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Development of corticospinal tract fibers and their plasticity I: quantitative analysis of the developing corticospinal tract in mice.

This study was undertaken to elucidate ultrastructurally and quantitatively the development of the corticospinal tract (CST) axons of mouse at the intumescence level of the cervical cord. An anterograde HRP study showed that the CST was located at the ventral one-third of the dorsal funiculus, and a few HRP-positive fibers were noted at the medialmost part of the ipsilateral anterior funiculus. Ultrastructurally, the CST was composed of unmyelinated axons, growth cones and a few degenerating axons until postnatal day 10 (P10), then the axons in CST gradually increased in size. The number of axons constituting the right CST was calculated at different days of age. The total numbers of axons at P0, P4, P14, P21 and P56 were 2.3 x 10(4), 6.2 x 10(4), 10.4 x 10(4), 7.1 x 10(4) and 3.5 x 10(4), respectively. These results indicate that the number of CST axons at the cervical intumescence of mouse becomes maximum at P14, and then decreases rapidly to reach the adult level of 3.5 x 10(4) (at P56), about 68% of them thus being lost.

Animals↗

Inhibition of nitric oxide synthase fails to disrupt the development of cholinergic fiber patches in the rat superior colliculus.

Nitric oxide may serve as a retrograde messenger to refine or stabilize synapses in the developing nervous system. Whether this action is dependent upon glutamate and the N-methyl-D-aspartate receptor is not yet established. We have used the patch-cluster system in the intermediate gray layer (IGL) of the rat superior colliculus (SC), a system receiving both glutamatergic and cholinergic input, to study this question. The normal distribution and development of nitric oxide synthase (NOS) in SC was examined using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry in Sprague-Dawley rats aged P4 to adulthood. Fibers containing acetylcholine (ACh) were identified using choline acetyltransferase (ChAT) immunocytochemistry. In addition, N omega-nitro-L-arginine, an inhibitor of NOS, was injected intraperitoneally from birth until P10, P14, P18, or P21-22 to determine if NOS inhibition would disrupt the formation of the ACh patches. Control animals were studied from the same age groups. Our results show NADPH-d-labeled cells within the periaqueductal gray and the deep gray layer of SC by P4, the earliest age examined. By P8-P9, cells in the IGL were well labeled by NADPH-d, while few in the superficial layers (SL) were labeled. SL cells were visible by P10 and were intensely labeled by P14. IGL cells transiently expressed NADPH-d in that the number of labeled cells increased from P8 to P35, then decreased in the adult. ChAT-labeled fibers first appeared in the IGL at P10, formed a characteristic two-tier pattern by P14, and established obvious patches by P21. Inhibition of NOS from birth produced no qualitative differences in the distribution or density of either ChAT-labeled fibers or NADPH-d-labeled cells and fibers at any of the ages examined. We therefore conclude that NO does not contribute to the refinement of cholinergic fiber patches in the rat SC, probably because the fiber system is not glutamatergic.

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Extensive target cell loss during development results in mossy fibers in the regio superior (CA1) of the rat hippocampal formation.

The axons of dentate granule cells (mossy fibers) have been reported to appear in the regio superior (CA1) of the rat hippocampal formation following destruction of the pyramidal cells in the regio inferior (CA3). We undertook the present experiments to confirm this finding and to determine the requirements for this dramatic neuronal rearrangement. We found that extensive (greater than 80%) loss of CA3 cells, as well as the presence of surviving CA1 neurons within a narrow period of development (postnatal days 3-5) is necessary, however apparently not sufficient, for the appearance of CA1 mossy fibers. That the absence of normal target cells during a restricted period of mossy fiber development will lead to their association with novel targets suggests that much of the specificity of this developing connection depends on the presence of normal targets during a critical period.

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Histochemical evidence of altered development of cholinergic fibers in the rat dentate gyrus following lesions. I. Time course after complete unilateral entorhinal lesion at various ages.

