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Characterization of the postnatal development of superior laryngeal nerve fibers in the postnatal kitten.

A combined electron microscopic and electrophysiological study of the superior laryngeal nerve (SLN) was undertaken in postnatal kittens ranging in age from 1-63 days. The superior laryngeal nerve is predominantly a sensory nerve innervating the upper respiratory tract, and could play a potential role in the modulation of respiration, particularly in the infant animal. Distribution of fibers in the developing SLN indicates that within the first postnatal month, 75% of the fibers are unmyelinated, and by 42 days, the myelinated fibers increase in number to approximately 50%. Of the myelinated fibers present in the one day old kitten, 3-4% of those exceeded 4 mum in total diameter, which is the minimum diameter for normal conduction velocity of action potentials. The distribution of the diameter sizes of the myelinated fibers is bell-shaped within the first 45 days after which the curve becomes skewed to the right (43-61 days; mean 2.6 mum, range 0.5-8.0 mum) to resemble the adult distribution of myelinated fibers (mean 4.2 mum, range 1.6-13.0 mum). Two variable plots of myelin width to axon diameter suggest a steeper slope for developing fibers as compared to that of the adult fibers. Electrical stimulation of the sectioned SLN indicates that evoked potentials could be recorded from the recurrent laryngeal nerve innervating the laryngeal intrinsic muscles and from the hypoglossal nerve to the tongue musculature in the youngest kittens tested (i.e., age 9 days). Stimulation at selected frequencies of 3 and 30/sec readily evoked apnea in the youngest kitten studied (i.e., age 5 days), while swallowing was more readily evoked at 28-30 days when using electrical stimulation.

Age Factors↗

Expression of GFAP immunoreactivity during development of long fiber tracts in the rat CNS.

Astrocyte maturation in the developing corpus callosum and dorsal columns of the spinal cord was studied immunocytochemically in the rat, using antiserum to glial fibrillary acidic protein (GFAP) with a view to determining the relationships of astrocytes to the advancing axons of the corpus callosum and corticospinal tract. Between the eighteenth and nineteenth days of gestation, when the corpus callosum commences forming, most of the GFAP staining in the cerebral hemispheres is contained in radial processes, but some staining of glial cell bodies is also seen in the ventricular zone. At the region of interhemispheric fusion, where the corpus callosum will form, an accumulation of astrocytic processes demonstrable electron microscopically shows light immunocytochemical staining for GFAP. These processes do not adopt a stereotyped orientation. Rather, the overall impression as one moves towards the midline, is of radially disposed processes being disrupted and disoriented by the growing callosal axons at the fusion of the hemispheres. At no time can any orderly arrangement of GFAP-containing processes be seen which might indicate that the processes are serving to guide the growing axons across the midline. There is no immunoreactive staining of cell bodies or processes ventral to the corpus callosum, except in postnatal animals. Prior to the arrival of corticospinal axons in the spinal cord on the first postnatal day (PO)21, GFAP immunoreactivity is greatest in radial processes of the lateral funiculi and in the dorsal median septum. Oblique or vertical processes increase in the cuneate fasciculus from P0 tot P4 but do not appear in the gracile fasciculus until P4. Virtually no stained processes appear in the region to be traversed by the principal corticospinal tract, nor later in the tract itself until late in postnatal development. Only by 3 weeks postnatal is the adult pattern of GFAP staining observed in the corticospinal tract. These results also indicate that the expression of GFAP immunoreactivity is a relatively late phenomenon in astrocytes associated with advancing axons and implies that this aspect of astrocytic maturation is unrelated to any guidance that the immature astrocytes might provide for the growing axons.

Animals↗

Structure and chemical composition of bast fibers isolated from developing hemp stem.

