Biomedical subjects

A Gloster

Publications and source records attributed to A Gloster.

8 recordsLinked to original sources

PubMed1999

Early induction of Talpha1 alpha-tubulin transcription in neurons of the developing nervous system.

In this report, we have examined the relationship between the onset of neuronal gene transcription and neuronal development by characterizing expression of the early panneuronal Talpha1 alpha-tubulin promoter in developing neurons. In the peripheral nervous system, detectable expression of a beta-galactosidase transgene driven by the Talpha1 promoter (Talpha1:nlacZ) was coincident with neuronal birth dates, with the exception of sympathetic neuroblasts, which expressed the transgene prior to terminal mitosis. Similarly, in the central nervous system, the onset of beta-galactosidase expression was coincident with neuronal birth dates in most identifiable populations of central neurons. A small subpopulation of transgene-positive cells localized to ventricular zones, but the vast majority was observed in locations consistent with their identification as migrating and/or differentiating neurons. To determine more precisely the temporal relationship between transgene expression and terminal mitosis, we analyzed cultures of cortical progenitors that become postmitotic neurons in vitro. When initially plated, the vast majority of cells consisted of dividing, nestin-positive progenitors. Neurons differentiated from these progenitors as early as 1 day in vitro, as indicated by immunostaining for betaIII-tubulin, a neuron-specific tubulin isotype that is turned on shortly after terminal mitosis. Double-labeling studies showed that Talpha1:nlacZ expression was detectable in the same cells and at approximately the same time as was betaIII-tubulin, indicating that detectable transcription of the Talpha1 alpha-tubulin promoter commences at the time of terminal mitosis, at least in culture. This promoter, therefore, provides a valuable tool for genetic manipulation of early developing neurons in transgenic mice.

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PubMed1997

Transgenic mice expressing the intracellular domain of the p75 neurotrophin receptor undergo neuronal apoptosis.

We have asked whether p75(NTR) may play a role in neuronal apoptosis by producing transgenic mice that express the p75(NTR) intracellular domain within peripheral and central neurons. These animals showed profound reductions in numbers of sympathetic and peripheral sensory neurons as well as cell loss in the neocortex, where there is normally little or no p75(NTR) expression. Developmental loss of facial motor neurons was not observed, but induced expression of the p75(NTR) intracellular domain within adult animals led to increased motor neuron death after axotomy. Biochemical analyses suggest that these effects were not attributable to a p75(NTR)-dependent reduction in trk activation but instead indicate that the p75(NTR) intracellular domain may act as a constitutive activator of signaling cascades that regulate apoptosis in both peripheral and central neurons.

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PubMed1997

Transcriptional repression of the growth-associated T alpha1 alpha-tubulin gene by target contact.

In this report, we address the molecular mechanisms that regulate axonal growth by focusing on the gene for one of the major axonal cytoskeletal proteins, T alpha1 alpha-tubulin. During the developmental growth of sympathetic neurons, transcription of a beta-galactosidase transgene driven by the T alpha1 promoter (T alpha1:nlacZ) was high until the time of target innervation and neuronal maturation, when it decreased significantly. In mature animals, T alpha1:nlacZ transcription remained relatively low until target contact was experimentally disrupted; when facial motoneurons were axotomized, T alpha1:nlacZ transgene expression increased, was maximal for 1-7 days, and, if neurons regenerated and reinnervated their target musculature, returned to control levels by 49 days. In contrast, if regeneration and reestablishment of target contact were inhibited, transgene expression remained elevated. To determine whether this increased transcription was due to the loss of target contact or to axonal loss, we transected sympathetic neurons that project to the eye either close to or far from their cell bodies. In both cases, when target contact was severed, T alpha1:nlacZ transcription increased. These experiments indicate that transcription of the T alpha1 alpha-tubulin promoter is repressed by target contact in both developing and mature neurons. We suggest that this repression is due to a target-derived "stop-growth" factor that retrogradely signals to regulate transcription of this and other genes that are required for axonal growth.

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PubMed1997

Synaptic innervation density is regulated by neuron-derived BDNF.

In this report, we have examined the role of neuron-derived BDNF at an accessible synapse, that of preganglionic neurons onto their sympathetic neuron targets. Developing and mature sympathetic neurons synthesize BDNF, and preganglionic neurons express the full-length BDNF/TrkB receptor. When sympathetic neuron-derived BDNF is increased 2- to 4-fold in transgenic mice, preganglionic cell bodies and axons hypertrophy, and the synaptic innervation to sympathetic neurons is increased. Conversely, when BDNF synthesis is eliminated in BDNF -/- mice, preganglionic synaptic innervation to sympathetic neurons is decreased. Together these results indicate that variations in neuronal neurotrophin synthesis directly regulate neuronal circuitry by selectively modulating synaptic innervation density.

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PubMed1995

NGF-dependent and NGF-independent recovery of sympathetic function after chemical sympathectomy with 6-hydroxydopamine.

To help clarify the distinction between the nerve growth factor (NGF)-dependent collateral sprouting of sympathetic nerves and their NGF-independent regeneration after crush, we used 6-hydroxydopamine (OHDA) to destroy the sympathetic terminals in adult rats; this leaves the axons damage-free. Would recovery occur by regeneration and/or collateral sprouting? A single 6-OHDA injection abolished the sympathetic pilomotor field revealed by electrical stimulation of a cutaneous nerve. Recovery began within 2 days, and by 20 days the field was reestablished. If the field was "isolated" by adjacent denervations at the time of 6-OHDA treatment, the recovering pilomotor field expanded extensively into the surrounding territory. In the presence of anti-NGF, however, the pilomotor field expansion ceased at about 60% of its former size; if anti-NGF treatment was discontinued, expansion recommended and extended into the surrounding skin. We suggest that the latter, NGF-dependent, growth phase corresponds to collateral sprouting, and the initial NGF-independent one to regeneration. After simple nerve crush, however, such regeneration can triple the normal sympathetic field size. This difference between crush- and 6-OHDA-induced regeneration might relate to the "cell body reaction" (CBR); the CBR is reduced with increasing distance of the lesion and is undetectable after a 6-OHDA lesion. Since the CBR and the vigor of regeneration are both increased by repeated axonal injury, we tested the effects of multiple 6-OHDA treatments; this significantly increased the initial NGF-independent expansion. We hypothesize that regeneration is regulated largely by mechanisms associated with the CBR, and that neurotrophin-dependent collateral sprouting occurs independently of these.

