RESUMEN

BACKGROUND: Conventional processing of nerve for histomorphometry is resource-intensive, precluding use in intraoperative assessment of nerve quality during nerve transfer procedures. Stimulated Raman scattering (SRS) microscopy is a label-free technique that enables rapid and high-resolution histology. METHODS: Segments of healthy murine sciatic nerve, healthy human obturator nerve, and human cross-facial nerve autografts were imaged on a custom SRS microscope. Myelinated axon quantification was performed through segmentation using a random forest machine learning algorithm in commercial software. RESULTS: High contrast, high-resolution imaging of nerve morphology was obtained with SRS imaging. Automated myelinated axon quantification from cross-sections of healthy human nerve imaged using SRS was achieved. CONCLUSIONS: Herein, we demonstrate the use of a label-free technique for rapid imaging of murine and human peripheral nerve cryosections. We illustrate the potential of this technique to inform intraoperative decision-making through rapid automated quantification of myelinated axons using a machine learning algorithm.

Asunto(s)

Nervio Facial/química , Nervio Obturador/química , Nervio Ciático/química , Espectrometría Raman/métodos , Animales , Nervio Facial/anatomía & histología , Humanos , Ratones , Microscopía Confocal/métodos , Nervio Obturador/anatomía & histología , Nervio Ciático/anatomía & histología

RESUMEN

A large number of genes regulate the process of regeneration after the nerve injury. Facial nerve axotomy of rats provides us with a useful model in which to study the molecular and cellular mechanisms of peripheral nerve regeneration. We performed the subtracted library screening of the facial nucleus after the facial nerve axotomy. We identified the sonic hedgehog (Shh) as a candidate molecule that regulates regeneration of injured neurons. By making adenoviral vector that transfers Shh, we found that the motor neuron survival was significantly promoted for 3-5 days after neonate rat injury. Furthermore, the continuous application of cyclopamin, which is a signal inhibitor of Smoothened, to the injured site, motor neuron death was induced in the adult rats after axotomy. These results indicate that Shh may play a regulatory role in the regeneration after nerve injury.

Asunto(s)

Proteínas Hedgehog/análisis , Regeneración Nerviosa/fisiología , Nervios Periféricos/química , Animales , Nervio Facial/química , Proteínas Hedgehog/fisiología , Ratas , Degeneración Walleriana/fisiopatología

RESUMEN

We report that the alternatively spliced isoforms of nonmuscle myosin heavy chain II-B (NHMC II-B) play distinct roles during mouse brain development. The B1-inserted isoform of NMHC II-B, which contains an insert of 10 amino acids near the ATP-binding region (loop 1) of the myosin heavy chain, is involved in normal migration of facial neurons. In contrast, the B2-inserted isoform, which contains an insert of 21 amino acids near the actin-binding region (loop 2), is important for postnatal development of cerebellar Purkinje cells. Deletion of the B1 alternative exon, together with reduced expression of myosin II-B, results in abnormal migration and consequent protrusion of facial neurons into the fourth ventricle. This protrusion is associated with the development of hydrocephalus. Restoring the amount of myosin II-B expression to wild-type levels prevents these defects, showing the importance of total myosin activity in facial neuron migration. In contrast, deletion of the B2 alternative exon results in abnormal development of cerebellar Purkinje cells. Cells lacking the B2-inserted isoform show reduced numbers of dendritic spines and branches. Some of the B2-ablated Purkinje cells are misplaced in the cerebellar molecular layer. All of the B2-ablated mice demonstrated impaired motor coordination.

Asunto(s)

Empalme Alternativo , Encéfalo/crecimiento & desarrollo , Hidrocefalia/genética , Miosina Tipo IIB no Muscular/fisiología , Animales , Encéfalo/anomalías , Encéfalo/patología , Movimiento Celular/genética , Extensiones de la Superficie Celular/genética , Extensiones de la Superficie Celular/patología , Cerebelo/anomalías , Cerebelo/crecimiento & desarrollo , Cerebelo/patología , Exones/genética , Nervio Facial/química , Nervio Facial/patología , Nervio Facial/fisiología , Hidrocefalia/patología , Ratones , Ratones Mutantes , Neuronas/química , Neuronas/patología , Neuronas/fisiología , Miosina Tipo IIB no Muscular/análisis , Miosina Tipo IIB no Muscular/genética , Isoformas de Proteínas , Células de Purkinje/química , Células de Purkinje/patología , Células de Purkinje/fisiología , Eliminación de Secuencia , Distribución Tisular , Transcripción Genética

