Corrigall W A, Crain S M, Bornstein M B
J Neurobiol. 1976 Nov;7(6):521-36. doi: 10.1002/neu.480070606.
Explants of 18- or 19-day fetal mouse olfactory bulb have been maintained in culture for periods up to 5 weeks. Compound action potentials can be evoked in the bulb explants by 1 day in vitro, and by 3-4 days, synaptically mediated slow wave discharges can be domonstrated in bicuculline (10(-5) m). The capability of the bulb explants to generate these slow-wave discharges has also been revealed by the introduction of picrotoxin (10(-5)m, d-tubocurarine (10(-4)m) and chloride-free medium, but not of strychnine (up to 3 X 10(-5)M. The data indicate early functional development of inhibitory, as well as excitatory, synaptic systems. In addition, the selective and reversible depression of these slow wave potentials by GABA (1-5 X 10(-4)M), but not by glycine (up to 3 X 10(-3)M), indicates a GABA-ergic component in the inhibitory network. Single unit extracellular recordings have been obtained from the presumptive mitral cells which, in culture, are spontaneously active even as early as 1-2 days after explantation. Correlative Bodian silver-impregnations demonstrate the presence of neurons in these explants which resemble typical mitral cells. Studies of mitral cells using paired stimuli suggest the development in vitro of an inhibitory system analogous to that known to suppress the excitability of their in situ counterparts following orthodromic or antidromic activation. These data, as well as the pharmacological sensitivities of the mitral cells in culture to GABA (5 X 10(-4)M) and bicuculline (10(-5)M), indicate that granule-to-mitral synapses may develop characteristic functions in olfactory bulb explants.
18或19日龄胎鼠嗅球外植体已在培养中维持长达5周的时间。体外培养1天时,嗅球外植体就能诱发复合动作电位,到3至4天时,在荷包牡丹碱(10⁻⁵M)存在的情况下,可证明有突触介导的慢波放电。引入印防己毒素(10⁻⁵M)、d -筒箭毒碱(10⁻⁴M)和无氯培养基也能揭示嗅球外植体产生这些慢波放电的能力,但士的宁(高达3×10⁻⁵M)则不能。数据表明抑制性和兴奋性突触系统早期就有功能发育。此外,γ-氨基丁酸(1 - 5×10⁻⁴M)能选择性且可逆地抑制这些慢波电位,而甘氨酸(高达3×10⁻³M)则不能,这表明抑制性网络中存在γ-氨基丁酸能成分。已从假定的僧帽细胞进行了单细胞胞外记录,在培养中,这些细胞甚至在植入后1至2天就自发活跃。相关的博迪安银浸染显示这些外植体中存在类似于典型僧帽细胞的神经元。使用成对刺激对僧帽细胞的研究表明,体外发育出了一种抑制系统,类似于已知的在顺向或逆向激活后抑制其原位对应物兴奋性的系统。这些数据以及培养中的僧帽细胞对γ-氨基丁酸(5×10⁻⁴M)和荷包牡丹碱(10⁻⁵M)的药理学敏感性表明,颗粒细胞到僧帽细胞的突触可能在嗅球外植体中发展出特征性功能。