sensory ganglion cells. Neurons positive for calcitonin gene-related peptide but negative for substance P were detected in sensory ganglia. These cells were large (about 30-45 micron in diameter); these primary sensory neurons containing calcitonin gene-related peptide can probably act independently of substance P. There were neurons containing calcitonin gene-related peptide without substance P in the pterygopalatine ganglion, although these cells were less numerous than in the sensory ganglia. In consecutive sections, calcitonin gene-related peptide-like structures occurred in thyroid parafollicular cells, which also contain calcitonin. This suggested that messenger RNA for producing calcitonin gene-related peptide is also present in the thyroid, and like calcitonin, calcitonin gene-related peptide may have a peripheral physiological role.     

Immunohistochemical distribution of somatostatin-like immunoreactivity in the central nervous system of the adult rat

The localization and distribution of somatostatin (growth hormone release-inhibiting hormone; somatotropin release-inhibiting factor) have been studied with the indirect immunofluorescence technique of Coons and collaborators and the immunoperoxidase method of Sternberger and coworkers using specific and well-characterized antibodies to somatostatin, providing semiquantitative, detailed maps of somatostatin-immunoreactive cell profiles and fibers. Our results demonstrate a widespread occurrence of somatostatin-positive nerve cell bodies and fibers throughout the central nervous system of adult, normal or colchicine-treated, albino rats. The somatostatin cell bodies varied in size from below 10 micron up to 40 micron in diameter and could have only a few or multiple processes. Dense populations of cell somata were present in many major areas including neocortex, piriform cortex, hippocampus, amygdaloid complex, nucleus caudatus, nucleus accumbens, anterior periventricular hypothalamic area, ventromedial hypothalamic nucleus, nucleus arcuatus, medial to and within the lateral lemniscus, pontine reticular nuclei, nucleus cochlearis dorsalis and immediately dorsal to the nucleus tractus solitarii. Extensive networks of nerve fibers of varying densities were also found in most areas and nuclei of the central nervous system. Both varicose fibers as well as dot- or "dust-like" structures were seen. Areas with dense or very dense networks included nucleus accumbens, nucleus caudatus, nucleus amygdaloideus centralis, most parts of the hypothalamus, nucleus parabrachialis, nucleus tractus solitarii, nucleus ambiguus, nucleus tractus spinalis nervi trigemini and the dorsal horn of the spinal cord. One exception is the cerebellum which only contained few somatostatin-positive cell bodies and nerve fibers. It should be noted that somatostatin-positive cell bodies and fibers did not always conform to the boundaries of the classical neuroanatomical nuclei, but could often be found in areas between these well-established nuclei or occupying, in varying concentrations, only parts of such nuclei. It was difficult to identify with certainty somatostatin-immunoreactive axons in the animals studied. Some pathways could, however, be demonstrated, but further experimental studies are necessary to elucidate the exact projections of the somatostatin-immunoreactive neurons in the rat central nervous system.     

Light and electron microscopic immunocytochemical localization of adrenocorticotrophin-like activity in rat cerebellar and red nuclei

An antiserum raised to synthetic adrenocorticotrophin (ACTH) was bound to large neurons of the cerebellar nuclei in rat. In these neurons, the matrix microtubules of the cell bodies and dendritic processes were ACTH-positive. In the axon terminals, 40 nm diameter clear synaptic vesicles were stained in both the deep cerebellar nuclei and red nucleus. Following transection of the superior cerebellar peduncle, ACTH-labeled terminals disappeared in the red nucleus, suggesting that the ACTH-labeled neurons were projection neurons of the cerebellar nuclei.     

Somatostatin-like immunoreactivity in amacrine cells of the chicken retina

Somatostatin-like immunoreactivity was detected in chicken retina by radioimmunoassay. The levels of somatostatin-like immunoreactivity decreased after intra-ocular injection of kainic acid, but were not affected by destruction of the ganglion cells. By immunohistochemistry, somatostatinimmunoreactive amacrine cells were found in the inner nuclear layer. These cells were destroyed by kainic acid. At least some of the cells projected to all three sub-layers of the inner plexiform layer in which there were diffuse bands of fluorescence. Specific immunofluorescence was also detected at the level of the outer limiting membrane and the optic nerve fibre layer, but the outer nuclear and plexiform layers, horizontal, bipolar and ganglion cells did not show specific immunofluorescence.It is suggested that other amacrine cell sub-classes, defined in terms of their putative transmitter, may show specific patterns of cell body location and size, and terminal arborisation.     

