An immunohistochemical localization of neuropeptide Y (NPY) in its amidated form in human frontal cortex

The distribution of neuropeptide Y (NPY)-immunoreactive neurons was studied in human frontal cerebral cortex from surgical biopsy specimens by immunohistochemical techniques. NPY-containing neurons were identified in all cortical sublayers except sublayer I. The stained neurons were of the multipolar, bitufted, round or triangular form with dendritic and axonal processes. The immunoreactive neurons were considered to be cortical interneurons, due to their nonpyramidal form, and since their processes could be followed intracortically particularly in direction to superficial cortical layers. The NPY precursor molecule is processed to NPY by a dibasic cleavage, and NPY is further enzymatically amidated before release and receptor activation can be achieved. Antisera raised against Cys-NPY(32-36)amide recognize amidated NPY not cross-reacting with nonamidated NPY. These antisera and immunohistochemistry revealed the presence of a population of NPYamide-immunoreactive cells morphologically indistinguishable from the NPY-immunoreactive cells in the human frontal cortex. By comparing the number of immunoreactive cells in adjacent sections, it appears that the number of NPY-immunoreactive cells was higher than those immunoreactive to NPYamide. Also, the density of NPY fibers was much higher compared with the number stained with NPYamide antiserum. The present immunohistochemical study indicates that NPY in its amidated form is contained in a subpopulation of human cortical NPY-immunoreactive neurons and may participate as an active neurotransmitter/modulator within the human cerebral cortex.     

Reaction of sensory neurons of the spinal ganglia to sectioning of their peripheral and central processes

Transversal ++semi-sectioning of the spinal ganglion (SG) is a good model for studying the reaction of the ganglional sensory neurons to sectioning of their peripheral and central processes. At sectioning the peripheral and central processes of the SG neurons degeneration of the neurons and their death take place. The degenerative processes are more pronounced in the neurons with the peripheral processes sectioned, and the restorative ones-with the central processes sectioned. The dynamics of the posttraumatic changes in absolute number of the neurons, profile areas of the body fields and neuronal nuclei, amount of neurons with certain signs of axonal reactions in the SG demonstrate a maximally pronounced reaction on the 7th day and beginning of restorative processes on the 15th day. They are not completed by the 180th day.     

The maturation of the somatostatin systems in the rat visual cortex

The development of somatostatin-immunoreactive (SS) neurons and processes in the rat visual cortex (VC) was studied in animals from embryonic day 20 (E20) to postnatal day 21 (D21). Three distinct patterns of immunoreactivity were seen. From E20 to birth (D0), VC was characterized by a small number of mainly bipolar SS neurons throughout the cortical plate. In the perinatal period, from D1 to D6, there were large numbers of immature immunoreactive neurons which were confined to layer VI and the subplate zone, a few bipolar neurons in the cortical plate and an extremely dense plexus of SS processes throughout the neuropil. The third phase, from D8 to weaning, was characterized by the absence of immature SS neurons, an increase in the number of multipolar SS neurons and a decrease in the density of SS fibers. By D15, the time of eye-opening, the number and distribution of SS neurons and processes was close to that seen in the adult. These results indicate that the SS system of neurons and fibers is among the earliest of the transmitter systems to be established in VC and suggests a role for the peptide in cortical organization as well as visual processing.     

Difference of pericentral NADPH-d positive neurons in the rabbit spinal cord segments

The purpose of the present investigation was to characterize and determine the number of NADPH-diaphorase positive neurons around the central canal in the rabbit spinal cord. These neurons are known to function as interneurons and are present in all spinal cord segments. They differ in shape of their bodies and in length and branching of their processes. The main differentiation was observed in their number, depending on the place of their localization. The highest number of these NADPH-diaphorase positive neurons was in sacral part (6 in average), the lowest one was noticeable in thoracic spinal cord (1-2 in average). It can be concluded that pericentral neurons of the rabbit spinal cord are capable of synthesizing nitric oxide and that they differ in number, depending on the place of their localization in each spinal cord segment.     

Early ontogeny of the vagus nerve: an analysis of the medulla oblongata and cervical spinal cord of the postnatal rat.

