Dipartmento di Morfologia, Biochimica, Fisiologia e Produzione Animale, Sezione di Morfologia, Università di Messina, Messina, Italy.
Cell Mol Neurobiol. 2010 Jul;30(5):787-93. doi: 10.1007/s10571-010-9506-z. Epub 2010 Feb 17.
The neuromasts of the lateral line system are regarded as a model to study the mechanisms of hearing, deafness, and ototoxicity. The neurotrophins (NTs), especially brain-derived neurotrophic factor (BDNF), and its signaling receptor TrkB are involved in the development and maintenance of neuromasts. To know the period in which the BDNF/TrkB complex has more effects in the neuromast biology, the age-related changes were studied. Normal zebrafish from 10 to 180 days post-fertilization (dpf), as well as transgenic ET4 zebrafish 10 and 20 dpf, was analyzed using qRT-PCR, western blot, and immunohistochemistry. BDNF and TrkB mRNAs followed a parallel course, peaking at 20 dpf, and thereafter progressively decreased. Specific immunoreactivity for BDNF and TrkB was found co-localized in all hairy cells of neuromasts in 20 and 30 dpf; then, the number of immunoreactive cells decreased, and by 180 dpf BDNF remains restricted to a subpopulation of hairy cells, and TrkB to a few number of sensory and non-sensory cells. At all ages examined, TrkB immunoreactivity was detected in sensory ganglia innervating the neuromasts. The present results demonstrate that there is a parallel time-related decline in the expression of BDNF and TrkB in zebrafish. Also, the patterns of cell expression suggest that autocrine/paracrine mechanisms for this NT system might occur within the neuromasts. Because TrkB in lateral line ganglia did not vary with age, their neurons are potentially capable to respond to BDNF during the entire lifespan of zebrafish.
侧线系统的神经丘被认为是研究听觉、耳聋和耳毒性机制的模型。神经营养因子(NTs),尤其是脑源性神经营养因子(BDNF)及其信号受体 TrkB,参与了神经丘的发育和维持。为了了解 BDNF/TrkB 复合物在神经丘生物学中具有更多影响的时期,研究了年龄相关的变化。使用 qRT-PCR、western blot 和免疫组织化学分析了从受精后 10 天到 180 天的正常斑马鱼以及 10 天和 20 天的 ET4 转基因斑马鱼。BDNF 和 TrkB mRNA 呈平行趋势,在 20 dpf 时达到峰值,此后逐渐下降。在 20 天和 30 天的神经丘中,BDNF 和 TrkB 的特异性免疫反应均与所有刚毛细胞共定位;然后,免疫反应细胞的数量减少,到 180 天,BDNF 仍然局限于刚毛细胞的亚群,而 TrkB 局限于少数感觉和非感觉细胞。在所有检查的年龄中,TrkB 免疫反应性都在支配神经丘的感觉神经节中被检测到。本研究结果表明,BDNF 和 TrkB 在斑马鱼中的表达呈平行的时间相关下降。此外,细胞表达模式表明,该 NT 系统的自分泌/旁分泌机制可能发生在神经丘内。由于侧线神经节中的 TrkB 不随年龄变化,它们的神经元在斑马鱼的整个生命周期中都有可能对 BDNF 做出反应。