GAP-43 (B-50) and C-Jun Are Up-Regulated in Axotomized Neurons of Clarke's Nucleus after Spinal Cord Injury in the Adult Rat

doi:10.1006/nbdi.1998.0231

Neurobiology of Disease

Volume 6, Issue 2, April 1999, Pages 122-130

https://doi.org/10.1006/nbdi.1998.0231 Get rights and content

Abstract

The growth-associated protein GAP-43 (B-50) and the transcription factor C-Jun are involved in regeneration of the injured nervous system. In this study, we investigated the possibility of the induction of GAP-43 and C-Jun in axotomized neurons of Clarke's nucleus (CN) in adult rats, of which a large population undergoes degeneration several weeks after a low thoracic lateral funiculotomy of the spinal cord.In situhybridization and immunohistochemistry revealed a transient up-regulation of GAP-43 mRNA, C-Jun protein, and its activated, phosphorylated form, peaking around 7 days after injury in preferentially large diameter CN-neurons ipsilateral and caudal to the lesion. Our results document that some populations of axotomized central nervous system neurons, similar to axotomized regenerating neurons of the peripheral nervous system, can up-regulate GAP-43 and C-Jun, even if they are destined to degenerate. This might reflect a transient regenerative capacity, which fails over time.

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Cited by (32)

