RESUMO
Peripheral nerve injury results in plastic changes in the dorsal root ganglia and spinal cord, and is often complicated with neuropathic pain. The mechanisms underlying these changes are not known. We have now investigated the expression of brain-derived neurotrophic factor in the dorsal root ganglia with histochemical and biochemical methods following sciatic nerve lesion in the rat. The percentage of neurons immunoreactive for brain-derived neurotrophic factor in the ipsilateral dorsal root ganglia was significantly increased as early as 24 h after the nerve lesion and the increase lasted for at least two weeks. The level of brain-derived neurotrophic factor messenger RNA was also significantly increased in the ipsibut not contralateral dorsal root ganglia. Both neurons and satellite cells in the lesioned dorsal root ganglia synthesized brain-derived neurotrophic factor messenger RNA after the nerve lesion. There was a dramatic shift in size distribution of positive neurons towards large sizes seven days after sciatic nerve lesion. Morphometric analysis and retrograde tracing studies showed that no injured neurons smaller than 600 microm2 were immunoreactive for brain-derived neurotrophic factor, whereas the majority of large injured neurons were immunoreactive in the ipsilateral dorsal root ganglia seven days postlesion. The brain-derived neurotrophic factor-immunoreactive nerve terminals in the ipsilateral spinal cord were reduced in the central region of lamina II, but increased in more medial regions or deeper into laminae III/IV. These studies indicate that sciatic nerve injury results in a differential regulation of brain-derived neurotrophic factor in different subpopulations of sensory neurons in the dorsal root ganglia. Small neurons switched off their normal synthesis of brain-derived neurotrophic factor, whereas larger ones switched to a brain-derived neurotrophic factor phenotype. The phenotypic switch may have functional implications in neuronal plasticity and generation of neuropathic pain after nerve injury.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Neurônios Aferentes/metabolismo , Nervo Isquiático/lesões , Ferimentos e Lesões/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Contagem de Células , Tamanho Celular , Feminino , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Masculino , Neurônios Aferentes/patologia , Neurônios Aferentes/fisiologia , Fenótipo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/patologia , Nervo Isquiático/fisiologia , Medula Espinal/metabolismo , Ferimentos e Lesões/genéticaRESUMO
In a previous study we have shown that a subpopulation of primary sensory neurons contain brain-derived neurotrophic factor immunoreactivity. In the present study we investigated the distribution of brain-derived neurotrophic factor and its mRNA in cranial and spinal ganglia at different segmental levels, using immunohistochemical and quantitative reverse transcriptase-polymerase chain reaction techniques. Our results show that there is no significant difference in the percentage of brain-derived neurotrophic factor-immunoreactive neurons in spinal ganglia of different segmental levels. In contrast, more brain-derived neurotrophic factor-immunoreactive neurons were found in placode-derived than neural crest-derived ganglia. The percentage of brain-derived neurotrophic factor-immunoreactive neurons is consistent with the percentage of neurons lost after deletion of brain-derived neurotrophic factor or trkB genes. However, there is no correlation between brain-derived neurotrophic factor mRNA levels and the number of brain-derived neurotrophic factor immunoreactive neurons in these ganglia, suggesting that some neurons synthesize brain-derived neurotrophic factor while others accumulate the factor following its retrograde transport within nerve fibers. In particular, the proportion of brain-derived neurotrophic factor that is derived from extraganglionic sources in the placode-derived ganglia appears greater than that in the neural crest-derived ganglia.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Nervos Cranianos/metabolismo , Gânglios Espinais/metabolismo , Gânglios/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Feminino , Gânglios/citologia , Gânglios Espinais/citologia , Imuno-Histoquímica , Masculino , Neurônios Aferentes/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
In a previous study we have demonstrated that endogenous neurotrophin 3 (NT3) is required for the survival of most sympathetic neurones in postnatal rats. However, the mechanisms underlying the action of NT3 on sympathetic neurones is not known. Neither is it understood whether NT3 is retrogradely transported from peripheral tissues or acts locally in an autocrine fashion. In the present study, NT3-mRNA was quantified in sympathetic effector tissues and NT3 protein was localized in intact and lesioned sympathetic nerves in rats. NT3-mRNA is expressed and developmentally regulated in many effector tissues including mesenteric arteries, salivary gland, heart and kidney but hardly detectable in the superior cervical ganglia of adult animals. The majority of sympathetic neurones express immunoreactivity for TrkA and TrkC in both neonatal and adult rats. Sympathetic somata are normally immunoreactive for NT3, but the immunoreactivity is abolished by systemic administration of NT3 antibodies or axotomy of postganglionic neurones, suggesting an accumulation of NT3 from extraneuronal sources. Furthermore, the detection of NT3-immunoreactivity in the internal carotid nerve as early as 3 h following a compression and only on the distal side indicates endogenous NT3 is retrogradely transported by sympathetic neurones. These studies provide evidence indicating that NT3, like nerve growth factor, is an effector tissue-derived neurotrophic factor for sympathetic neurones both during development and in the adult animal. Thus, we have provided a clear example that one type of neurone derives, through a retrograde transport mechanism, two neurotrophic factors simultaneously from its peripheral effector tissues.