RESUMEN

Presentation of antigen is key to the development of the immune response, mediated by association of antigen with major histocompatibility complex glycoproteins abbreviated as MHC1 and MHC2. In the current study, we examined the regulation of MHC1 in the brain after facial axotomy. The normal facial motor nucleus showed no immunoreactivity for MHC1 (MHC1-IR). Transection of the facial nerve led to a strong and selective up-regulation of MHC1-IR on the microglia in the affected nucleus, beginning at day 2 and reaching a maximum 14 days after axotomy, coinciding with a peak influx of the T lymphocytes that express CD8, the lymphocyte coreceptor for MHC1. Specificity of the MHC1 staining was confirmed in beta2-microglobulin-deficient mice, which lack normal cell surface MHC1-IR. MHC1-IR was particularly strong on phagocytic microglia, induced by delayed neuronal cell death, and correlated with the induction of mRNA for tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and interferon-gamma and the influx of T lymphocytes. Mice with severe combined immunodeficiency (scid), lacking T and B cells, showed an increase in the number of MHC1-positive nodules but no significant effect on overall MHC1-IR. Transgenic deletion of the IL1 receptor type I, or the interferon-gamma receptor type 1 subunit, did not affect the microglial MHC1-IR. However, a combined deletion of TNF receptors 1 and 2 (TNFR1&2-KO) led to a decrease in microglial MHC1-IR and to a striking absence of the phagocytic microglial nodules. Deletion of TNFR2 (p75) did not have an effect; deletion of TNFR1 (p55) reduced the diffuse microglial staining for MHC1-IR but did not abolish the MHC1(+) microglial nodules. In summary, neural injury leads to the induction of MHC1-IR on the activated, phagocytic microglia. This induction of MHC1 precedes the interaction with the immune system, at least in the facial motor nucleus model. Finally, the impaired induction of these molecules, up to now, only in the TNFR-deficient mice underscores the central role of TNF in the immune activation of the injured nervous system.

Asunto(s)

Antígenos CD/fisiología , Nervio Facial/fisiología , Glicoproteínas/biosíntesis , Antígenos de Histocompatibilidad Clase I/biosíntesis , Microglía/fisiología , Receptores del Factor de Necrosis Tumoral/fisiología , Animales , Antígenos CD/genética , Axotomía , Nervio Facial/química , Glicoproteínas/antagonistas & inhibidores , Antígenos de Histocompatibilidad Clase I/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones SCID , Microglía/química , Receptores del Factor de Necrosis Tumoral/deficiencia , Receptores del Factor de Necrosis Tumoral/genética , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral

RESUMEN

The common neurotrophin receptor (p75NTR) is a member of the tumor necrosis factor receptor superfamily and binds the neurotrophins nerve growth factor, brain derived neurotrophic factor, neurotrophin-3, and neurotrophin-4. P75NTR is expressed on developing motoneurons and is reexpressed on adult motoneurons under pathological conditions such as nerve trauma or neurodegeneration. Here we examined the regulation and function of p75NTR during regeneration after peripheral transection of the facial nerve of adult mice. Axotomy led to a strong increase in p75NTR immunoreactivity on the injured and regenerating facial motoneurons and on denervated Schwann cells. Cellular colocalization also revealed p75NTR immunoreactivity on neighboring blood vessels and cells in the injured nerve, but not on activated GFAP+ astrocytes or alphaMbeta2+ microglia and macrophages. To determine the function of this receptor we examined the effects of p75NTR deficiency on neuroglial activation, on the speed of axonal regeneration, and on neuronal survival after facial axotomy in two different transgenic mouse lines carrying targeted insertions exon 4 (p75e4-/-) or exon 3 (p75e3-/-) of the p75NTR gene. In both animal models absence of p75NTR led to a twofold, early increase in the number of CD3+. T-cells and in the microglial immunoreactivity for the alpha5beta1, alpha6beta1, and alphaMbeta2 integrins at day 4 in the facial nucleus and in the crushed facial motor nerve. No changes were observed in the number of reactive GFAP+ astrocytes or on late microglial and lymphocyte responses. The rate of axonal elongation in the crushed facial nerve, as well as neuronal survival, was found to be unaffected. Overall, the current study shows that the p75NTR receptor plays an important regulatory role in early neuroglial and immune activation.

