Sympathetic and Sensory Axons Invade the Brains of Nerve Growth Factor Transgenic Mice in the Absence of p75NTRExpression

doi:10.1006/exnr.1997.6664

Experimental Neurology

Volume 149, Issue 1, January 1998, Pages 284-294

https://doi.org/10.1006/exnr.1997.6664 Get rights and content

Abstract

Collateral sprouting, a nerve growth factor (NGF)-mediated growth response of undamaged peripheral axons, can be divided into reparative and aberrant axonal growth. We have previously shown that aberrant growth occurs in transgenic mice overexpressing NGF centrally under the control of the glial fibrillary acidic protein promoter. Both sympathetic and sensory fibers, stained immunohistochemically for tyrosine hydroxylase and calcitonin gene-related peptide, respectively, invade the cerebellum of postnatal transgenic mice, whereas no such axons are seen in age-matched wild-type cerebellum. Recent examination of mice possessing a null mutation for p75^NTRhas suggested that axon growth may be influenced by the functional expression of this receptor. To address the potential role of p75^NTRin axon growth, we have generated a new line of hybrid mice overexpressing NGF but lacking functional p75^NTRexpression. Postnatal (day 14) hybrid cerebellum possessed fewer aberrant sensory and sympathetic fibers compared to their age-matched transgenic counterparts. By adulthood, however, hybrid cerebellum displayed a robust plexus of axons stained immunohistochemically for calcitonin gene-related peptide and tyrosine hydroxylase. No neuronal or nonneuronal localization of p75^NTR-immunoreactive elements was observed in postnatal and adult hybrid cerebellum. Interestingly, sympathetic axons within the hybrid cerebellum displayed a markedly reduced axon density and staining intensity for NGF, suggesting a possible alteration in axonal sequestration of NGF. These results show that p75^NTRis not vital for new growth of NGF-sensitive sympathetic and sensory axons and that immunohistochemical detection of NGF at sympathetic axons requires the functional expression of p75^NTR.