Microscopic and chemical changes of hemp bast fibers were studied during the maturation from vegetative to grain maturity stages at both apical and basal regions of the stems. The content of protein was the main factor related to fiber maturation, whereas increased proportions of mannose and glucose and decreasing levels of galactose were also highly significant. Enhanced glucose deposition in apical fibers could be related to the gradual thickening of the fibers, whereas in basal regions the thickness of the fibers nearly reached the maximum at vegetative stages. In contrast, the extent of lignification remained close to 3-4% during plant growth. Hemp fiber lignins were rich in guaiacyl units and would be rather condensed in nature. In addition, the proportion of p-hydroxyphenyl units displayed a constant decline during maturation. A progressive chemical fractionation of hemp fibers provided further insights to the occurrence and nature of noncellulosic polysaccharides. Notably, these data pointed out that maturation is accompanied by a significant increase in water- and alkali-soluble components containing glucose- and mannose-related polymers and a decrease in arabinose and galactose components disrupted by diluted hydrochloric acid. Taken together, chemical features of the noncellulosic components suggest that the architecture of hemp fibers differs slightly from that of the more widely studied flax fibers.

Cannabis↗

Human skeletal muscle fiber types: delineation, development, and distribution.

This brief review attempts to summarize a number of studies on the delineation, development, and distribution of human skeletal muscle fiber types. A total of seven fiber types can be identified in human limb and trunk musculature based on the pH stability/lability of myofibrillar adenosine triphosphatase (mATPase). For most human muscles, mATPase-based fiber types correlate with the myosin heavy chain (MHC) content. Thus, each histochemically identified fiber has a specific MHC profile. Although this categorization is useful, it must be realized that muscle fibers are highly adaptable and that innumerable fiber type transients exist. Also, some muscles contain specific MHC isoforms and/or combinations that do not permit routine mATPase-based fiber typing. Although the major populations of fast and slow are, for the most part, established shortly after birth, subtle alterations take place throughout life. These changes appear to relate to alterations in activity and/or hormonal levels, and perhaps later in life, total fiber number. Because large variations in fiber type distribution can be found within a muscle and between individuals, interpretation of data gathered from human muscle is often difficult.

Adenosine Triphosphatases↗

Does axonal branching contribute to the overproduction of optic nerve fibers during early development of the cat's visual system?

The number of axons within the optic nerve of the fetal cat was found to be no greater near the chiasm than near the eye. This suggests that the overabundance of axons present in the developing nerve results from a corresponding excess of retinal ganglion cells, and that the elimination of optic fibers that occurs during normal development is principally due to the loss of ganglion cells and not to the elimination of axonal branches.

Animals↗

[The embryological development of the nerve fibers of the tooth. An analysis of their formation and development correlated with the different evolutionary stages of the dental structures].

In this review the research on the growth and development of nerve fibers in the dental pulp have been summarized. The first part present the distribution, type, morphology and way of impulse transmission as well as the role of the pulpal nerves. The second part present the growth of the nerve fibers in the dental structures. Through the analysis of all the research done on this subjects two important thighs can be deduced: the developing innervation correlates with the stage of the development of individual teeth rather than the chronological age of the animal, and the dental pulp compared to other structures is relatively late in maturing. This is why the axons don't penetrate into the dental papilla until crown formation commences. Then the probable role of the pulpal nerves in tooth growth (dentinogenesis) has been analysed, but this activity, even though unessential, is evident only in developing final phases and not in the early ones; this is why the axons are separated from the early growing tissues. Concluding, our review shows the variation in number, type and function of the pulpal nerves from their developing period to their complete formation until senescence.

Dental Pulp↗

Ontogenesis of olivocerebellar relationships. I. Studies by intracellular recordings of the multiple innervation of Purkinje cells by climbing fibers in the developing rat cerebellum.

The establishment of the adult innervation of Purkinje cells (PCs) by climbing fibers (CFs) was studied in the cerebellar vermis of the developing rat. Excitatory postsynaptic potentials (EPSPs) evoked in PCs by activation of the climbing fibers (CF-EPSPs) were recorded intracellularly from a total of 310 cells in young rats aged from 3 to 15 postnatal days. The CF system was activated by electrical stimulation of either the inferior olive (IO) nucleus or the region near the fastigial nucleus (juxtafastigial or JF stimulation). A given PC at each age was considered to be innervated by more than one CF when the amplitude of the spontaneous or evoked CF-EPSPs fluctuated in a stepwise manner. On the other hand, innervation of a PC by a single CF was established on the basis of the all-or-none character of CF-EPSPs. Two parameters were followed throughout development, the percentage of multiply innervated PCs and the mean number of steps in the evoked CF-EPSPs. The data presented confirm the transient multiple innervation of PCs by CFS on postnatal days 8 and 9 (Crépel, F., J. Mariani, and N. Delhaye-Bouchaud (1976) J. Neurobiol. 7: 567-578) and strongly suggest its existence at earlier stages (from postnatal day 3). Moreover, it is shown that the multiple innervation was maximal on postnatal day 5 and then decreased until the innervation by a single CF was established on day 15.