Asunto(s)

Nervio Facial/fisiología , Neuronas Motoras/fisiología , Regeneración Nerviosa/fisiología , Receptores de Factor de Crecimiento Nervioso/fisiología , Animales , Nervio Facial/química , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuronas Motoras/química , Regeneración Nerviosa/genética , Ratas , Receptor de Factor de Crecimiento Nervioso , Receptores de Factor de Crecimiento Nervioso/análisis , Receptores de Factor de Crecimiento Nervioso/deficiencia , Receptores de Factor de Crecimiento Nervioso/genética

RESUMEN

OBJECTIVE: To explore the protein components and the biological neutrional activity upon motor neuron, so that take a general, basic study on the nerve regeneration microenvironment which be important to facial nerve regeneration. METHOD: The animal model of rabbit facial regeneration microenvironment is established. Advanced technology of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to study on the structural proteins in the regeneration fluid, as well as the motor neuron culture was used to test the biological activity in it. RESULT: 850 +/- 78 protein spots were detected in the regeneration fluid. The body area, neurite length and OD value of experimental group are higher than that of control group. CONCLUSION: We got the protein image of facial nerve regeneration fluid by 2-D PAGE, and proved that many kinds and number of proteins in the regeneration fluid have neutrional activity on motor neuron.

Asunto(s)

Nervio Facial/fisiología , Regeneración Nerviosa/fisiología , Animales , Electroforesis en Gel Bidimensional , Nervio Facial/química , Técnicas In Vitro , Neuronas Motoras/química , Neuronas Motoras/fisiología , Proteínas/análisis , Proteómica , Conejos

RESUMEN

We investigated whether the expression in the vestibular and facial nuclei of the voltage-dependent Na alpha I and Na alpha III channels and of the Ca(2+)-activated K(+)-channel subunits, small-conductance (SK) 1, SK2 and SK3, is affected by unilateral inner-ear lesion including both labyrinthectomy and transsection of the facial nerve. Specific sodium (Na alpha I, Na alpha III) and potassium (SK1, SK2, SK3) radioactive oligonucleotides were used to probe sections of rat vestibular and facial nuclei by in situ hybridization methods. The signal was detected with films or by emulsion photography. Animals were killed at various times following the lesion: 1 day, 3 days, 8 days or 30 days. In normal adult animals, mRNAs for Na alpha I, and SK1, SK2, and SK3 channels were found in several brainstem regions including the lateral, medial, superior and inferior vestibular nuclei and the facial nuclei. In contrast, there was little Na alpha III subunit mRNA anywhere in the brainstem. Following unilateral inner ear lesion in rats, the medial vestibular nuclei were probed with Na alpha I, Na alpha III, SK1, SK2 and SK3 oligonucleotide probes: autoradiography indicated no difference between the two sides, at any of the times studied. Na alpha I and SK2 mRNAs were less abundant and Na alpha III, SK1 and SK3 mRNAs were more abundant in the axotomized facial nuclei motoneurons than in controls. Removal of vestibular input did not affect the abundance of the mRNAs for the sodium- or calcium-dependent potassium channels in the deafferented vestibular nuclei. There is thus no evidence that modulation of these conductances contributes to the recovery of a normal resting discharge of the deafferented vestibular neurons and consequently to the functional recovery of the postural and oculomotor deficits observed at the acute stage. However, facial axotomy induced a long-term modulation of both Na and SK conductances mRNAs in the facial motoneurons ipsilateral to the lesion. Presumably, retrograde injury factors resulting from axotomy were able to alter durably the membrane properties and thus the excitability of the facial motoneurons.

Asunto(s)

Traumatismos del Nervio Facial , Nervio Facial/química , Canales de Potasio Calcio-Activados/análisis , Canales de Potasio con Entrada de Voltaje/análisis , Núcleos Vestibulares/química , Animales , Oído Interno/química , Oído Interno/fisiología , Nervio Facial/fisiología , Hibridación in Situ/métodos , Masculino , Canales de Potasio Calcio-Activados/fisiología , Canales de Potasio con Entrada de Voltaje/fisiología , Ratas , Ratas Long-Evans , ATPasa Intercambiadora de Sodio-Potasio/análisis , ATPasa Intercambiadora de Sodio-Potasio/fisiología , Núcleos Vestibulares/fisiología

RESUMEN

Cranial nerve impairment is one of the more serious complications in neurosurgery. Nevertheless, the important question of how many neurons are required for cranial nerve functions remains unanswered. The VIIth cranial nerve (facial nerve) in mice was subjected to graded crush injuries. After assessment of the facial function, the number of uninjured, healthy facial motor neurons was quantified with a retrograde neuronal tracer. We report that normal facial function is preserved if intact neurons account for more than 56% of the control value, while complete facial paralysis occurs if intact neurons are reduced to less than 32% of the control value.

