Neuroscience Institute and Department of Biology, Georgia State University, P.O. Box 5030, Atlanta, GA, 30302-5030, USA.
J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2014 Jan;200(1):53-76. doi: 10.1007/s00359-013-0861-3. Epub 2013 Nov 1.
We determined if a newly developed antennule slice preparation allows studying chemosensory properties of spiny lobster olfactory receptor neurons under in situ conditions with Ca(2+) imaging. We show that chemical stimuli reach the dendrites of olfactory receptor neurons but not their somata, and that odorant-induced Ca(2+) signals in the somata are sufficiently stable over time to allow stimulation with a substantial number of odorants. Pharmacological manipulations served to elucidate the source of odorant-induced Ca(2+) transients and spontaneous Ca(2+) oscillations in the somata of olfactory receptor neurons. Both Ca(2+) signals are primarily mediated by an influx of extracellular Ca(2+) through voltage-activated Ca(2+) channels that can be blocked by CoCl2 and the L-type Ca(2+) channel blocker verapamil. Intracellular Ca(2+) stores contribute little to odorant-induced Ca(2+) transients and spontaneous Ca(2+) oscillations. The odorant-induced Ca(2+) transients as well as the spontaneous Ca(2+) oscillations depend on action potentials mediated by Na(+) channels that are largely TTX-insensitive but blocked by the local anesthetics tetracaine and lidocaine. Collectively, these results corroborate the conclusion that odorant-induced Ca(2+) transients and spontaneous Ca(2+) oscillations in the somata of olfactory receptor neurons closely reflect action potential activity associated with odorant-induced phasic-tonic responses and spontaneous bursting, respectively. Therefore, both types of Ca(2+) signals represent experimentally accessible proxies of spiking.
我们确定了一种新开发的触角切片制备方法,该方法允许在原位条件下使用 Ca(2+) 成像研究棘龙虾嗅觉受体神经元的化学感觉特性。我们表明,化学刺激到达嗅觉受体神经元的树突,但不到达其胞体,并且在胞体中,气味诱导的 Ca(2+) 信号随时间推移足够稳定,可以用大量气味进行刺激。药理学处理用于阐明气味诱导的 Ca(2+) 瞬变和嗅觉受体神经元胞体中自发 Ca(2+) 振荡的来源。这两种 Ca(2+) 信号主要通过电压激活的 Ca(2+) 通道的细胞外 Ca(2+) 内流介导,该通道可被 CoCl2 和 L 型 Ca(2+) 通道阻滞剂维拉帕米阻断。细胞内 Ca(2+) 库对气味诱导的 Ca(2+) 瞬变和自发 Ca(2+) 振荡的贡献很小。气味诱导的 Ca(2+) 瞬变以及自发的 Ca(2+) 振荡依赖于由 Na(+) 通道介导的动作电位,这些动作电位主要对 TTX 不敏感,但被局部麻醉剂甲哌卡因和利多卡因阻断。总的来说,这些结果证实了以下结论:嗅觉受体神经元胞体中的气味诱导 Ca(2+) 瞬变和自发 Ca(2+) 振荡分别与气味诱导的相位紧张反应和自发爆发相关的动作电位活动密切相关。因此,这两种类型的 Ca(2+) 信号都代表了实验可接近的尖峰的替代物。