Up-regulation of GAP-43 in the chinchilla ventral cochlear nucleus after carboplatin-induced hearing loss: Correlations with inner hair cell loss and outer hair cell loss
2013, Hearing Research
Citation Excerpt :
The up-regulation of GAP-43 in auditory nerve fibers on the treated side is indicative of mechanisms associated with regeneration or remodeling. On the other hand, surviving neurons showing markers for regeneration or remodeling such as GAP-43 up-regulation may nevertheless degenerate at a later time point (Schmitt et al., 1999; Kraus and Illing, 2005). In addition to rapid degeneration, Takeno et al. (1998) showed slow progressive degeneration of spiral ganglion neurons between 2 and 12 weeks after carboplatin treatment, where spiral ganglion cell death and IHC loss matched in extent as well as in location on the cochlear axis.
Inner ear damage leads to nerve fiber growth and synaptogenesis in the ventral cochlear nucleus (VCN). In this study, we documented the relationship between hair cell loss patterns and synaptic plasticity in the chinchilla VCN using immunolabeling of the growth associated protein-43 (GAP-43), a protein associated with axon outgrowth and modification of presynaptic endings. Unilateral round window application of carboplatin caused hair cell degeneration in which inner hair cells (IHC) were more vulnerable than outer hair cells (OHC). One month after carboplatin treatment (0.5–5 mg/ml), we observed varying patterns of cochlear hair cell loss and GAP-43 expression in VCN. Both IHC loss and OHC loss were strongly correlated with increased GAP-43 immunolabeling in the ipsilateral VCN. We speculate that two factors might promote the expression of GAP-43 in the VCN; one is the loss of afferent input through IHC or the associated type I auditory nerve fibers. The other occurs when the medial olivocochlear efferent neurons lose their cochlear targets, the OHC, and may as compensation increase their synapse numbers in the VCN.
Relationship between noise-induced hearing-loss, persistent tinnitus and growth-associated protein-43 expression in the rat cochlear nucleus: Does synaptic plasticity in ventral cochlear nucleus suppress tinnitus?
2011, Neuroscience
Citation Excerpt :
In eight of nine rats, GAP-43 was up-regulated in the auditory nerve on the deafened side. GAP-43 in fibers is typically associated with regeneration, but it is also expressed in injured neurons that attempt to regenerate but eventually die (Schmitt et al., 1999; Kraus and Illing, 2005). Noise-induced auditory nerve degeneration has been previously reported (Ylikoski et al., 1998; Michler and Illing, 2003).
Aberrant, lesion-induced neuroplastic changes in the auditory pathway are believed to give rise to the phantom sound of tinnitus. Noise-induced cochlear damage can induce extensive fiber growth and synaptogenesis in the cochlear nucleus, but it is currently unclear if these changes are linked to tinnitus. To address this issue, we unilaterally exposed nine rats to narrow-band noise centered at 12 kHz at 126 dB sound pressure level (SPL) for 2 h and sacrificed them 10 weeks later for evaluation of synaptic plasticity (growth-associated protein 43 [GAP-43] expression) in the cochlear nucleus. Noise-exposed rats along with three age-matched controls were screened for tinnitus-like behavior with gap prepulse inhibition of the acoustic startle (GPIAS) before, 1–10 days after, and 8–10 weeks after the noise exposure. All nine noise-exposed rats showed similar patterns of severe hair cell loss at high- and mid-frequency regions in the exposed ear. Eight of the nine showed strong up-regulation of GAP-43 in auditory nerve fibers and pronounced shrinkage of the ventral cochlear nucleus (VCN) on the noise-exposed side, and strong up-regulation of GAP-43 in the medial ventral VCN, but not in the lateral VCN or the dorsal cochlear nucleus. GAP-43 up-regulation in VCN was significantly greater in Noise-No-Tinnitus rats than in Noise-Tinnitus rats. One Noise-No-Tinnitus rat showed no up-regulation of GAP-43 in auditory nerve fibers and only little VCN shrinkage, suggesting that auditory nerve degeneration plays a role in tinnitus generation. Our results suggest that noise-induced tinnitus is suppressed by strong up-regulation of GAP-43 in the medial VCN. GAP-43 up-regulation most likely originates from medial olivocochlear neurons. Their increased excitatory input on inhibitory neurons in VCN may possibly reduce central hyperactivity and tinnitus.
Inhibiting epidermal growth factor receptor attenuates reactive astrogliosis and improves functional outcome after spinal cord injury in rats
2011, Neurochemistry International
Citation Excerpt :
As shown in Fig. 4D, the area of demyelination region in the AG1478-treated group was significant smaller comparing with that of control group. It has been suggested that the absence of significant regeneration by intrinsic neurons of the central nervous system (CNS) is correlated with a failure to up-regulate GAP-43 (Schmitt et al., 1999). Hence, we tried to investigate whether EGFR inhibition had any effect on GAP-43 expression after SCI.
As a physical barrier to regenerating axons, reactive astrogliosis is also a biochemical barrier which can secrete inhibitory molecules, including chondroitin sulfate proteoglycans (CSPGs) in the pathological mechanism of spinal cord injury (SCI). Thus, inhibition of astroglial proliferation and CSPG production might facilitate axonal regeneration after SCI. Recent studies have demonstrated that epidermal growth factor receptor (EGFR) activation triggers quiescent astrocytes into becoming reactive astrocytes and forming glial scar after CNS injury. In the present study, we investigated whether a specific EGFR inhibitor (AG1478) could attenuate the reactive astrogliosis and production of CSPGs, alleviate demyelination, and eventually enhance the functional recovery after SCI in rats. Our results showed that pEGFR immunoreactivity was up-regulated significantly post injury, mainly confined to astrocytes. Meanwhile, astrocytes near the injury site after SCI became activated obviously characterized by hypertrophic morphology and enhanced GFAP expression. However, administration of AG1478 remarkably reduced trauma induced-reactive astrogliosis and accumulation of CSPGs. Furthermore, the treatment with AG1478 also alleviated demyelination, increased expression of growth-associated proteins-43 (GAP-43) and improved hindlimb function after SCI. Therefore, the local blockade of EGFR in an injured area is beneficial to functional outcome by facilitating a more favorable environment for axonal regeneration in SCI rats.
Investigating regeneration and functional integration of CNS neurons: Lessons from zebrafish genetics and other fish species
2011, Biochimica et Biophysica Acta - Molecular Basis of Disease
Citation Excerpt :
This effective gene knock-down technology is an economical and informative loss-of-function complement to tests in mammals, where L1 has been shown to be up-regulated during regeneration in CNS nerve grafts and L1 fusion protein application can promote functional recovery of locomotion [128,129]. Similarly, GAP-43 is up-regulated in zebrafish spinal cord regeneration [125], as in mammals [130–132]. On the other hand, another cell-adhesion molecule, protein zero (P0, a component of the myelin sheath) shows substantive difference between anamniotes with regenerative capacity when compared to mammals with limited spinal cord regenerative potential [85,133]: This and difference to mammals allow studies in zebrafish to contribute to understanding re-myelination following regeneration [133].
Zebrafish possess a robust, innate CNS regenerative ability. Combined with their genetic tractability and vertebrate CNS architecture, this ability makes zebrafish an attractive model to gain requisite knowledge for clinical CNS regeneration. In treatment of neurological disorders, one can envisage replacing lost neurons through stem cell therapy or through activation of latent stem cells in the CNS. Here we review the evidence that radial glia are a major source of CNS stem cells in zebrafish and thus activation of radial glia is an attractive therapeutic target. We discuss the regenerative potential and the molecular mechanisms thereof, in the zebrafish spinal cord, retina, optic nerve and higher brain centres. We evaluate various cell ablation paradigms developed to induce regeneration, with particular emphasis on the need for (high throughput) indicators that neuronal regeneration has restored sensory or motor function. We also examine the potential confound that regeneration imposes as the community develops zebrafish models of neurodegeneration. We conclude that zebrafish combine several characters that make them a potent resource for testing hypotheses and discovering therapeutic targets in functional CNS regeneration. This article is part of a Special Issue entitled Zebrafish Models of Neurological Diseases.
GM-CSF inhibits glial scar formation and shows long-term protective effect after spinal cord injury
2009, Journal of the Neurological Sciences
This study investigated the effects of granulocyte macrophage-colony stimulating factor (GM-CSF) on the scar formation and repair of spinal cord tissues in rat spinal cord injury (SCI) model.
Sprague–Dawley male rats (8 weeks old) were randomly divided into the sham-operated group, spinal cord injury group, and injury with GM-CSF treated group. A spinal cord injury was induced at T9/10 levels of rat spinal cord using a vascular clip. GM-CSF was administrated via intraperitoneal (IP) injection or on the dural surface using Gelfoam at the time of SCI. The morphological changes, tissue integrity, and scar formation were evaluated until 4 weeks after SCI using histological and immunohistochemical analyses.
The administration of GM-CSF either via IP injection or local treatment significantly reduced the cavity size and glial scar formation at 3–4 weeks after SCI. GM-CSF also reduced the expression of core proteins of chondroitin sulfate proteoglycans (CSPGs) such as neurocan and NG2 but not phosphacan. In particular, an intensive expression of glial fibriallary acidic protein (GFAP) and neurocan found around the cavity at 4 weeks was obviously suppressed by GM-CSF. Immunostaining for neurofilament (NF) and Luxol fast blue (LFB) showed that GM-CSF preserved well the axonal arrangement and myelin structure after SCI. The expression of GAP-43, a marker of regenerating axons, also apparently increased in the rostral grey matter by GM-CSF.
These results suggest that GM-CSF could enhance long-term recovery from SCI by suppressing the glial scar formation and enhancing the integrity of axonal structure.
Reconnecting neuronal networks in the auditory brainstem following unilateral deafening
2005, Hearing Research
When we disturbed the auditory input of the adult rat by cochleotomy or noise trauma on one side, several substantial anatomical, cellular, and molecular changes took place in the auditory brainstem. We found that: (1) cochleotomy or severe noise trauma both lead to a considerable increase of immunoreactivity of the growth-associated protein GAP-43 in the ventral cochlear nucleus (VCN) of the affected side; (2) the expression of GAP-43 in VCN is restricted to presynaptic endings and short fiber segments; (3) axon collaterals of the cholinergic medial olivocochlear (MOC) neurons are the path along which GAP-43 reaches VCN; (4) partial cochlear lesions induce the emergence of GAP-43 positive presynaptic endings only in regions tonotopically corresponding to the extent of the lesion; (5) judging from the presence of immature fibers and growth cones in VCN on the deafened side, at least part of the GAP-43 positive presynaptic endings appear to be newly formed neuronal contacts following axonal sprouting while others may be modified pre-existing contacts; and (6) GAP-43 positive synapses are formed only on specific postsynaptic profiles, i.e., glutamatergic, glycinergic and calretinin containing cell bodies, but not GABAergic cell bodies. We conclude that unilateral deafening, be it partial or total, induces complex patterns of reconnecting neurons in the adult auditory brainstem, and we evaluate the possibility that the deafness-induced chain of events is optimized to remedy the loss of a bilaterally balanced activity in the auditory brainstem.