Asunto(s)

Traumatismos del Nervio Facial , Nervio Facial/fisiología , Neuronas/fisiología , Animales , Nervio Facial/química , Masculino , Ratones , Neuronas/química

RESUMEN

There is accumulating evidence that cytokines are involved in the functioning of the brain and the spinal cord. However, it has been controversial whether they exert a neurotoxic or a neuroprotective effect. To address this question in vivo, we have examined the survival of injured motoneurons in a line of transgenic mice that overexpress the soluble form of tumour necrosis factor receptor-1 (sTNFR1). In these animals, all of the circulating TNF and lymphotoxin-alpha are neutralized by the continuous expression of the soluble receptor. Following axotomy of the facial nerve in 7-day-old control mice, we observed a loss of approximately 90% of the motoneurons at two weeks survival. In the transgenic mice under the same conditions, the percentage of motoneuron survival was increased two-fold (515 vs. 224) and varied as a function of the level of the circulating receptor. These results indicate that neutralization of endogenous TNF and lymphotoxin-alpha by means of overexpression of the soluble receptor can decrease cell death of injured motoneurons and suggest that these cytokines may play an important role in neuronal degeneration in the CNS following a lesion.

Asunto(s)

Neuronas Motoras/citología , Neuronas Motoras/fisiología , Receptores del Factor de Necrosis Tumoral/fisiología , Médula Espinal/citología , Factores de Edad , Animales , Axotomía , Supervivencia Celular/fisiología , Nervio Facial/química , Nervio Facial/fisiología , Linfotoxina-alfa/análisis , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Degeneración Nerviosa/fisiopatología , Solubilidad , Transgenes/fisiología , Factor de Necrosis Tumoral alfa/análisis

RESUMEN

I(2)-imidazoline receptors are mainly expressed on glial cells in the rat brain. This study was designed to test the effect of treatment with the I(2)-imidazoline selective receptor ligand LSL 60101 [2-(2-benzofuranyl)imidazole] on the morphology of astrocytes in the neonate and adult rat brain, and to explore the putative neuroprotective effects of this glial response. Short-term (3 days) or chronic (7-10 days) treatment with LSL 60101 (1 mg kg(-1), i.p. every 12 h) enhanced the area covered by astroglial cells in sections of facial motor nucleus from neonate rats processed for glial fibrillary acidic protein (GFAP) immunostaining. Facial motoneurons surrounded by positive glial cell processes were frequently observed in sections of LSL 60101-treated rats. A similar glial response was observed in the parietal cortex of adult rats after chronic (10 days) treatment with LSL 60101 (10 mg kg(-1), i.p. every 12 h). Western-blot detection of the specific astroglial glutamate transporter GLT-1, indicated increased immunoreactivity after LSL 60101 treatment in the pons of neonate and in the parietoccipital cortex of adult rats. In the facial motor nucleus of neonate rats, the glial response after LSL 60101 treatment was associated to a redistribution of the immunofluorescence of the basic fibroblast growth factor (FGF-2) from the perinuclear area of motoneurons to cover most of their cytoplasm, suggesting a translocation of this mitogenic and neurotrophic factor towards secretion pathways. The neuroprotective potential of the above effects of LSL 60101 treatment was tested after neonatal axotomy of facial motor nucleus. Treatment with LSL 60101 (1 mg kg(-1), i.p. every 12 h from day 0 to day 10 after birth) significantly reduced (38%) motoneuron death rate 7 days after facial nerve axotomy performed on day 3 after birth. It is concluded that treatment with the I(2)-imidazoline selective receptor ligand LSL 60101 provokes morphological/biochemical changes in astroglia that are neuroprotective after neonatal axotomy.

Asunto(s)

Astrocitos/efectos de los fármacos , Benzofuranos/farmacología , Muerte Celular/efectos de los fármacos , Imidazoles/farmacología , Neuronas Motoras/efectos de los fármacos , Receptores de Droga/agonistas , Transportadoras de Casetes de Unión a ATP/análisis , Sistema de Transporte de Aminoácidos X-AG , Animales , Animales Recién Nacidos , Astrocitos/citología , Western Blotting , Nervio Facial/química , Nervio Facial/citología , Nervio Facial/efectos de los fármacos , Factor 2 de Crecimiento de Fibroblastos/análisis , Proteína Ácida Fibrilar de la Glía/análisis , Receptores de Imidazolina , Inmunohistoquímica , Ligandos , Masculino , Neuronas Motoras/química , Neuronas Motoras/citología , Lóbulo Parietal/química , Lóbulo Parietal/citología , Lóbulo Parietal/efectos de los fármacos , Ratas , Ratas Sprague-Dawley

