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在切片上用电压敏感染料成像观察大鼠脊髓背角内局灶刺激诱发的活动的空间组织。

Spatial organization of activity evoked by focal stimulation within the rat spinal dorsal horn as visualized by voltage-sensitive dye imaging in the slice.

机构信息

Department of Anesthesia and Critical Care, Beth Israel Deaconess Medical Center, Boston, Massachusetts.

Department of Anaesthesia, Harvard Medical School, Boston, Massachusetts.

出版信息

J Neurophysiol. 2019 Oct 1;122(4):1697-1707. doi: 10.1152/jn.00697.2018. Epub 2019 Aug 21.

Abstract

In a prior study using laser scanning photostimulation, we found a pronounced cell type-specific mediolateral asymmetry in the local synaptic connectivity in the superficial laminae of the spinal dorsal horn (Kosugi M, Kato G, Lukashov S, Pendse G, Puskar Z, Kozsurek M, Strassman AM. 591: 1935-1949, 2013). To obtain information on dorsal horn organization that might complement findings from microelectrode studies, voltage-sensitive dye imaging was used in the present study to examine patterns of activity evoked by focal electrical stimulation, in the presence and absence of synaptic blocking agents, at different positions in transverse, parasagittal, and horizontal slices of the dorsal horn of 2- to 3-wk -old male rats. A pronounced difference in responsiveness was found between medial and lateral dorsal horn, in that medial sites in the superficial dorsal horn showed much larger synaptic responses to focal stimulation than lateral sites. This difference appeared to be a result of a difference in the intrinsic elements of the dorsal horn, rather than a difference in the inputs from the white matter, because the stimulus intensities were subthreshold for evoking synaptic responses from stimulation at sites in the white matter, although it is also possible that the greater responsiveness is due, at least in part, to activation of Aβ primary afferent fibers that pass through the medial dorsal horn. The results raise the possibility of differences between medial and dorsal horn that need to be taken into account in the interpretation of studies of dorsal horn organization. We used voltage-sensitive dye imaging to obtain information on spatial aspects of dorsal horn organization that are difficult to examine with single-cell approaches because of the limitations of microelectrode sampling. The most noteworthy finding was a previously unreported, extreme difference between medial and lateral dorsal horn in responsiveness to focal stimulation that appears to result, at least in part, from a greater degree of excitability or local connectivity in medial dorsal horn.

摘要

在之前使用激光扫描光刺激的研究中,我们发现脊髓背角浅层的局部突触连接存在明显的细胞类型特异性的内外侧不对称性(Kosugi M、Kato G、Lukashov S、Pendse G、Puskar Z、Kozsurek M、Strassman AM。591:1935-1949, 2013)。为了获得可能补充微电极研究结果的背角组织信息,本研究使用电压敏感染料成像来检查在 2-3 周龄雄性大鼠的背角横切、矢状旁和水平切片的不同位置,用焦点电刺激诱发的活动模式。在存在和不存在突触阻断剂的情况下,发现内侧和外侧背角之间的反应性存在明显差异,即浅层背角的内侧部位对焦点刺激的突触反应比外侧部位大得多。这种差异似乎是背角固有元素的差异造成的,而不是来自白质输入的差异,因为刺激强度低于在白质部位刺激引起突触反应的阈强度,尽管也有可能更大的反应性至少部分是由于通过内侧背角的 Aβ初级传入纤维的激活。研究结果提出了内侧和背角之间可能存在差异的可能性,在解释背角组织研究时需要考虑到这些差异。我们使用电压敏感染料成像来获取关于背角组织空间方面的信息,这是用单细胞方法难以检查的,因为微电极采样存在局限性。最值得注意的发现是,内侧和外侧背角之间存在以前未报道过的对焦点刺激反应的极端差异,这种差异似乎至少部分是由于内侧背角的兴奋性或局部连接程度更高所致。

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