Retrograde transport of cholera toxin conjugated with horseradish peroxidase in the postnatal rat has revealed remarkable features of dendritic fields of vagal motor neurons in the medulla oblongata and cervical spinal cord during the period of early development (0-10 days). At birth, vagal motor neurons in the dorsal motor nucleus of the vagus, nucleus ambiguus, nucleus retroambigualis, nucleus dorsomedials and the spinal nucleus of the accessory nerve are small with relatively few, unbranched processes. The span of the dendritic tree is much smaller than that found in adult animals. By the postnatal Day 2 there are marked changes in the soma as well as in the dendritic tree of these neurons. There is dispersion of the cell bodies within the neuropil as well as an expansion of the total area of the brain stem occupied by these motor neurons and their dendritic processes which show extensive growth and branching. By postnatal Day 3 the most extensive proliferation of these neurons is seen and appears to represent the peak of dendritic growth of vagal motor neurons such that the area occupied by the dendritic tree of a single neuron is three times that seen in an adult rat. This proliferation gradually decreased during the subsequent seven days of early development (i.e. Days 4-10) so that by Day 10 the dendritic span of vagal motor neurons was reduced to about twice the adult size. This growth progressively decreased from Days 10 to 30 at which time adult levels were reached. Ultrastructural examination of these horseradish peroxidase labeled dendrites showed a positive correlation between the number of dendritic processes and the number of axo-dendritic synapses. This was accompanied by an increase in the number of identifiable synaptic junctions. These morphological complexities observed during the period of early development of vagal motor neurons indicate that the vagus nerve undergoes dramatic changes during the period of early development including the establishment of numerous synaptic contacts between vagal afferents and efferents in the brainstem. A number of these changes occur in developing dendritic fields of vagal motor neurons during the first three days of neonatal life. It is reasonable to assume that developmental abnormalities during this "critical period" could produce significant functional changes in the pattern of respiration as well as in the control of airway smooth muscle.     

The neural apparatus of the small intestine of piglets after administration of a growth stimulator kormogrisein]

By means of classical neurohistological techniques, phase contrast microscopy and morphometry, a comparative investigation has been performed concerning the development of the intramural nervous apparatus in the small intestine, normal and at application of cormogrisine. The structural peculiarities of morphogenesis are considered together with signs of activation and inhibition of the neurons growth in tissue culture. A number of morphological criteria, demonstrating an increased extrusive activity and enhancing potensity of the neurons growth have been revealed. The number of nervous processes becomes greater; degree of their ramification increases; a part of neurons of Dogiel II type turns into multiprocessive neurons with some signs of Dogiel I type cells; growth cones and arcadian structures are present; giant processes appear; thick nervous fasciculi are formed; volume of the neuron bodies increases more intensively. After application of cormogrisine for 2 months a definite neurostimulatory effect is revealed; it demonstrates a more intensive morphogenesis of the small intestine nervous of Physiology, USSR, Academy of Medical Sciences, Leningrad.     

人胎儿下丘脑结节区神经核团内生长抑素神经元的发生──免疫细胞化学研究

本实验收集２２例９周至２８周龄人胎儿,用免疫组织化学ＰＡＰ法研究了下丘脑结节区神经核团内生长抑素神经元的个体发生。结果表明。生长抑素免疫反应阳性神经元最早见于１６周人胚的弓状校及腹内侧核,以后随胎龄增长数目逐渐增加。在２２周时,腹内侧核内生长抑素免疫反应阳性神经元的数目达到高峰,弓状核内的该神经元在２４周达到高峰。２４周后阳性神经元数目呈递减趋向,免疫反应逐渐减弱。背内侧核及结节核内未见生长抑素免疫反应阳性神经元。生长抑素免疫反应阳性神经元形态多样,体积较小,其突起数目在发育过程中有较大变化。     

Modeling stability in neuron and network function: the role of activity in homeostasis

Individual neurons display characteristic firing patterns determined by the number and kind of ion channels in their membranes. We describe experimental and computational studies that suggest that neurons use activity sensors to regulate the number and kind of ion channels and receptors in their membrane to maintain a stable pattern of activity and to compensate for ongoing processes of degradation, synthesis and insertion of ion channels and receptors. We show that similar neuronal and network outputs can be produced by a number of different combinations of ion channels and synapse strengths. This suggests that individual neurons of the same class may each have found an acceptable solution to a genetically determined pattern of activity, and that networks of neurons in different animals may produce similar output patterns by somewhat variable underlying mechanisms.     