Action Potentials↗

Abnormal targeting of developing hippocampal mossy fibers after epileptiform activities via L-type Ca2+ channel activation in vitro.

The hippocampal mossy fibers, which originate from the dentate granule cells, develop mainly in the early postnatal period and are involved in numerous pathological processes. In this study, hippocampal slices prepared from premature rats were cultivated in the presence of convulsants to evaluate the influences of epileptiform activities on mossy fiber ontogeny. Electrophysiological and histochemical analyses revealed that prolonged hyperexcitability inhibited proper growth of the mossy fibers and caused ectopic innervation to the stratum oriens and the dentate molecular layer. These phenomena were prevented by pharmacological blockade of L-type Ca2+ channels, which did not affect convulsant-evoked ictal bursts. After single-pulse stimulation of the stratum granulosum in the slices cultured under paroxysmal conditions, the dentate gyrus displayed excessive excitation, but synaptic transmission to the CA3 region was hypoactive. However, brief repetitive stimulation elicited delayed epileptiform discharges in the CA3 region that were inhibited by an NMDA receptor antagonist. Chronic treatment with an L-type Ca2+ channel blocker ameliorated such aberrant neurotransmissions. These results suggest that ictal neuron activities at the developmental stage of the mossy fibers bring about the errant maturation associated with hippocampal dysfunction, which may form a cellular basis for the sequelae of childhood epilepsy, including chronic epilepsy or cognitive deficits. Thus I propose that L-type Ca2+ channel blockers can ameliorate the aversive prognosis of childhood epilepsy.

Animals↗

Differentiation of the brain stem structures in the salamander, Hynobius nebulosus.

Differentiation of the internal structure of the brain stem was analyzed in the salamander with special reference to neurons distributed in the marginal layer. It was found that the salamander brain stem was at first composed exclusively of the mantle layer. The marginal layer later differentiated peripherally. In these developmental stages, the mantle and marginal layers were clearly differentiated: the former was made up exclusively of the somata, while the latter was composed mainly of nerve fibers. As the development proceeded, these organization patterns were modified: a few cells migrated into the marginal layer. Cells migrating into the marginal layer formed various nuclei and layers such as the raphe nuclei, reticular formation and superficial cellular layers of the optic tectum. In later development stages, fibers in the marginal layer were myelinated, and neurons in the marginal layer were observed to become embedded among numerous myelinated fibers. Cytologically, the majority of neurons in early developmental stages were unipolar, extending a process peripherally into the marginal layer. In later developmental stages, neurons in a deep zone of the mantle layer remained unipolar, whereas those in the marginal layer and in the superficial zone of the mantle layer differentiated into multipolar cells. Thus, (1) the marginal layer differentiated peripherally as a cell free region; (2) cells in the mantle layer later migrated into the marginal layer, changing into multipolar neurons; (3) cells in the marginal layer formed reticular formation as well as various nuclei and layers in the peripheral white matter; and (4) as development proceeded, fibers in the marginal layer became myelinated.

Aging↗

Suppression of sucrose synthase gene expression represses cotton fiber cell initiation, elongation, and seed development.

Cotton is the most important textile crop as a result of its long cellulose-enriched mature fibers. These single-celled hairs initiate at anthesis from the ovule epidermis. To date, genes proven to be critical for fiber development have not been identified. Here, we examined the role of the sucrose synthase gene (Sus) in cotton fiber and seed by transforming cotton with Sus suppression constructs. We focused our analysis on 0 to 3 days after anthesis (DAA) for early fiber development and 25 DAA, when the fiber and seed are maximal in size. Suppression of Sus activity by 70% or more in the ovule epidermis led to a fiberless phenotype. The fiber initials in those ovules were fewer and shrunken or collapsed. The level of Sus suppression correlated strongly with the degree of inhibition of fiber initiation and elongation, probably as a result of the reduction of hexoses. By 25 DAA, a portion of the seeds in the fruit showed Sus suppression only in the seed coat fibers and transfer cells but not in the endosperm and embryo. These transgenic seeds were identical to wild-type seeds except for much reduced fiber growth. However, the remaining seeds in the fruit showed Sus suppression both in the seed coat and in the endosperm and embryo. These seeds were shrunken with loss of the transfer cells and were <5% of wild-type seed weight. These results demonstrate that Sus plays a rate-limiting role in the initiation and elongation of the single-celled fibers. These analyses also show that suppression of Sus only in the maternal seed tissue represses fiber development without affecting embryo development and seed size. Additional suppression in the endosperm and embryo inhibits their own development, which blocks the formation of adjacent seed coat transfer cells and arrests seed development entirely.