Treatment of sections of chick retina with acetylcholinesterase increases the enkephalin and substance P immunoreactivity

Frozen sections 10 microns thick were cut from the retina of chicks which had been kept either in total darkness or in a well lit room. The sections were incubated with acetylcholinesterase before antibodies to [Leu] enkephalin, substance P or somatostatin were applied. Sections of bovine adrenals were treated similarly but they were developed only with antibodies to [Leu]enkephalin. There were low numbers of immunoreactive amacrine cells and processes when any of the three antibodies were used on sections of dark-adapted retinae. When the sections were treated with acetylcholinesterase, however, the enkephalin-like and substance P-like immunoreactivity was enhanced while there was no effect on somatostatin. Counts of immunofluorescent cells indicated that the numbers had increased to levels like those found in light-adapted retinae. The adrenal also showed an enhanced enkephalin-like immunoreaction after treatment with the enzyme. Incubation with buffer alone or with enzyme together with 10 mM acetylcholine abolished the reaction. Acetylcholinesterase treatment of sections from light-adapted retinae had no discernible effect on the already high immunoreaction found using any of the three antisera. It is concluded that the peptidase activity of acetylcholinesterase has the capacity to hydrolyze proteins of which some may be the precursor molecules for the enkephalins and substance P. Since the amacrine cells that contain the enkephalin-like and the substance P-like immunoreactivity were found to contain acetylcholinesterase, it is possible that the action found here in vitro represents a physiological function of the enzyme. The immunoreactivity on which there was no effect, somatostatin, does not co-exist with acetylcholinesterase. A second conclusion that may be drawn from these data is that the dark-adapted retinae lose immunoreactive peptide because of the rate of processing; the results suggest that there is adequate precursor molecule available to maintain "control" levels.     

Localization of vasoactive intestinal peptide immunoreactivity in human foetus and newborn infant spinal cord

Using an indirect immunofluorescence method the distribution of vasoactive intestinal peptide (VIP) immunoreactivity was studied in human foetus and newborn infant spinal cord and dorsal root ganglia. Further, for comparison some newborn infant brains were also investigated. Vasoactive intestinal peptide-like immunoreactive fibres were exclusively found in the caudal spinal cord and corresponding dorsal root ganglia. No immunoreactive cell bodies were detected. The first appearance of VIP-like immunoreactive fibres in both spinal cord and dorsal root ganglia was suggested during the fourth month of foetal life. Most immunolabelled fibres, concentrated in the sacral segment, were distributed in the Lissauer tract, along the dorsolateral gray border, in the intermediolateral areas and near the central canal in the dorsolateral commissure. A few VIP-like immunoreactive fibres were also seen in the dorsal funiculus and occasionally in the ventral gray horn and ventral roots. Further, a large population of VIP-like immunoreactive fibres occurs longitudinally in dorsal root, in ganglia and in the spinal nerve exit zone. These findings indicate the early appearance of VIP-like immunoreactive fibres in the human foetus spinal cord and corresponding ganglia. Moreover, they emphasize that in both foetus and newborn infant spinal cord VIP-like immunoreactive fibre distribution is limited to the lumbosacral segment.     

Distinctive structural and cytoskeletal properties of the long-surviving neurons in cell cultures of embryonic spinal cord

A distinctive population of neurons survives for longer than 3 months in cell cultures of chick or rat spinal cord. These neurons form a minor proportion (1%) of the neurons initially developing in the cultures, but are the only ones to survive longer than 30 days in vitro. In addition to their longevity, they share important morphological and cytoskeletal characteristics, which render them distinctive as a group even in early cultures which contain numerous other neurons of short-term viability. Each long-surviving neuron projects one neurite of great length relative to its other neurites, or to those of the shorter-lived neurons, and the length of this neurite is maintained constant for many weeks in vitro. This well-defined morphological feature may indicate the lineage(s) of these neurons. Structurally these cells are very different to the shorter-lived neurons. They are rich in neurofilaments and contain very few microtubules, whereas the shorter-lived neurons contain few neurofilaments but many microtubules. These differences in cytoskeleton coincide exactly with the distinction between limited and prolonged survival in vitro, and the possibility is considered that cytoskeletal stability in the presence of numerous small inflows of calcium might underlie "hardiness" in vitro. The state of development of the long-lived neurons is considered in the context of their shared features, and it is suggested that they may provide a model in which the regulation of development of neuronal function can be analysed.