在尝试利用海马组织中的局部场电位测量来开发用于超声神经调节的体外模型过程中所面临的挑战及汲取的经验教训。
Challenges and lessons learned in attempts to develop in vitro models for ultrasound neuromodulation using local field potential measurements in hippocampal tissue.
作者信息
Prieto Martin Loynaz, Madison Daniel V, Maduke Merritt
机构信息
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California.
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, California.
出版信息
Biophys J. 2025 Jun 30. doi: 10.1016/j.bpj.2025.06.039.
Ultrasound neuromodulation is a rapidly developing tool for noninvasive control of brain activity. An in vitro model recapitulating the effects of ultrasound on neural tissue in vivo would be extremely valuable in guiding the development of this tool for optimal implementation. Yet, there are relatively few studies of ultrasound on neural activity in vitro. Here, we describe our attempts to measure neuromodulatory outcomes using local field potential measurements in two in vitro models of hippocampal activity. First, we measured the effects of ultrasound at 1 MHz and 100-600 kPa on a mouse hippocampal in vitro model of sharp wave ripples. Our primary protocol involved brief ultrasound pulses delivered at intervals shorter than the mean interval between sharp wave ripple events, with other stimulus protocols tested with small sample size. No set of parameters produced detectable effects on the amplitude or frequency of sharp wave ripples. We considered whether missing synaptic connections or the relatively small volume in brain slices might account for the lack of effect in our experimental setup. To test these hypotheses, and to examine ultrasound's effects in another system, we measured the effects of ultrasound on theta oscillations in the intact rat hippocampus in vitro. We found that ultrasound at 1 MHz and 500 kPa, applied continuously for 2 s, produced no detectable effects on the amplitude or frequency of in vitro theta oscillations. Finally, we considered a novel mechanism for ultrasound's effects on neural activity, in which acoustic pressure causes microscale phase transitions in the pores of ion channels, such as nicotinic receptor channels, that exhibit hydrophobic gating. To test this hypothesis, we repeated our experiments on the intact hippocampus in the presence of 5 μM nicotine; however, as with the other experimental systems, we found no detectable effects of ultrasound in our experimental setup.
超声神经调节是一种用于无创控制大脑活动的快速发展的工具。构建一个能概括超声在体内对神经组织作用效果的体外模型,对于指导该工具的优化开发极具价值。然而,关于超声对体外神经活动的研究相对较少。在此,我们描述了在两种海马体活动的体外模型中,尝试通过局部场电位测量来测定神经调节结果的过程。首先,我们在1兆赫和100 - 600千帕的条件下,测量了超声对小鼠海马体体外尖波涟漪模型的影响。我们的主要实验方案包括以短于尖波涟漪事件平均间隔的时间间隔施加短暂的超声脉冲,同时也用小样本量测试了其他刺激方案。没有一组参数能对尖波涟漪的幅度或频率产生可检测到的影响。我们考虑了缺失的突触连接或脑片相对较小的体积是否可能是导致我们实验装置中未出现效果的原因。为了验证这些假设,并在另一个系统中研究超声的作用,我们测量了超声对完整大鼠体外海马体中θ振荡的影响。我们发现,在500千帕下连续施加2秒的1兆赫超声,对体外θ振荡的幅度或频率没有产生可检测到的影响。最后,我们考虑了一种超声对神经活动作用的新机制,即声压会在离子通道(如烟碱受体通道)的孔隙中引起微观尺度的相变,这些离子通道表现出疏水门控特性。为了验证这一假设,我们在存在5微摩尔尼古丁的情况下,对完整海马体重复了我们的实验;然而,与其他实验系统一样,我们在实验装置中未发现超声有可检测到的影响。
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