RESUMEN

Although expression of the small heat shock protein family member Hsp25 has been previously observed in the central nervous system (CNS), both constitutively and upon induction, its function in the CNS remains far from clear. In the present study we have characterized the spatial pattern of expression of Hsp25 in the normal adult mouse brain as well as the changes in expression patterns induced by subjecting mice to experimental hyperthermia or hypoxia. Immunohistochemical analysis revealed a surprisingly restricted pattern of constitutive expression of Hsp25 in the brain, limited to the facial, trigeminal, ambiguus, hypoglossal and vagal motor nuclei of the brainstem. After hyperthermia or hypoxia treatment, significant increases in the levels of Hsp25 were observed in these same areas and also in fibers of the facial and trigeminal nerve tracts. Immunoblot analysis of protein lysates from brainstem also showed the same pattern of induction of Hsp25. Surprisingly, no other area in the brain showed expression of Hsp25, in either control or stressed animals. The highly restricted expression of Hsp25 implies that this protein may have a specific physiological role in the orofacial motor nuclei, which govern precise coordination between muscles of mastication and the pharynx, larynx, and face. Its rapid induction after stress further suggests that Hsp25 may serve as a specific molecular chaperone in the lower cholinergic motor neurons and along their fibers under conditions of stress or injury.

Asunto(s)

Nervio Facial/citología , Proteínas de Choque Térmico , Neuronas Motoras/química , Proteínas de Neoplasias/fisiología , Estrés Fisiológico/fisiopatología , Núcleos del Trigémino/citología , Animales , Transporte Axonal/fisiología , Hipoxia de la Célula/fisiología , Músculos Faciales/inervación , Nervio Facial/química , Nervio Hipogloso/citología , Immunoblotting , Masculino , Ratones , Ratones Endogámicos ICR , Chaperonas Moleculares , Proteínas de Neoplasias/análisis

RESUMEN

Microglia represent a population of brain macrophage precursor cells which are intrinsic to the CNS parenchyma. Transection of the facial nerve in the newborn rat causes death of the affected motor neurons which is accompanied by massive activation of local microglia. Many of these cells develop into macrophages as can be shown by immunocytochemistry for OX-42 and ED1. Using the new polyclonal microglial marker ionized calcium binding adapter molecule 1, iba1, in combination with immunocytochemical double-labeling for the proliferating cell nuclear antigen (PCNA), or [3H]thymidine autoradiography, and confocal microscopy, qualitative as well as quantitative differences can be demonstrated between the newborn and the adult axotomized rat facial nucleus. While microglial cells are the only cell population which responds to axotomy by cell division in the adult facial nucleus, GFAP positive reactive astrocytes can be shown to undergo mitosis following axotomy in the newborn rat. Furthermore, ED1 immunoreactivity, early expression of MHC class II molecules and morphological transformation of microglia into macrophages can only be observed under conditions of neuronal degeneration, i.e., in the neonatal rat facial nucleus. Thus, the combination of cellular markers described here should be useful for studies employing the neonatal rat facial nucleus as an in vivo assay system to test the efficacy of neurotrophic factors.

Asunto(s)

Nervio Facial/citología , Macrófagos/inmunología , Microglía/inmunología , Factores de Edad , Animales , Animales Recién Nacidos , Axotomía , Proteínas de Unión al Calcio/análisis , División Celular/inmunología , Nervio Facial/química , Nervio Facial/cirugía , Complejo Mayor de Histocompatibilidad , Masculino , Proteínas de Microfilamentos , Neuronas Motoras/química , Degeneración Nerviosa/inmunología , Antígeno Nuclear de Célula en Proliferación/análisis , Ratas , Ratas Wistar