Influence of factors released from sciatic nerve on adult dorsal root ganglion neurons

Previous experiments have shown that medium conditioned (CM) by denervated peripheral nerve contains a process outgrowth promoting factor (s) for cultured adult frog dorsal root ganglion (DRG) neurons. The present experiments further characterize the influences of these factors on DRG neurons. The growth factors increases average process length by threefold, restricts the number of processes extended from four to two while simultaneously altering the morphology of those processes. Neurons with preexisting processes respond to the factors by significantly increasing the length of 35% of these processes. Only the newly elongated portions of preexisting processes have morphology typical of factor-induced processes, while the previously extended portions retain their original morphology. The number of processes of these neurons remains unchanged. Although composed of two population according to size, neurons in both populations are similarly influenced, suggesting that the factors influence neurons of all sensory modalities. To look at other possible influences of the nerve-released factors, a novel simple culture system has been developed in which concentration gradients of these factors can be established and maintained. The front of the outgrowth-promoting influence in these cultures could be followed over time (up to 9 days) as it affected the process length and morphology of neurons at increasing distances (up to 8 mm) from the source of the factors. The trophic factors may play important roles during regeneration in vivo by influencing the cytoskeletal organization in the cell body and growth cones to bring about a stabilization and consolidation of growth cone membrane of only a limited number of processes resulting in increasing the rate of process elongation. The factors may also serve to direct process outgrowth, which can be examined using the new culture system. 1994 John Wiley & Sons, Inc.     

Primary culture of lobster (Homarus americanus) olfactory sensory neurons

Lobster olfactory sensory neurons have contributed to a number of advances in our understanding of olfactory physiology. To facilitate further study of their function, we have developed conditions allowing primary culture of the olfactory sensory neurons in a defined medium. The most common cells in the culture were round cell bodies with diameters of 10-15 micro m that often extended fine processes, features resembling olfactory sensory neurons. We discovered that acetylcholinesterase acted as a growth factor for these cells, improving their survival in culture. We also confirmed previous evidence from spiny lobsters that poly-D-lysine was a superior substrate for olfactory cells of this size and morphology. We then identified olfactory sensory neurons in the culture in two ways. Almost half the cells tested responded to application of a complex odorant with an inward current. An even more rigorous test was made possible by the development of an antiserum to OET-07, an ionotropic glutamate receptor homolog specifically expressed by Homarus americanus olfactory sensory neurons. It labeled a majority of the round cells in the culture, unequivocally identifying them as olfactory sensory neurons.     

Distribution of tyrosine-hydroxylase-immunoreactive and dopamine-immunoreactive neurons in the central nervous system of the snail Helix pomatia

The distribution and characterization of dopamine-containing neurons are described in the different ganglia of the central nervous system of Helix on the basis of the distribution of tyrosine hydroxylase immunoreactive (TH-ir) and dopamine immunoreactive (DA-ir) neurons. Both TH-ir and DA-ir cell bodies of small diameter (10–25 m) can be observed in the buccal, cerebral and pedal ganglia, dominantly on their ventral surface, and concentrated in small groups close to the origin of the peripheral nerves. The viscero-parietal-pleural ganglion complex is free of immunoreactive cell bodies but contains a dense fiber system. The largest number of TH-ir and DA-ir neurons can be detected in the pedal, and cerebral ganglia. The average number of TH-ir and DA-ir neurons significantly differs but all the identifiable groups of TH-ir neurons also show DA-immunoreactivity. Therefore, we consider the TH-ir neurons in those groups as being DA-containing neurons. The amounts of DA in the different ganglia assayed by high performance liquid chromatography correspond to the distribution and number of TH-ir and DA-ir neurons in the different ganglia. The axon processes of the labeled small-diameter neurons send thin proximal branches toward the cell body layer but only rarely surround cell bodics, whereas distally they give off numerous branches in the neuropil and then leave the ganglion through the peripheral nerves. In the cerebral ganglia, the analysis of the TH-ir pathways indicates that the largest groups of labeled neurons send their processes through the peripheral nerves in a topographic order. These results furnish morphological evidence that DA-containing neurons of Helix pomatia have both central and peripheral roles in neuronal regulation.     