The entorhinal cortex of rats was removed at various times during development, and the reaction of the cholinergic septohippocampal input to the dentate gyrus was examined by use of acetylcholinesterase histochemistry. When the ipsilateral entorhinal cortex is completely removed, the outer 70-75% of the molecular layer of the dentate gyrus is almost completely denervated. After such a lesion at 5 to 33 days of age, the acetylcholinesterase staining initially intensified throughout the denervated area, indicating that the septohippocampal fibers branched or elongated. This reaction could be detected within one day after a lesion at 11 days of age and within three or five days after lesions at earlier or later times. Whereas the initial response of the septohippocampal fibers was independent of the age at which the lesion was made, their final localization depended on the developmental state of the animal. After lesions at the age of 5 or 11 days, the reactive septohippocampal fibers became restricted to the outer one-sixth to one-third of the molecular layer within two days after appearance of their initial reaction. A similar concentration of reactive fibers was demonstrable after lesions at 16, 18 or 21 days of age, but some reaction persisted in the middle third of the molecular layer. Finally, after lesions at 26 or 33 days of age the proliferating cholinergic fibers ultimately were uniformly distributed throughout the outer 60% of the molecular layer. These results suggest that septohippocampal fibers initially extend or sprout throughout the denervated area to replace the lost perforant path fibers. However, the reactive fiber population becomes restricted to the outer edge of the molecular layer if the entorhinal lesion is made before the period of cholinergic synaptogenesis and concentrates in this same zone if it is made while cholinergic synapses are forming. We suggest that either the proliferative reaction continues in the outer part of the molecular layer and subsides in other parts of the denervated area or septohippocampal fibers move outward through the molecular layer to assume a more superficial location. After entorhinal lesions at 16 days of age or later the pale-staining zone (containing fibers that originate in hippocampus regio inferior) immediately deep to the denervated area widened. If the lesion was made earlier, this zone never developed at most septotemporal levels of the dentate gyrus. These results are probably related to the extension of regio inferior fibers into the denervated area.

Acetylcholinesterase↗

Effect of ATP concentration and pH on rigor tension development and dissociation of rigor complex in glycerinated rabbit psoas muscle fiber.

Isometric rigor tension development of glycerinated rabbit psoas muscle fibers in a medium, due to the formation of rigor complexes, was estimated at varying ATP concentrations from 0 to 2.5 mM and pH values from 6.75 to 8.20. The dissociation of rigor complexes was also estimated under the same conditions. When muscle fibers developed rigor tension from the relaxed and rigor states, the magnitude of rigor tension increased with increasing concentration of ATP. Transition between rigor and relaxation in single fibers occurred discontinuously at constant levels (critical levels) of ATP which were determined by pH. The critical concentrations of ATP necessary for inducing the transitions between rigor and relaxed states were also increased exponentially with increased pH. Incomplete repetition of tension development by the same fiber was also observed. This incomplete reversibility was divided into two types: one which showed a decay in rigor tension and another which showed no decay. The reason for the incomplete reversibility was discussed

Adenosine Triphosphate↗

Transient expression of a ventricular myosin heavy chain isoform in developing chicken intrafusal muscle fibers.

Sections of chicken tibialis anterior and extensor digitorium longus muscles were incubated with monoclonal antibodies against myosin heavy chains (MHC). Ventricular myosin was present in developing secondary intrafusal myotubes when they were first recognized at embryonic days (E) 13-14, and in developing extrafusal fibers prior to that date. The reaction in intrafusal fibers began to fade at E17, and in 2-week-old postnatal and older muscles the isoform was no longer recognized. Only those intrafusal fibers which also reacted with a monoclonal antibody against atrial and slow myosin contained ventricular MHC. Intrafusal myotubes which developed into fast fibers did not express the isoform. Hence, based on the presence or absence of ventricular MHC, two lineages of intrafusal fiber are evident early in development. Strong immunostaining for ventricular MHC was observed in primary extrafusal myotubes at E10, but the isoform was already downregulated at E14, when secondary intrafusal myotubes were still forming and expressed ventricular MHC. Only light to moderate and transient immunostaining was observed in coexisting secondary extrafusal myotubes, most of which developed into fast fibers. Thus at the time when nascent muscle spindles are first recognized, differences in MHC profiles already exist between prospective intrafusal and extrafusal fibers. If intrafusal fibers stem from a pool of primordial muscle cells, which is common to intrafusal and extrafusal myotubes, they diverged from it some time prior to E13.