Cell Division↗

Transient expression of NMDA receptors during rearrangement of AMPA-receptor-expressing fibers in the developing inner ear.

A major reorganization of afferent and efferent nerve terminals, concomitant to significant neuronal cell loss and pruning of superfluous fibers, takes place during the development of the organ of Corti, prior to the onset of hearing. We examined the spatio/temporal distribution of subtype-specific AMPA- and N-methyl-d-aspartate (NMDA)-selective glutamate receptor proteins in postnatal inner ears from rats during this critical period. From the first postnatal day onwards, GluR2/3 receptor subtypes appeared in nerve endings of afferent fibers associated with inner and outer hair cells. During the following 2 weeks, GluR2/3 receptors were downregulated in exchange for GluR4 receptors. In parallel efferents projecting from the medial olivocochlear complex to the outer hair cells underwent synaptogenesis and efferents projecting from the lateral olivocochlear complex to the inner hair cells appeared to change contacts to the dendrites of afferents. Concomitant to these events, NMDA receptor subtypes NR1 and NR2A transiently appeared in hair cells as well as afferent and efferent fibers. Recently, we described a temporary expression of the neurotrophin receptor trkB in hair cells, coincident to the growth (GAP-43) and synaptogenesis (synaptophysin) of efferents. Here, we show that trkB was expressed together with NR1 receptors in hair cells in high spatio/temporal correlation with the rearrangement of afferents and efferents. Cochlea NMDA receptors may, therefore, be a part of the mechanism by which, in addition to neurotrophic activity, the mature phenotype of cochlea neurons is acquired through activity-dependent processes.

Animals↗

Rigor tension development in glycerinated rabbit psoas fibers at high salt concentrations.

We attempted to measure the rigor tension development by glycerinated fibers of rabbit psoas at high salt concentrations such as 0.5 M KCl. The measurements were made feasible by covalently crosslinking the rod-portion of thick filaments in the fibers in the rigor state with a water-soluble carbodiimide (EDC) so that the thick filaments are not dissolved even at 0.5 M KCl. EDC crosslinks, though with much a slower rate, the myosin cross-bridge heads to the thin filaments. At high salt concentrations, the fibers developed no active tension but developed rigor tension when they were put into a rigor solution from a contracting or relaxing solution. Removal of only Mg++ from a MgATP-containing solution induced similar rigor tension development. The magnitude of the rigor tension was proportional to the fraction of the cross-bridge heads that were crosslinked to the thin filaments. The results suggest that the rigor tension at high salt concentrations is generated by structural changes in the cross-bridge heads that are crosslinked to the thin filaments, when these heads release MgATP or Mg++ (with ATP retained) from their active sites, but not generated by re-formation of the rigor complexes of uncrosslinked myosin heads with the thin filaments. Extrapolation to 100% crosslinked heads gave an estimate of the rigor tension development of more than 1 kg wt/cm2 at high salt concentrations.

Actin Cytoskeleton↗

Innervation regulates myosin heavy chain isoform expression in developing skeletal muscle fibers.