Age-related changes of parvalbumin immunoreactive neurons in the rat main olfactory bulb

Parvalbumin (PV) is found in the olfactory system, including the main olfactory bulb, and is thought to be one of the neuroactive substances in olfaction. Changes in PV immunoreactivity in the olfactory system during aging have not been examined. We investigated such changes in the main olfactory bulb (MOB) of the rat at postnatal month 1 (PM 1), PM 3, PM 6, PM 12 and PM 24. PV-IR neurons were almost completely restricted to the external plexiform layer. At PM 1 there were only a few PV-IR neurons; at PM 3, the number of PV-IR neurons was at its greatest but they were not well developed morphologically. At PM 6, the number of PV-IR neurons was similar to that at PM 3 and they had satellite somata with well-developed processes with many varicosities. By PM 12 the number of neurons and processes had declined, and by PM 24, they had fallen even further and the remaining processes had lost most of their varicosities. We conclude that age-related degeneration of PV-IR neurons in the MOB may reduce calcium buffering and affect olfactory function in senile species.     

Structural organization of procerebrums of terrestrial molluscs: Characteristics of neuronal pattern, plasticity, and age peculiarities

In terrestrial snails and slugs, the presence of five types of neurons has been shown: typical granular, horizontal, stellate, apical, and basal cells. A peculiarity of procerebrum neurons is a loop-like segment of the basal process. Granular cells have been established to spread both as cell columns and as single cells. The number of the columns and the number of cells in each column depend on the age and individual peculiarities of the animal. In the course of ontogenesis, there also occur changes of the number of processes-outgrowths of granular cell bodies as well as of the character of thickenings and the type of endings on their main processes. Basal neurons and single granular cells of the columns form several independent tracts running from the procerebrum cell body areas to metacerebrums. The single granular cells, horizontal cells, and a part of basal neurons are considered as associative elements providing formation in procerebrums of the single system that is a basis for the synchronized activity of the whole neuronal complex. A similarity of organization of procerebrums and of associative-integrative centers of higher animals is discussed.     

Remodeling of astrocytes, a prerequisite for synapse turnover in the adult brain? Insights from the oxytocin system of the hypothalamus

Neurons, including their synapses, are generally ensheathed by fine processes of astrocytes, but this glial coverage can be altered under different physiological conditions that modify neuronal activity. Changes in synaptic connectivity accompany astrocytic transformations so that an increased number of synapses are associated with reduced astrocytic coverage of postsynaptic elements, whereas synaptic numbers are reduced on reestablishment of glial coverage. A system that exemplifies activity-dependent structural synaptic plasticity in the adult brain is the hypothalamo-neurohypophysial system, and in particular, its oxytocin component. Under strong, prolonged activation (parturition, lactation, chronic dehydration), extensive portions of somatic and dendritic surfaces of magnocellular oxytocin neurons are freed of intervening astrocytic processes and become directly juxtaposed. Concurrently, they are contacted by an increased number of inhibitory and excitatory synapses. Once stimulation is over, astrocytic processes again cover oxytocinergic surfaces and synaptic numbers return to baseline levels. Such observations indicate that glial ensheathment of neurons is of consequence to neuronal function, not only directly, for example by modifying synaptic transmission, but indirectly as well, by preparing neuronal surfaces for synapse turnover.     

A Neurosecretory System in the Brain of the Lancelet, Branchiostoma lanceolatum

Ultrastructural and histochemical studies indicate a neurosecretory system exists in the lancelet brain with basal properties resembling a primitive hypothalamic system. A nucleus of secreting neurons, containing peptide granules (115 nm), is prominent in the dorsal walls of the brain. The axons establish contacts with the ventral brain surface, probably releasing their secretory product out of the brain. The neurons are innervated by dopaminergic "boutons en passant" often very active with a high number of electron translucent vesicles as well as dense-core vesicles (90 nm). Ventrally located cellbodies containing what are probably secretory peptide granules (110 nm) establish contacts with their basal processes on the ventral brain surface.     

A light and electron microscopic study of the preoptic nucleus of the grass frog (Rana pipiens), under normal conditions and following transection of the preoptico-neurohypophysial tract

Degenerative and regenerative processes occur in the preoptic neurons following transection of the preoptico-neurohypophysial tract. Three types of responses after transection were observed: affected, recovered, and degenerated neurons. However, transection of the tract did not stop the synthesis of neurosecretory granulated vesicles. The affected neurosecretory neurons showed nuclear changes, increased number of Golgi complexes, and dilated cisternae of rER, as well as, an increased number of dense bodies. The recovered neurosecretory neurons contained long non-dilated cisternae of rER which were organized in a concentric manner. Also seen were large nuclei with evenly distributed chromatin, less active Golgi complexes, and vesicles. The degenerated neurosecretory neurons exhibit pyknotic nuclei, a net of dilated cisternae of rER, dense bodies, and electron dense cytoplasm.     