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Fiber and the development of gastrointestinal function.

Dietary lipid represents a major calorie source for the developing infant, yet dietary lipid absorption is relatively inefficient by adult standard. Recent studies are presented, indicating that the developmentally related improvement in lipid absorption observed in the preterm and fullterm infant relate to the maintenance of an intact intraluminal phase and completion of the enterohepatic circulation for bile salts. Critical issues including the absorption of nonpolar vitamins, iron, and the type of dietary lipid are discussed. The possible influence of the introduction of dietary fiber to the food of the developing infant is examined, and areas for future investigation are discussed.

Cellulose↗

ATP-induced tension development in glycerinated fibers of scallop adductor striated muscle. Role of regulatory light chain of myosin in calcium regulation of muscle contraction.

Removal of the regulatory light chain subunit (EDTA light chain) of myosin from glycerinated fibers of scallop adductor striated muscle resulted in an immediate loss of calcium sensitivity of tension development and in a subsequent decrease in tension developed in the presence of calcium. It is suggested that removal of EDTA light chain results in a change in the myosin heavy-chain conformation which is probably responsible for the decrease in tension development.

Adenosine Triphosphate↗

Prenatal development of rat primary afferent fibers: I. Peripheral projections.

Development of the peripheral innervation patterns of the L1-S1 lumbosacral ganglia and motor segments in embryonic day 12-17 (E12-17) rat embryos was examined using carbocyanine dyes. Individual dorsal root ganglia (DRGs) and/or isolated ventral horn (VH) segments, or individual peripheral nerves, were isolated in rat embryos fixed at different stages and filled with one of three carbocyanine dyes; DiI, DiA, and DiO. Individual experimental preparations included labeling of 1) single DRGs; 2) multiple DRGs with alternating dyes, DiO, DiI, and DiA; 3) single isolated VH segments; 4) multiple VH segments with alternating dyes; 5) single VH segments and the corresponding segmental DRGs with different dyes; and 6) two or more individual peripheral nerves labeled with different dyes. Results from these preparations have shown that the first fibers exited the lumbar ventral horn and DRGs at E12. At E13 major nerve trunks (e.g., femoral and sciatic) were visible as they exited the plexus region. By E14 afferent fibers were present in the epidermis of the proximal hindlimb, and the major nerve trunks extended into the leg. Fibers originating from L3 to L5 (DRG and VH) reached the paw by E14.5-E15, and the epidermis of the most distal toes was innervated by E16-E16.5. While afferent fibers and motor axons of the same segmental origin mixed extensively in the spinal nerve, fibers of different segmental origin combined in the plexus and major nerve trunks with little or no interfascicular mixing. Dermatomes observed at E14 were in general spotty and non-contiguous. However, by E16 the dermatomes resembled mature forms with substantial overlap only between adjacent ones. Thus the adult pattern of spatial relationships between cutaneous afferent fibers in the periphery is established early in development.

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Both basic fibroblast growth factor and ciliary neurotrophic factor promote the retention of polyneuronal innervation of developing skeletal muscle fibers.

At birth, nearly all rat muscle fibers receive synaptic inputs from more than one motoneuron at a single end-plate site. By the end of the third postnatal week all but one of these inputs has been eliminated. During this loss of polyneuronal innervation, developing neuromuscular synapses compete with one another. Although the nature of this competition is not known, it is commonly assumed that it is mediated through differential activity of the competing inputs. One means by which such differential activity might be translated into a biological signal would be if the synapses compete in an activity-dependent manner for a scarce supply of neurotrophic molecules. A prediction of this hypothesis is that excess quantities of such trophic molecules will reduce competition and thereby slow down or abolish the normal loss of polyneuronal innervation. In newborn rats, the effects of injection of either basic fibroblast growth factor (bFGF) or ciliary neurotrophic factor (CNTF) on the outcome of neuromuscular synapse elimination were investigated. Daily injections of either bFGF or CNTF were made for 1 week into the lateral gastrocnemius muscle beginning at the postnatal age of 2 days. The amount of polyneuronal innervation of fibers in trophic molecule-injected muscles and saline-injected contralateral muscles was assayed using intracellular recording methods. For both bFGF- and CNTF-injected muscles, an increase in the percentage of polyneuronally innervated fibers relative to saline-injected muscles was noted. For bFGF-injected muscles, the amount of polyneuronal innervation remained at nearly 60% as late as the postnatal age of 14 days (P14). This is the amount of polyneuronal innervation found at age 6 days in normal animals. Nearly 40% of the fibers of CNTF-injected muscles remained polyneuronally innervated at age P14, the amount expected at age 9 days. These results indicate that both bFGF and CNTF exert powerful and long-lasting effects on developing neuromuscular synapses.