RESUMEN

Voltage-dependent Ca2+ channels are a major pathway for Ca2+ entry in neurons. We have studied the electrophysiological, pharmacological, and molecular properties of voltage-gated Ca2+ channels in motoneurons of the rat facial nucleus in slices of the brainstem. Most facial motoneurons express both low voltage-activated (LVA) and high voltage-activated (HVA) Ca2+ channel currents. The HVA current is composed of a number of pharmacologically separable components, including 30% of N-type and approximately 5% of L-type. Despite the dominating role of P-type Ca2+ channels in transmitter release at facial motoneuron terminals described in previous studies, these channels were not present in the cell body. Remarkably, most of the HVA current was carried through a new type of Ca2+ channel that is resistant to toxin and dihydropyridine block but distinct from the R-type currents described in other neurons. Using reverse transcription followed by PCR amplification (RT-PCR) with a powerful set of primers designed to amplify all HVA subtypes of the alpha1-subunit, we identified a highly heterogeneous expression pattern of Ca2+ channel alpha1-subunit mRNA in individual neurons consistent with the Ca2+ current components found in the cell bodies and axon terminals. We detected mRNA for alpha1A in 86% of neurons, alpha1B in 59%, alpha1C in 18%, alpha1D in 18%, and alpha1E in 59%. Either alpha1A or alpha1B mRNAs (or both) were present in all neurons, together with various other alpha1-subunit mRNAs. The most frequently occurring combination was alpha1A with alpha1B and alpha1E. Taken together, these results demonstrate that the Ca2+ channel pattern found in facial motoneurons is highly distinct from that found in other brainstem motoneurons.

Asunto(s)

Canales de Calcio/genética , Nervio Facial/citología , Neuronas Motoras/química , Receptores Colinérgicos/genética , omega-Conotoxinas , Animales , Animales Recién Nacidos , Tronco Encefálico/citología , Cadmio/farmacología , Bloqueadores de los Canales de Calcio/farmacología , Canales de Calcio Tipo L , ADN Complementario , Dihidropiridinas/farmacología , Nervio Facial/química , Activación del Canal Iónico/efectos de los fármacos , Níquel/farmacología , Nitrendipino/farmacología , Técnicas de Placa-Clamp , Péptidos/farmacología , ARN Mensajero/análisis , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Venenos de Araña/farmacología , omega-Agatoxina IVA , omega-Conotoxina GVIA

RESUMEN

Sensory neurons of the olfactory, trigeminal, facial, vestibulo-cochlear, glossopharyngeal and vagal nerves, and neurons migrating along the olfactory nerve to the brain have special anlagen, made up of placodes located in the epithelial layer. To investigate the characteristic phenotype of placode-derived neurons, immunohistochemical analysis of intermediate filaments was conducted on formalin-fixed human embryonic tissues. Neurons arising from placodes including luteinizing-hormone releasing hormone (LHRH) neurons migrating from the olfactory placode to the brain had immunoreactivity to antibodies specific to cytokeratin, AE1 and CAM5.2 during the embryonic stage. However, this immunoreactivity disappeared during the late embryonic to the post-embryonic stage and was not observed in the roots of these nerves in the post-natal stage. Immunoreactivity was detected in both the somata and processes, and the distribution differed from that described in rodent brain neurons. With this exception, no other human peripheral neurons, including spinal dorsal root ganglia, had immunoreactivity with anti-cytokeratin antibodies throughout the entire developmental stage. Although the cephalic neural crest also directly generates neurons to most of the cranial sensory ganglia, we could not find any evidence that it contributed to the genesis of cytokeratin-positive embryonic neurons. We concluded that cytokeratin is an intermediate filament common to human embryonic neurons of cephalic placodal origin and that this immunohistochemical marker may be useful in analyzing the developmental sequence of several congenital diseases involving the cranial nerves, such as Moebius syndrome and Goldenhar syndrome.

Asunto(s)

Queratinas/análisis , Queratinas/inmunología , Cresta Neural/citología , Neuronas Aferentes/química , Adulto , Especificidad de Anticuerpos , Diferenciación Celular/fisiología , Nervios Craneales/química , Nervios Craneales/citología , Nervios Craneales/embriología , Nervio Facial/química , Nervio Facial/citología , Nervio Facial/embriología , Humanos , Inmunohistoquímica , Recién Nacido , Mesodermo/química , Mesodermo/citología , Cresta Neural/química , Cresta Neural/embriología , Ganglio Nudoso/química , Ganglio Nudoso/citología , Ganglio Nudoso/embriología , Nervio Olfatorio/química , Nervio Olfatorio/citología , Nervio Olfatorio/embriología , Médula Espinal/química , Médula Espinal/citología , Médula Espinal/embriología , Nervio Trigémino/química , Nervio Trigémino/citología , Nervio Trigémino/embriología , Nervio Vestibulococlear/química , Nervio Vestibulococlear/citología , Nervio Vestibulococlear/embriología