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PubMed1994

Expression of NGF receptor and GAP-43 mRNA in DRG neurons during collateral sprouting and regeneration of dorsal cutaneous nerves.

The collateral sprouting of intact sensory axons and the regeneration of damaged ones differ in a number of respects. Regeneration is triggered by axotomy-induced damage, probably involves the loss of a peripheral signal, and appears to occur independently of NGF, while collateral sprouting is evoked and sustained by an increase in a target-derived signal, namely NGF. New findings strengthen the distinction between these two phenomena. Nerve growth factor receptor (NGFR) mRNA is increased in undamaged DRG neurons whose axons are sprouting into denervated skin. This response is related to an increased availability of target-derived NGF, a proposal supported by a number of findings including increased NGF mRNA in the denervated target. In contrast, we observed little or no change in the NGFR mRNA levels in regenerating neurons, consistent with the observations that NGF does not play a role in this process. However, increases in neuronal GAP-43 mRNA are found during both regeneration and collateral sprouting, a result in keeping with the proposal that GAP-43 is primarily associated with nerve growth, and the observation that GAP-43 expression is not especially influenced by NGF.

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PubMed1994

The T alpha 1 alpha-tubulin promoter specifies gene expression as a function of neuronal growth and regeneration in transgenic mice.

We have previously demonstrated that one member of the alpha-tubulin multigene family, termed T alpha 1 in rats, is regulated as a function of neuronal growth and regeneration. To elucidate the molecular mechanisms responsible for coupling gene expression to morphological differentiation, we have isolated the T alpha 1 gene, have fused 1.1 kb of the 5' flanking region to a nuclear lacZ reporter gene, and have generated transgenic mice. Analysis of these transgenic mice demonstrated that marker gene expression was specific to the CNS and PNS, with expression in vivo at embryonic day 13.5 being similar to expression of the endogenous gene. Moreover, the induction of transgene expression was correlated temporally with neuronal commitment in developing neural crest-derived peripheral neurons and in the developing retina. Immunocytochemical analysis of mixed primary embryonic brain cultures confirmed that transgene expression was specific to neurons, with the majority of neurons, but not astrocytes or oligodendrocytes, expressing beta-galactosidase. Transgene expression in vivo was maintained in developing neurons until early in postnatal life, subsequent to which its expression decreased coincident with neuronal maturation. The transgene was then reinduced in regenerating facial motoneurons following unilateral axotomy of the facial nerve. Thus, 1.1 kb of 5' flanking sequence from the T alpha 1 gene contains the sequence elements responsible for specifying gene expression to embryonic neurons and for subsequently regulating gene expression in both developing and mature neurons as a function of morphological growth.

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PubMed1992

Sympathetic nerves in adult rats regenerate normally and restore pilomotor function during an anti-NGF treatment that prevents their collateral sprouting.

We have used anti-nerve growth factor (anti-NGF) [corrected] administration to study the NGF dependency of the reinnervation of denervated skin by sympathetic nerves in the adult rat. Sympathetic pilomotor fields were revealed by electrical stimulation of selected dorsal cutaneous nerves; the affected skin rapidly assumed a "gooseflesh" appearance, sharply demarcated from surrounding unstimulated skin. Examined 2-5 days after section of neighboring nerves, the "isolated" pilomotor field of the spared nerve was found to be coextensive with an area of amine-fluorescent fibers that were associated with pilomotor muscles and blood vessels. After its isolation, a pilomotor field begins to expand into the surrounding deprived territory, reaching a maximum size at approximately 40 days. Fluorescence studies confirmed that new sympathetic fiber growth had occurred into the expanded regions of such fields. Daily injections of polyclonal anti-NGF serum completely prevented these pilomotor field expansions. Following termination of the anti-NGF treatment, expansion proceeded normally. Finally, if the onset of anti-NGF treatment was delayed until pilomotor field expansion had already commenced, further expansion was halted. Regeneration of sympathetic fibers was evoked by crushing a selected nerve. Recovery of pilomotor function in the totally denervated skin was first detected at about 20 days postcrush, and the field progressively enlarged over the next 40 days. Although the imposed NGF deprivation is known to cause a demonstrable shrinkage, and presumably atrophy, of sympathetic ganglia, the anti-NGF treatment appeared to impair neither the restoration of a pilomotor field after nerve crush, nor its continued expansion into skin regions well beyond that originally supplied by the nerve, i.e., into territory whose invasion by collateral sprouts would have been totally prevented by the treatment. During such NGF deprivation, fluorescent regenerating fibers were visualized in the nerve trunk. We conclude that even though the regenerating and collaterally sprouting sympathetic fibers probably utilise the same degenerating dermal pathways to reach and functionally reinnervate the same denervated targets, only the collateral sprouting of the uninjured axons is dependent upon endogenous NGF. These findings extend the results described earlier for nociceptive fibers, and suggest that the contrasting dependencies upon growth factors of sprouting and regeneration might apply throughout the adult nervous system.

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