Cell shape and motility of oligodendrocytes cultured without neurons

Summary Oligodendrocytes, the myelin-forming cells of the central nervous system (CNS), were cultured from newborn rat brain and optic nerve to study how they differentiate in vitro in the absence of neurons. By use of galactocerebroside (GC) as a reference marker, the development of the cell phenotype was studied with video-enhanced differential interference contrast microscopy, immunofluorescence and electron microscopy. After a few days in culture, oligodendrocytes extend 5 to 10 main processes that are very rich in microtubules, but they did not stain with a monoclonal antibody reacting with all known classes of intermediate filaments. The number of processes can vary with the substrate on which the cells are grown; fewer processes form on laminin than on polylysine coated glass. Oligodendrocytes, in a fashion similar to that of neurons appear to keep their body immobile while the long processes grow. However, while neurons display motile activities mostly at the end of the cell processes called growth cones, the oligodendrocytes display motile, actin rich filopodia and lamellipodia along the entire length of all processes. The outgrowth of motile processes from oligodendrocytes sometimes occurs preferentially towards neighboring astrocytes. Oligodendrocyte processes display intense bidirectional movement of cytoplasmic organelles. Movement of surface components also occurs since GC molecules cross-linked by antibodies move from the processes towards the cell body. Thus, oligodendrocytes cultured without neurons develop on schedule a complex phenotype similar to their in vivo counterpart. In addition, their processes are capable of specific motile activities which may function in vivo to find the target axon and to transport myelin membrane components at the site of myelin assembly.Abbreviations (CNS) Central nervous system - (DIC) Differential interference contrast - (GC) Galactocerebroside - (GFA) protein Glial fibrillary acidic - (NSE) Neuron-specific enolase     

Relative number and distribution of murine hypothalamic proopiomelanocortin neurons innervating distinct target sites

Proopiomelanocortin (POMC) neurons send projections widely throughout the brain consistent with their role in regulating numerous homeostatic processes and mediating analgesia and reward. Recent data suggest that POMC neurons located in the rostral and caudal extents of the arcuate nucleus of the hypothalamus may mediate selective actions, however it is not clear if POMC neurons in these regions of the arcuate nucleus innervate specific target sites. In the present study, fluorescent microspheres and cholera toxin B were used to retrogradely label POMC neurons in POMC-DsRed transgenic mice. The number and location of POMC cells projecting to the supraoptic nucleus, periaqueductal gray, ventral tegmental area, paraventricular nucleus, lateral hypothalamic nucleus, amygdala and the dosal vagal complex was determined. Tracer injected unilaterally labeled POMC neurons in both sides of the arcuate nucleus. While the total number of retrogradely labeled cells in the arcuate nucleus varied by injection site, less than 10% of POMC neurons were labeled with tracer injected into any target area. Limited target sites appear to be preferentially innervated by POMC neurons that reside in the rostral or caudal extremes of the arcuate nucleus, whereas the majority of target sites are innervated by diffusely distributed POMC neurons. The modest number of cells projecting to each target site indicates that relatively few POMC neurons may mediate potent and specific physiologic responses and therefore disturbed signaling in a very few POMC neurons may have significant consequences.     

Differences in the cyto- and angioarchitectonics of the ventral horns of the spinal cord in higher and lower vertebrates

When comparing structures of the ventral horns of the spinal cord in amphibia (Rana esculenta) and Carnivora (cat, dog), it has been stated that in the latter, together with increasing number of neurons their blood supply is improving. This is demonstrated as a growing density of the capillary network in the cerebral substance and increasing number of capillaries within 25 mcm around the neuronal bodies. Glial surrounding of the neurons changes considerably. Both in the amphibia and Carnivora astrocytes and oligodendrocytes play the role of neuronal satellites. But in the amphibia astrocytes as satellites occur much more often than oligodendrocytes. This is evidently connected with a low blood supply and with certain peculiarities of metabolic processes in the cerebral tissue.