Animals↗

Failure to disrupt development of cholinergic fiber patches in the superior colliculus in nitric oxide synthase deficient mice.

Nitric oxide (NO) has been shown to mediate refinement of glutamatergic axonal pathways during development. In this study, we investigated whether the development of a cholinergic pathway in the intermediate gray layer (IGL) of the mouse superior colliculus (SC) is also mediated by NO. The pathway was labeled using an antibody directed against choline acetyltransferase (ChAT) and its distribution examined in normal C57/BL6 mice and in knockout mice in which the genes for the neuronal isoform of nitric oxide synthase (NOS) or both the endothelial and neuronal isoforms of NOS had been disrupted. We also examined the development of expression of NOS using nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) staining. NADPHd labeled cells were found within the IGL by P8 and formed loose clusters of cells by P12-P15. ChAT and NADPHd labeled fibers were first observed at P12 and gradually established their characteristic two-tiered patchy pattern between P14 and P21. Comparison of the ChAT labeled fiber distribution in normal, single nNOS and double e,nNOS knockout mice revealed no differences between these three groups. We therefore conclude that nitric oxide does not mediate refinement of this cholinergic pathway.

Animals↗

Effect of highly fermentable dietary fiber on the development of swine dysentery and on pig performance in a "Pure--Culture Challenge Model".

This study tried to evaluate the effect of highly fermentable fiber on the incidence and severity of swine dysentery (SD) after experimental oral infection with pure cultures of Brachyspira (B.) hyodysenteriae. Forty eight growing pigs were allocated to two groups and treated until slaughter as follows: Group 1 (n = 24): infected with B. hyodysenteriae and fed with a food containing 9.6% highly fermentable neutral detergent fiber. Group 2 (n = 24): infected with B. hyodysenteriae and fed with a food containing 6.1% low fermentable neutral detergent fiber. Pigs of each group were intragastrically inoculated on each of three consecutive days with pure culture of 1.8 x 10(10) B. hyodysenteriae. All pigs were monitored daily until slaughter. Faecal shedding of B. hyodysenteriae by polymerase chain reaction, antibody response by IFA, clinical signs, growth performance and extents of gross and microscopical lesions specific for swine dysentery were determined. Faecal shedding of B. hyodysenteriae and antibodies specific for B. hyodysenteriae were detected at day 30 post infectionem. Significant (p < 0.05) milder clinical signs typical for swine dysentery were detected in group 1, fed with 9.6% high fermentable fiber compared to group two fed with a food containing 6.1% low fermentable neutral detergent fiber. Daily weight gain differed significantly (p < 0.05) between the groups (group one 780 g vs. group two 760 g). Food conversion efficiency showed in group one a significant (p < 0.05) better (3.28) result than in group two (3.38). Feed consumption presented significantly (p < 0.001) better results in group one compared to group two (2.38 kg vs. 2.25 kg). From our experimental findings we conclude that in production units suffering of swine dysentery high levels of highly fermentable fiber in diet may increase health and performance.

Animals↗

Comparison of rat and mouse pulmonary tissue mechanical properties and histology.