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Proteomic assessment of sympathetic ganglia from adult mice that possess null mutations of ExonIII or ExonIV in the p75 neurotrophin receptor gene
2009, Brain Research
Citation Excerpt :
Second, densities of sympathetic axons in target organs of p75(III)−/− mice may be increased (e.g., in the submandibular salivary gland; Jahed and Kawaja, 2005), decreased (e.g., in the pineal gland; Lee et al., 1994), or unchanged (e.g., in the heart and urinary bladder; Jahed and Kawaja, 2005). Third, in the absence of p75NTR expression, the accumulation of retrogradely-derived NGF in trkA-positive somata (as identified by immunostaining) is markedly depleted, as compared to NGF immunostaining seen in sympathetic neurons of wild type mice (Coome et al., 1998; Walsh et al., 1999a,b; Krol et al., 2000). Parenthetically, there is no evidence as to whether the phenotype of ganglionic Schwann cells or satellite cells, which are found in immediate proximity to post-ganglionic sympathetic neurons, is affected in p75(III)−/− mice.
Neurotrophins, such as nerve growth factor (NGF), are capable of binding to the transmembrane p75 neurotrophin receptor (p75NTR), which regulates a variety of cellular responses including apoptosis and axonal elongation. While the development of mutant mouse strains that lack functional p75NTR expression has provided further insight into the importance of this neurotrophin receptor, there remains a paucity of information concerning how the loss of p75NTR expression may alter neural phenotypes. To address this issue, we assessed the proteome of the cervical sympathetic ganglia from two mutant lines of mice, which were compared to the ganglionic proteome of age-matched wild type mice. The ganglionic proteome of mice possessing two mutant alleles of either exonIII or exonIV for the p75NTR gene displayed detectable alterations in levels of Lamin A, tyrosine hydroxylase, and Annexin V, as compared to ganglionic proteome of wild type mice. Decreased expression of the basic isoform of tyrosine hydroxylase may be linked to perturbed NGF signaling in the absence of p75NTR in mutant mice. Stereological measurement showed significant increases in the number of sympathetic neurons in both lines of p75NTR-deficient mice, relative to wild type mice. This enhanced survival of sympathetic neurons coincides with shifts toward the more basic isoforms of Annexin V in mutant mice. This study, in addition to providing the first comparative proteomic assessment of sympathetic ganglia, sheds new light onto the phenotypic changes that occur as a consequence of a loss of p75NTR expression in adult mice.
Null mutations for exon III and exon IV of the p75 neurotrophin receptor gene enhance sympathetic sprouting in response to elevated levels of nerve growth factor in transgenic mice
2006, Experimental Neurology
Under normal conditions, expression of the p75 neurotrophin receptor (p75NTR) by sympathetic neurons can increase the affinity of the signaling receptor, trkA, to target-derived nerve growth factor (NGF) at distal axons. We have previously reported that sprouting of sympathetic axons into NGF-rich target tissues is enhanced when p75NTR expression is perturbed, leading to the postulate that p75NTR may restrain sympathetic sprouting in response to elevated NGF levels. These observations were made using mice having a null mutation of the third p75NTR exon, a line that may express a hypomorphic form of this receptor. Since mice carrying a null mutation of the fourth p75NTR exon may not express a similar splice variant, we sought to determine whether these animals possess the same phenotype of enhanced sympathetic sprouting in response to elevated levels of NGF. Both lines of transgenic mice lacking p75NTR displayed similar degrees of sympathetic axonal sprouting into the cerebellum and trigeminal ganglia, two target tissues having elevated levels of NGF protein. Furthermore, the densities of sympathetic axons in both targets were significantly greater than those observed in age-matched NGF transgenic siblings expressing full-length p75NTR. Our new findings provide a comparative analysis of the phenotype in two independent mutations of the same neurotrophin receptor, revealing that p75NTR plays an important role in restricting sympathetic sprouting in response to higher NGF levels.
The influences of p75 neurotrophin receptor and brain-derived neurotrophic factor in the sympathetic innervation of target tissues during murine postnatal development
2005, Autonomic Neuroscience: Basic and Clinical
Post-ganglionic sympathetic neurons express the p75 neurotrophin receptor (p75NTR) and brain-derived neurotrophic factor (BDNF), which together have been implicated in controlling the degree of efferent innervation of peripheral organs [Kohn, J., Aloyz, R.S., Toma, J.G., Haak-Frendscho, M., Miller, F.D. 1999. Functionally Antagonistic Interactions between the TrkA and p75 Neurotrophin Receptors Regulate Sympathetic Neuron Growth and Target Innervation. J. Neurosci. 19, 5393–5408]. To examine this concept further, we developed null mutant mice lacking both p75NTR and BDNF, and assessed whether the loss of this receptor–ligand interaction negatively impacts the degree of sympathetic innervation to various target tissues. Between postnatal days 10 and 14, hearts, urinary bladders, kidneys, and submandibular salivary glands were isolated from p75^−/−/BDNF^−/−, p75^−/−, BDNF^−/−, and wild type siblings. Sympathetic axons were visualized using tyrosine hydroxylase (TH) immunohistochemistry, and TH protein levels were quantified by immunoblotting. Concerning the sympathetic innervation of the heart, urinary bladder and kidneys, no differences were seen in single and double null mutant mice, as compared with their wild type siblings. Sympathetic innervation of the submandibular salivary gland was, however, increased in both p75^−/− and p75^−/−/BDNF^−/− mice over control mice. These results reveal that an absence of p75NTR and/or BDNF expression does not perturb the degree of sympathetic innervation of many peripheral tissues during postnatal development, and that a lack of p75NTR expression may actually enhance the density of these efferent fibers in other target tissues, such as the salivary glands.
Nerve growth factor alters p75 neurotrophin receptor-induced effects in mouse facial motoneurons following axotomy
2002, Brain Research
Citation Excerpt :
We have previously documented that post-ganglionic sympathetic axons sprout into the brains of NGF/p75+/+ and NGF/p75−/− mice [18,32]. In response to facial nerve axotomy, increased expression of GFAP among reactive astrocytes causes a concomitant increase in ectopic NGF protein synthesis in both NGF/p75+/+ and NGF/p75−/− mice [8]. Twenty-five days following a unilateral facial nerve transection, numerous sympathetic axons stained immunohistochemically for TH were seen in the axotomized nucleus of NGF/p75+/+ and NGF/p75−/− mice but not in wild type mice; no immunostained axons were evident in the contralateral nucleus of these animals (Fig. 2).
The p75 neurotrophin receptor (p75^NTR) has been implicated as being detrimental for cell survival in facial motoneurons following injury. Although facial motoneurons do not respond to nerve growth factor (NGF) under normal circumstances, this study shows that NGF can interfere with p75^NTR-mediated cell survival effects on motoneurons following injury. Twenty-five days following injury, the proportion of surviving axotomized neurons in NGF/p75^+/+ mice, which overexpress NGF, was significantly higher compared to wild-type mice, while NGF/p75^−/− mice, which overexpress NGF but carry two mutated alleles for p75^NTR, had fewer neurons compared to wild-type and p75^−/− mice, which carry two mutated alleles for p75^NTR, resulting in a lack of functional expression of this receptor. Sympathetic axons sprouted into the axotomized facial nucleus of both NGF/p75^+/+ and NGF/p75^−/− following injury, due to transgene expression of NGF in reactive astrocytes. Removal of these sympathetic axons enhanced the number of surviving axotomized neurons in NGF/p75^−/− mice but not in NGF/p75^+/+ mice. Although motoneurons do not express trkA and should therefore be unresponsive to NGF, our results reveal that NGF can influence p75-mediated motoneuron survival following axotomy.
p75 and TrkA receptors are both required for uptake of NGF in adult sympathetic neurons: Use of a novel fluorescent NGF conjugate
2001, Brain Research
Citation Excerpt :
Moreover, p75NTR has been shown to cooperate with TrkA in the differentiation of MAH cells, a neuronal precursor cell line [70] and in modulating the internalization of NGF in mutant Sf9 cells [30]. Finally, it has recently been demonstrated that p75NTR expression is required for the detection of NGF in sympathetic axons [16,36] and that sympathetic and sensory innervation of the meningeal arteries is severely perturbed in p75NTR-deficient mice [48]. A role for p75NTR has also been demonstrated in mediating uptake of NT-4 and BDNF in peripheral neurons of adult rats [19].
We have developed and tested the biological activity and specificity of a novel fluorescent dextran-Texas Red–nerve growth factor (DTR–NGF) conjugate. DTR–NGF was found to promote survival and neurite outgrowth in cultured dissociated sympathetic neurons similarly to native NGF. The conjugate was taken up and transported retrogradely by terminal sympathetic nerves innervating the iris to neurons in the ipsilateral superior cervical ganglion (SCG) of young adult rats. Uptake and transport was assessed by counting numbers of labelled neurons and by measuring intensity of neuronal labelling using confocal microscopy and image analysis. DTR–NGF labelling in SCG neurons was shown to be dose-dependent with an EC₅₀ of 75 ng. Similar concentrations of unconjugated DTR resulted in no neuronal labelling. DTR–NGF uptake was competed off using a 50-fold excess of native NGF, resulting in a 73% reduction in numbers of labelled neurons. Pretreatment of nerve terminals with function-blocking antibodies against the low (p75) and high (TrkA) affinity NGF receptors resulted in a large (85–93%) reduction in numbers of DTR–NGF labelled neurons. Anti-p75 and anti-TrkA antibodies had comparable effects which were concentration-dependent. These findings indicate that both receptors are required for uptake of NGF in adult rat sympathetic neurons. In particular, the results provide strong evidence that the p75 receptor plays a more active role in transducing the NGF signal than has been proposed.
Localization of low affinity nerve growth factor receptor in the rat inferior olivary complex during development and plasticity of climbing fibres
2001, Developmental Brain Research
The rat olivocerebellar pathway has a precise topography from an inferior olive (IOC) to Purkinje cells in the contralateral hemicerebellum. While its development and plasticity have been documented, the molecular mechanisms underlying these events are not fully elucidated. Neurotrophins are a family of growth factors with diverse roles in development and neuronal plasticity, acting through a two-receptor system, including a low affinity receptor (LNGFR) which binds all neurotrophins with similar affinity. Since neurotrophins are present in the cerebellum during early postnatal development when LNGFR is synthesized in the IOC, they may act as target-derived trophic agents for climbing fibres during development and plasticity. To assess this, standard immunohistochemistry was used to document the distribution of LNGFR in the rat IOC during climbing fibre development and until cerebellar development was complete at postnatal day 28 (P28). LNGFR immunoreactivity (LNGFR-IR) was detected in the IOC from P0 until P15, however after P7 it diminished in intensity and distribution, a change which indicates a relationship between cerebellar neurotrophins and climbing fibre development. After denervation of the left hemicerebellum, there was an apparent increase in inferior olivary LNGFR-IR that was concurrent with climbing fibre re-innervation. Thus the results of this study support the hypothesis that neurotrophins are involved in climbing fibre development and suggest a possible contribution to the plasticity of the olivocerebellar pathway.