The influence of innervation on primary and secondary myogenesis and its relation to fiber type diversity were investigated in two specific wing muscles of quail embryo, the posterior (PLD) and anterior latissimus dorsi (ALD). In the adult, these muscles are composed almost exclusively of pure populations of fast and slow fibers, respectively. When slow ALD and fast PLD muscles developed in ovo in an aneurogenic environment induced after neural tube ablation, the cardiac ventricular myosin heavy chain (MHC) isoform was not expressed. The adult slow MHC isoform, SM2, appeared by embryonic day 7 (ED 7) in normal innervated slow ALD but was not expressed in denervated muscle. Analysis of in vitro differentiation of myoblasts from fast PLD and slow ALD muscles isolated from ED 7 control and neuralectomized quail embryos showed no fundamental differences in the pattern of MHC isoform expression. Newly differentiated fibers accumulated cardiac ventricular, embryonic fast, slow SM1 and SM3 MHC isoforms. Nevertheless, the expression of slow SM2 isoform in myotubes formed from slow ALD myoblasts only occurred when myoblasts were cultured in the presence of embryonic spinal cord. Our studies demonstrate that the neural tube influences primary as well as secondary myotube differentiation in avian forelimb and facilitates the expression of different MHC, particularly slow SM2 MHC gene expression in slow myoblasts.

Acetylcholinesterase↗

Effects of hypothyroidism on the detyrosylation of alpha-tubulin in the parallel fibers of the developing rat cerebellar cortex.

The immunocytochemical staining of tyrosylated alpha-tubulin using the specific monoclonal antibody YL 1/2 decreases dramatically in the parallel fibers of the rat cerebellum during development. Detyrosylation starts at 14 days in the deeper part of the molecular layer and then spreads over the entire molecular layer within a week. Thyroid deficiency induces a marked delay in alpha-tubulin detyrosylation in the parallel fibers. Detyrosylation begins only at the age of 28 days in hypothyroid animals. Immunobinding assays of detyrosylated and tyrosylated alpha-tubulin concentrations in the cerebellar supernatant of developing rats confirm that thyroid deficiency increases the tyrosylated alpha-tubulin content at 21 and 35 days. Short-term thyroxine treatment (daily from day 8, 10 or 12, to day 14) do not induce the normal pattern of tubulin detyrosylation starting at 14 days, although they do induce parallel fiber lengthening. Daily treatment from birth is required to produce the alpha-tubulin modification found in normal rats. We propose that the microtubule detyrosylation which occurs during axonal maturation is not a prerequisite for axon growth and synaptic differentiation, but may rather be implicated in the stabilization of microtubules in the mature axons.

Aging↗

Destruction of different fiber tracts underlies development of lateral hypothalamic lesion-induced hyperthermia and loss of bombesin-induced hypothermia.

The relative roles of lateral hypothalamic cell bodies and fibers of passage were assessed in the development of lesion-induced hyperthermia and bombesin-induced hypothermia. Electrolytic lesions or discrete fiber transections were combined with intracisternal bombesin injection to show that each of these two thermoregulatory effects involves fibers crossing the borders of the lateral hypothalamus; however, the two effects primarily involve fibers crossing different borders. Thus, the hyperthermia and the abolition of bombesin-induced hypothermia which follow lateral hypothalamic damage appear to result from disruption of separate thermoregulatory pathways.

Animals↗

Amygdala kindling develops without mossy fiber sprouting and hippocampal neuronal degeneration in rats.

Repeated electrical stimulation of limbic structures has been reported to produce the kindling effect together with morphological changes in the hippocampus such as mossy fiber sprouting and/or neuronal loss. However, to argue against a causal role of these neuropathological changes in the development of kindling-associated seizures, we examined mossy fiber sprouting in amygdala (AM)-kindled rats using Timm histochemical staining, and evaluated the hippocampal neuronal degeneration in AM-kindled rats by terminal deoxynucleotidyl transferase-mediated digoxigenin-11-dUTP nick end labelling (TUNEL). Amygdala kindling was established by 10.3 +/- 0.7 electrical stimulations, and no increase in Timm granules (neuronal sprouting) was observed up to the time of acquisition of a fully kindled state. However, the density and distribution of Timm granules increased significantly in the dentate gyrus compared with unkindled rats after 29 after-discharges or more than 10 kindled convulsions. In addition, no significant increase in TUNEL-positive cells was found in the hilar polymorphic neurons or in CA3 pyramidal neurons of the kindled rats that had fewer than 29 after-discharges. However, a significant increase of TUNEL-positive cells was found in the granule cell layer in the dentate gyrus of the stimulated side after 18 after-discharges or 10 kindled convulsions. Our result show that AM kindling develops without evidence of mossy fiber sprouting, and that mossy fiber sprouting may appear after repeated kindled convulsions, following death of the granule cells in the dentate gyrus.

Amygdala↗