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首页> 外文期刊>The European Journal of Neuroscience >Neuronal differentiation of the early embryonic auditory hindbrain of the chicken in primary culture.
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Neuronal differentiation of the early embryonic auditory hindbrain of the chicken in primary culture.

机译：鸡的早期胚胎听觉后脑在原代培养中的神经元分化。

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Neurons in the auditory hindbrain pathway of the chicken are physiologically and morphologically highly specialized. It remains unclear to what extent independent differentiation vs. activity-dependent mechanisms determines the development of this system. To address this question we established a primary culture system of the early auditory hindbrain neurons. Primary cultures of neurons from nucleus magnocellularis and nucleus laminaris were prepared from embryonic day 6.5 chicken. These cells developed in culture under serum-free conditions for up to 15 days. Immunocytochemical staining and whole-cell patch recordings were used to characterize the development of the neurons. A stable expression of the calcium-binding protein calretinin, which serves as a characteristic marker of the auditory pathway, was found at all stages. A voltage-gated potassium channel (Kv3.1b) with a specific function in the auditory system was also expressed after about 1 week in culture. Electrophysiological recordings showeda general maturation of the neuronal phenotype as reflected by an increase in the mean resting membrane potential, a decrease in the mean input resistance as well as a maturation of action potential parameters. Four groups of neurons that generate action potentials could be distinguished. One of these showed the phasic firing pattern of auditory brainstem neurons known from slice preparations. In older cultures we demonstrated functional synaptogenesis in vitro by recording postsynaptic activity elicited by extracellular stimulation and styryl dye loading of vesicles. Thus, isolated neurons from the auditory region of the avian brainstem differentiate to specific neuronal subtypes and autonomously develop synaptic connections in vitro.
机译：鸡的听觉后脑途径中的神经元在生理和形态上高度专门化。目前尚不清楚独立分化与活动依赖机制在多大程度上决定了该系统的发展。为了解决这个问题，我们建立了早期听觉后脑神经元的主要培养系统。从第6.5天的胚胎鸡中制备了来自巨细胞核和层状核的神经元的原代培养物。这些细胞在无血清条件下培养长达15天。免疫细胞化学染色和全细胞膜片记录被用来表征神经元的发育。在所有阶段都发现了钙结合蛋白钙调蛋白的稳定表达，该蛋白可以作为听觉途径的特征标记。培养约1周后，还表达了在听觉系统中具有特定功能的电压门控钾通道（Kv3.1b）。电生理学记录显示神经元表型的一般成熟，反映为平均静息膜电位增加，平均输入阻力减少以及动作电位参数成熟。可以区分出产生动作电位的四组神经元。其中之一显示了切片制备中已知的听性脑干神经元的阶段性放电模式。在较老的文化中，我们通过记录细胞外刺激和囊泡的苯乙烯基染料负载引发的突触后活性，证明了体外的功能性突触形成。因此，从禽脑干听觉区域分离的神经元分化为特定的神经元亚型，并在体外自主发展突触连接。

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