RESUMEN

The transient expression of estrogen receptor (ER) in the ventromedial subnucleus of the facial nucleus was previously detected in the newborn rat, and the expression of ER molecules was down-regulated by daily injections of estradiol. Here we examined possible involvement of aromatization in this process. ER molecules were measured by immunohistochemistry and in situ hybridization histochemistry after daily injections of testosterone propionate (TP; 100 micrograms/0.02 ml) and estradiol benzoate (EB; 10 micrograms/0.02 ml) in the male pups castrated within 24 h of birth. Daily injections of TP for 5 consecutive days did not suppress ER and ER mRNA in the facial nucleus, while they were both suppressed by daily injections of EB. Moreover, aromatase immunoreactivity was not detected in the facial nucleus of both castrated, TP injected and intact control males at 6 days of age. The present findings therefore suggest that ER molecules expressed transiently in the facial nucleus are not directly involved in masculine sexual differentiation of the brain in newborn rat.

Asunto(s)

Aromatasa/metabolismo , Estradiol/farmacología , Bulbo Raquídeo/ultraestructura , Receptores de Estrógenos/biosíntesis , Testosterona/farmacología , Animales , Animales Recién Nacidos , Nervio Facial/química , Nervio Facial/citología , Nervio Facial/enzimología , Inmunohistoquímica , Hibridación in Situ , Masculino , Bulbo Raquídeo/química , Bulbo Raquídeo/enzimología , ARN Mensajero/análisis , Ratas , Ratas Sprague-Dawley , Receptores de Estrógenos/genética

RESUMEN

Mammalian lateral olivocochlear (LOC) neurons that are immunoreactive for choline acetyltransferase (ChAT) are also immunoreactive for enkephalin (Enk). To determine whether cochlear efferent neurons in birds might also contain Enk-like immunoreactivity (Enk-LI), we studied the auditory brainstem of the domestic chicken using antisera to ChAT, leucine-enkephalin (L-Enk) and methionine-enkephalin (M-Enk). Enk-LI terminals are found around, but not within, the superior olivary nucleus (SO) and the nucleus of the lateral lemniscus, pars intermedia (LLi). A moderate concentration of Enk-LI terminals is found ventromedial to the ventral facial nucleus (VIIv) where the ventrolateral group of ChAT-I cochlear efferent neurons is located. After colchicine injections into the lateral ventricle, a population of intensely stained Enk-LI perikarya was found in the nucleus of the lateral lemniscus, pars ventralis (LLv) with scattered cells in the LLi and the nucleus subceruleus ventralis (SCv). The distribution of Enk-LI and ChAT-I somata, however, never overlapped, even in adjacent sections. Thus, in the chick, Enk-LI perikarya are not distributed in areas where cochlear efferent neurons are found. Instead, a dense concentration of Enk-I terminals can be found in areas containing ChAT-I cochlear efferent neurons. The source of these enkephalinergic terminals may be a population of Enk-LI cells in the LLv.

Asunto(s)

Tronco Encefálico/química , Pollos/anatomía & histología , Cóclea/citología , Encefalinas/análisis , Neuronas Eferentes/química , Animales , Tronco Encefálico/citología , Tamaño de la Célula/efectos de los fármacos , Tamaño de la Célula/fisiología , Colina O-Acetiltransferasa/análisis , Cóclea/efectos de los fármacos , Colchicina/administración & dosificación , Colchicina/farmacología , Encefalina Leucina/análisis , Nervio Facial/química , Nervio Facial/citología , Inmunohistoquímica , Neuronas Eferentes/citología , Núcleo Olivar/química , Núcleo Olivar/efectos de los fármacos , Núcleos del Trigémino/química , Núcleos del Trigémino/citología , Nervio Vago/química , Nervio Vago/citología

RESUMEN

This paper presents the morphologic charactistic of substance P immunoreactivity (SP-IR) in the facial nerve and geniculate ganglion of normal cats by using immunohistochemical technique and immunoelectronic microscopy. SP-IR positive cells are more in geniculate ganglion than the other parts of the facial nerve. These cells are black. The sizes of their somata are different, and the shapes are round, elliptic or irregular. The somata are connected with each other by their central and peripheral synapses in the shapes of spider's web or root of a tree. Among the cells the bipolar and pseudounipolar cells are also found. SP-IR positive fibers curve and some of them have obvious expansive bodies, which are beaded, different in diameter. Under electronic microscope SP-IR positive cells show thicker electronic density than the others, and SP-IR positive products with thick electronic density are found in the nuclei, nucleomembranes, outer membranes of mitochndria and plasma. The nerve fibers are myelinated and the myelin sheath is concentric, and in the axoplasms there are nervous microfilament and a bit of mitochndria.

