Delaney K, Davison I, Denk W
Department of Biological Sciences, Simon Fraser University, 8888 University Drive, Burnaby, B.C. V5A 1S6, Canada.
Eur J Neurosci. 2001 May;13(9):1658-72. doi: 10.1046/j.1460-9568.2001.01545.x.
We measured Ca2+ concentration, [Ca2+], transients in mitral cell distal apical dendritic tufts produced by physiological odour stimulation of the olfactory epithelium and electrical stimulation of the olfactory nerve (ON) using two-photon scanning and conventional wide-field microscopy of Ca2+-Green-1 dextran in an in vitro frog nose-brain preparation. Weak or strong ON shock-evoked fluorescence transients always had short latency with an onset 0-10 ms after the onset of the bulb local field potential, rapidly increasing to a peak of up to 25% fractional fluorescence change (DeltaF/F) in 10-30 ms, were blocked by 10 microM CNQX, decaying with a time constant of about 1 s. With stronger ON shocks that activated many receptor axons, an additional, delayed, sustained AP5-sensitive component (peak at approximately 0.5 s, up to 40% DeltaF/F maximum) could usually be produced. Odour-evoked [Ca2+] transients sometimes displayed a rapid onset phase that peaked within 50 ms but always had a sustained phase that peaked 0.5-1.5 s after onset, regardless of the strength of the odour or the amplitude of the response. These were considerably larger (up to 150% DeltaF/F) than those evoked by ON shock. Odour-evoked [Ca2+] transients were also distinguished from ON shock-evoked transients by tufts in different glomeruli responding with different delays (time to onset differed by up to 1.5 s between different tufts for the same odour). Odour-evoked [Ca2+] transients were increased by AMPA-kainate receptor blockade, but substantially blocked by AP5. Electrical stimulation of the lateral olfactory tract (5-6 stimuli at 10 Hz) that evoked granule cell feedback inhibition, blocked 60-100% of the odour-evoked [Ca2+] transient in tufts when delivered within about 0.5 s of the odour. LOT-mediated inhibition was blocked by 10 microM bicuculline.
我们在体外青蛙鼻脑制备物中,使用双光子扫描和Ca2+-Green-1葡聚糖的传统宽视野显微镜,测量了嗅上皮的生理性气味刺激和嗅神经(ON)电刺激在二尖瓣细胞远端顶端树突丛中产生的Ca2+浓度[Ca2+]瞬变。弱或强的ON电击诱发的荧光瞬变潜伏期总是很短,在球状体局部场电位开始后0 - 10毫秒开始,在10 - 30毫秒内迅速增加到高达25%的分数荧光变化(ΔF/F)峰值,被10微摩尔的CNQX阻断,以约1秒的时间常数衰减。对于激活许多受体轴突的更强ON电击,通常会产生一个额外的、延迟的、持续的AP5敏感成分(约0.5秒达到峰值,最大ΔF/F高达40%)。气味诱发的[Ca2+]瞬变有时显示出在50毫秒内达到峰值的快速起始阶段,但总是有一个持续阶段,在起始后0.5 - 1.5秒达到峰值,无论气味的强度或反应的幅度如何。这些瞬变比ON电击诱发的瞬变大得多(高达150%ΔF/F)。气味诱发的[Ca2+]瞬变还与ON电击诱发的瞬变不同,不同肾小球中的树突丛以不同的延迟做出反应(对于相同气味,不同树突丛之间的起始时间差异高达1.5秒)。气味诱发的[Ca2+]瞬变在AMPA-海人藻酸受体阻断时增加,但基本上被AP5阻断。外侧嗅束的电刺激(10赫兹下5 - 6次刺激)诱发颗粒细胞反馈抑制,当在气味出现后约0.5秒内施加时,可阻断60 - 100%的气味诱发的树突丛中的[Ca2+]瞬变。LOT介导的抑制被10微摩尔的荷包牡丹碱阻断。