The present study compares the dynamic mechanical properties and the contents of collagen and elastic fibers (oxytalan + elaunin + fully developed elastic fibers) of mice and rat lung strips. Resistance, elastance (E), and hysteresivity (eta) were obtained during sinusoidal oscillations. The relative amounts of blood vessel, bronchial, and alveolar walls, as well as the mean alveolar diameter were determined. In both species, resistance had a negative and E a positive dependence on frequency, whereas eta remained unchanged. Mice showed higher E and lower eta than rats. Although collagen and elastic fiber contents were similar in both groups, mice had more oxytalan and less elaunin and fully developed elastic fibers than rats. Rats showed less alveolar and more blood vessel walls and higher mean alveolar diameter than mice. In conclusion, mice and rats present distinct tissue mechanical properties, which are accompanied by specific extracellular fiber composition.

Airway Resistance↗

The bright side of JNKs-Multitalented mediators in neuronal sprouting, brain development and nerve fiber regeneration.

The c-Jun N-terminal kinases (JNKs) are important regulators of physiological and pathological processes in the central and peripheral nervous system. In general, JNKs are considered as mediators of neuronal degeneration in response to stress and injury. However, recent data have provided substantial evidence that JNKs are also essential for physiological and regenerative signalling in neurons. This review summarizes the importance of JNKs for neurite formation and outgrowth, brain development, dendritic architecture and regeneration of nerve fibers after injury. We discuss putative mechanisms which control the bipartite actions of individual JNK isoforms for neuronal death and repair after nerve fiber injury with a particular focus on the role of the transcription factor c-Jun.

Animals↗

Nerve growth factor accelerates seizure development, enhances mossy fiber sprouting, and attenuates seizure-induced decreases in neuronal density in the kindling model of epilepsy.

Recurrent seizure activity induced during kindling has been reported to produce a functional synaptic reorganization of the mossy fibers in the hippocampus. To date, it is unclear whether this kindling-induced growth is secondary to decreases in hilar neuron density, which are presumed to reflect hilar neuronal cell loss, or whether it is related specifically to an activation-dependent plasticity. We recently demonstrated that blocking nerve growth factor (NGF) biological activity retards seizure development and inhibits the sprouting of mossy fibers. We now demonstrate that intraventricular administration of NGF itself accelerates the progression of kindling epileptogenesis, increases mossy fiber sprouting in the CA3 region and in the inner molecular layer (IML), but reduces seizure-induced decreases in hilar cell density. These findings provide support for a role of NGF in kindling and kindling-induced mossy fiber sprouting. In addition, the results dissociate this form of epileptogenesis from hilar cell loss or decreases in hilar cell density attributable to increases in hilar area, thereby supporting seizure-induced mossy fiber sprouting as being primarily attributable to the combined effects of neuronal activation and the activation-induced upregulation of growth factors.

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Variations in the tangential distribution of postsynaptic glutamate receptors in Purkinje cell parallel and climbing fiber synapses during development.

Major factors affecting the responses of a neuron to release of glutamate include the kinds and distribution of glutamate receptors in the neuron and their distribution along the surface of the postsynaptic membrane (tangential distribution). The latter distribution pattern is established during the development of the synapse and could be modified during maturation of synapse structure/function and through adult synapse plasticity. Parallel and climbing fiber synapses of cerebellar Purkinje cells are good models for studying this pattern because they contain two major kinds of ionotropic glutamate receptors, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) and delta, that are involved in adult plasticity and show differences in distribution, and because these two synapse types show complex changes in architecture and glutamate receptor distributions during development. In the present study, both AMPA and delta receptors showed variations in tangential distributions during many stages of development from postnatal day 2 to adult; i.e. qualitative assessment showed that receptors are concentrated either near the center or in outer portions of the synapse, while they are rare or absent from the perisynaptic region. Quantitative analysis showed statistically significant nonuniformities at some ages; the most common nonuniformity in these cases appears to be a drop-off in receptor density in the outer 20% of the synapse. Statistical analyses also indicated that distribution patterns did not change significantly with age. In contrast to the ionotropic receptors, the metabotropic glutamate receptor, mGluR1alpha, was found mainly in the perisynaptic region both during development and in adults. Differences in the distribution of glutamate receptors may be necessary to assure an effective response to glutamate release and may be modified through synaptic plasticity. Our findings show that the basic patterns of distribution of ionotropic and metabotropic glutamate receptors in synapses are established early in development, indicating that the postsynaptic density/membrane region is highly organized even in the immature synapse.

Animals↗