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^☆: C. W. Cotman

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Regular ArticleSympathetic and Sensory Axons Invade the Brains of Nerve Growth Factor Transgenic Mice in the Absence of p75NTRExpression☆

Abstract

Brain Res.

Neuron

Neuron

TINS

Neurosci. Lett.

Brain Res.

Exp. Neurol.

Cell

Prog. Brain Res.

J. Biol. Chem.

Dev. Biol.

Neuron

Dev. Biol.

Brain Res.

Nerve growth factor and its low-affinity receptor promote Schwann cell migration

Proc. Natl. Acad. Sci. USA

The low-affinity p75 growth factor (NGF) receptor mediates NGF-induced tyrosine phosphorylation

Proc. Natl. Acad. Sci. USA

Distribution of nerve growth factor-like immunoreactive neurons in the adult rat brain following colchicine treatment

J. Comp. Neurol.

Nonregenerative axonal growth within the mature mammalian brain: Ultrastructural identification of sympathohippocampal sprouts

J. Neurosci.

Detection of NGF-like activity in human brain tissue: Increased levels in Alzheimer's disease

J. Neurosci.

Evidence that endogenous β nerve growth factor is responsible for the collateral sprouting, but not the regeneration, of nociceptive axons in adult rats

Proc. Natl. Acad. Sci. USA

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

J. Comp. Neurol.

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

J. Comp. Neurol.

High-affinity NGF binding requires coexpression of the trk proto-oncogene and the low-affinity NGF receptor

Nature

Regular Article
Sympathetic and Sensory Axons Invade the Brains of Nerve Growth Factor Transgenic Mice in the Absence of p75^NTRExpression☆