Asunto(s)

Nervio Facial/química , Ganglio Geniculado/química , Sustancia P/metabolismo , Animales , Gatos , Nervio Facial/ultraestructura , Femenino , Ganglio Geniculado/ultraestructura , Inmunohistoquímica , Masculino , Microscopía Inmunoelectrónica

Asunto(s)

Tronco Encefálico/química , Péptido Relacionado con Gen de Calcitonina/análisis , Colina O-Acetiltransferasa/análisis , Nervio Facial/química , Anciano , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad

RESUMEN

This study characterizes the temporal-spatial distribution of nerve growth factor (NGF) low (p75) and high-affinity (trkA) receptors in the facial nerve and geniculate ganglion (GG) of developing quail embryos (E-3 to E-14). We used 125I-labeled NGF (125I-NGF) to study binding dynamics in a temporal series of isolated primordia and an autoradiographic series of staged specimens to characterize the occurrence and distribution of NGF receptors in this cranial nerve and its ganglion. In addition, expression of trkA and p75 protein-like immunoreactivity in the facial nerve and GG was studied by Western blot, in order to distinguish between high- and low-affinity NGF receptors respectively. The quantitative study of binding show that isolated facial primordia ranging from E-3 to E-14 exhibit different levels of specific binding. High initial binding levels were observed on E-3 specimens, then an initial decrease on day 4 (E-4) followed by a steady increase from days E-4 to E-7. Maximum 125I-NGF binding was achieved on E-7, followed by a steady decline in binding on days 8 (E-8) and 9 (E-9), reaching near background levels on day 10 (E-10) of development and until the oldest stage assayed (E-14). Most of the cells bearing NGF receptors appeared to be non-neuronal crest-derived cells, but some placode-derived neurons and motor fibers of the VIIth cranial nerve transiently expressed the ability to bind 125I-NGF. The temporal pattern of p75 expression matches the pattern of quantitative binding of NGF, while the trkA expression is restricted to a few stages mainly E7 and E9, implying that most of the binding detected is via low-affinity receptors, except for a proportion of high-affinity receptors present at stages of maximum binding. This temporal pattern of NGF binding sites suggests that cells within the VIIth cranial nerve are responsive to and/or dependent upon NGF in vivo, so NGF may play a biological role during normal development of the facial nerve. In view of the developmental events that parallel the occurrence and type of NGF binding sites, we suggest that this role may be to modulate from earlier chemotaxis and cell proliferation to much later events, such as neuronal differentiation and neuron-glia interactions. The significance of these findings in regeneration during adult life remain to be investigated.

Asunto(s)

Coturnix/embriología , Nervio Facial/química , Glicoproteínas de Membrana/análisis , Glicoproteínas de Membrana/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Proteínas Proto-Oncogénicas/análisis , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Tirosina Quinasas Receptoras/análisis , Proteínas Tirosina Quinasas Receptoras/metabolismo , Receptores de Factor de Crecimiento Nervioso/análisis , Receptores de Factor de Crecimiento Nervioso/metabolismo , Animales , Autorradiografía , Western Blotting , Nervio Facial/metabolismo , Nervio Facial/ultraestructura , Ganglio Geniculado/química , Ganglio Geniculado/metabolismo , Ganglio Geniculado/ultraestructura , Radioisótopos de Yodo , Factores de Crecimiento Nervioso/análisis , Receptor de Factor de Crecimiento Nervioso , Receptor trkA

RESUMEN

Facial nerve transection leads to functional and structural reactions in lesioned motor neurones and surrounding glial cells. Data from this study provide evidence that the most rapid reaction described so far consists of an increase in immunoreactivity of connexin-43 (cx-43), the predominant gap junction protein in astrocytes. The ipsilateral facial nucleus is selectively marked as early as 0.75 to 1.5 hours after axotomy, while the unlesioned side as well as the unoperated controls remain faintly stained. Thus, enhanced coupling capacity of astrocytes by gap junctions appears to be a sensitive indicator of modified neuronal-glial interaction in the CNS.

Asunto(s)

Astrocitos/citología , Axones/fisiología , Conexina 43/análisis , Nervio Facial/fisiología , Animales , Comunicación Celular/fisiología , Nervio Facial/química , Neuronas Motoras/citología , Neuroglía/citología , Ratas , Ratas Sprague-Dawley , Factores de Tiempo