RESUMO
A crucial step in the development of the vertebrate visual system is the branching of retinal ganglion cell (RGC) axons within their target, the superior colliculus/tectum. A major player in this process is the neurotrophin brain-derived neurotrophic factor (BDNF). However, the molecular basis for the signaling pathways mediating BDNF action is less well understood. As BDNF exerts some of its functions by controlling the expression of microRNAs (miRNAs), we investigated whether miRNAs are also involved in BDNF-mediated retinal axon branching. Here, we demonstrate that the expression pattern of miRNA-132 in the retina is consistent with its involvement in this process, and that BDNF induces the upregulation of miRNA-132 in retinal cultures. Furthermore, in vitro gain-of-function and loss-of-function approaches in retinal cultures reveal that miRNA-132 mediates axon branching downstream of BDNF. A known target of miRNA-132 is the Rho family GTPase-activating protein, p250GAP. We find that p250GAP is expressed in RGC axons and mediates the effects of miRNA-132 in BDNF-induced branching. BDNF treatment or overexpression of miRNA-132 leads to a reduction in p250GAP protein levels in retinal cultures, whereas the overexpression of p250GAP abolishes BDNF-induced branching. Finally, we used a loss-of-function approach to show that miRNA-132 affects the maturation of RGC termination zones in the mouse superior colliculus in vivo, while their topographic targeting remains intact. Together, our data indicate that BDNF promotes RGC axon branching during retinocollicular/tectal map formation via upregulation of miRNA-132, which in turn downregulates p250GAP.
Assuntos
Axônios/metabolismo , Fator Neurotrófico Derivado do Encéfalo/farmacologia , Proteínas Ativadoras de GTPase/fisiologia , MicroRNAs/fisiologia , Células Ganglionares da Retina/metabolismo , Animais , Axônios/efeitos dos fármacos , Células Cultivadas , Embrião de Galinha , Feminino , Proteínas Ativadoras de GTPase/deficiência , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Células Ganglionares da Retina/efeitos dos fármacosRESUMO
At the Drosophila neuromuscular junction (NMJ), the loss of retrograde, trans-synaptic BMP signaling causes motoneuron terminals to have fewer synaptic boutons, whereas increased neuronal activity results in a larger synapse with more boutons. Here, we show that an early and transient BMP signal is necessary and sufficient for NMJ growth as well as for activity-dependent synaptic plasticity. This early critical period was revealed by the temporally controlled suppression of Mad, the SMAD1 transcriptional regulator. Similar results were found by genetic rescue tests involving the BMP4/5/6 ligand Glass bottom boat (Gbb) in muscle, and alternatively the type II BMP receptor Wishful Thinking (Wit) in the motoneuron. These observations support a model where the muscle signals back to the innervating motoneuron's nucleus to activate presynaptic programs necessary for synaptic growth and activity-dependent plasticity. Molecular genetic gain- and loss-of-function studies show that genes involved in NMJ growth and plasticity, including the adenylyl cyclase Rutabaga, the Ig-CAM Fasciclin II, the transcription factor AP-1 (Fos/Jun), and the adhesion protein Neurexin, all depend critically on the canonical BMP pathway for their effects. By contrast, elevated expression of Lar, a receptor protein tyrosine phosphatase found to be necessary for activity-dependent plasticity, rescued the phenotypes associated with the loss of Mad signaling. We also find that synaptic structure and function develop using genetically separable, BMP-dependent mechanisms. Although synaptic growth depended on Lar and the early, transient BMP signal, the maturation of neurotransmitter release was independent of Lar and required later, ongoing BMP signaling.
Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Plasticidade Neuronal/fisiologia , Neurônios/metabolismo , Transdução de Sinais/fisiologia , Sinapses/metabolismo , Adenilil Ciclases/genética , Adenilil Ciclases/metabolismo , Animais , Animais Geneticamente Modificados , Proteínas Morfogenéticas Ósseas/genética , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Drosophila , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Neurônios Motores/metabolismo , Junção Neuromuscular/genética , Junção Neuromuscular/metabolismo , Proteínas Tirosina Fosfatases Semelhantes a Receptores/genética , Proteínas Tirosina Fosfatases Semelhantes a Receptores/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Sinapses/genética , Transmissão Sináptica/fisiologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
Recent advances in the study of axon guidance have begun to clarify the intricate signalling mechanisms utilised by receptors that mediate path-finding. Many of these axon guidance receptors, including Plexin B, EphA, ephrin B and Robo, regulate the Rho family of GTPases, to effect changes in motility. Recent studies demonstrate a critical role for the cytoplasmic tails of guidance receptors in signalling and also reveal the potential for a great deal of crosstalk between the various receptor-signalling pathways.