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^☆: F. J. Seil

¹: To whom correspondence and reprint requests should be addressed. Fax: +49 241 8888444.E-mail: [email protected].

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Regular ArticleGAP-43 (B-50) and C-Jun Are Up-Regulated in Axotomized Neurons of Clarke's Nucleus after Spinal Cord Injury in the Adult Rat☆

Abstract

Exp. Neurol.

Neuroscience

Neuroscience

Mol. Brain Res.

Trends in Neurosci.

Neurosci. Lett.

Brain Res.

Neuroscience

Exp. Neurol.

Exp. Neurol.

Brain Res.

Neuroscience

Neuronal response to axotomy: Consequences and possibilities for rescue from permanent atrophy or cell death

Neural Regeneration and Transplantation

Distribution of B-50 (GAP-43) mRNA and protein in the normal adult human spinal cord

Acta Neuropathol.

C-Jun expression in adult root dorsal ganglion neurons: Differential response after central or peripheral axotomy

Exp. Neurol.

Intrinsic versus extrinsic factors in determining the regeneration of the central processes of rat dorsal root ganglion neurons: The influence of a peripheral nerve graft

J. Comp. Neurol.

Axon reaction in the red nucleus of the rat. Perikaryal volume changes and the time course of chromatolysis following cervical and thoracic lesions

Acta Neuropathol.

Altered gene expression in neurons during programmed cell death: Identification of C-Jun as necessary for neuronal apoptosis

J. Cell Biol.

Regular Article
GAP-43 (B-50) and C-Jun Are Up-Regulated in Axotomized Neurons of Clarke's Nucleus after Spinal Cord Injury in the Adult Rat☆