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Neuronal differentiation of the early embryonic auditory hindbrain of the chicken in primary culture
Author(s) -
Kuenzel Thomas,
Mönig Benedikt,
Wagner Hermann,
Mey Jörg,
Luksch Harald
Publication year - 2007
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1111/j.1460-9568.2007.05343.x
Subject(s) - neuroscience , biology , hindbrain , synaptogenesis , electrophysiology , brainstem , premovement neuronal activity , afterhyperpolarization , auditory system , cellular neuroscience , superior olivary complex , cochlear nucleus , calretinin , microbiology and biotechnology , central nervous system , immunohistochemistry , immunology
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 showed a 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 .