Department of Biology, Program in Neuroscience and Behavior, Wesleyan University, Middletown, Connecticut, United States of America.
PLoS One. 2012;7(2):e31415. doi: 10.1371/journal.pone.0031415. Epub 2012 Feb 21.
We report on a novel mouse in vitro brain slice preparation that contains intact callosal axons connecting anterior cingulate cortices (ACC). Callosal connections are demonstrated by the ability to regularly record epileptiform events between hemispheres (bilateral events). That the correlation of these events depends on the callosum is demonstrated by the bisection of the callosum in vitro. Epileptiform events are evoked with four different methods: (1) bath application of bicuculline (a GABA-A antagonist); (2) bicuculline+MK801 (an NMDA receptor antagonist), (3) a zero magnesium extracellular solution (0Mg); (4) focal application of bicuculline to a single cortical hemisphere. Significant increases in the number of epileptiform events, as well as increases in the ratio of bilateral events to unilateral events, are observed during bath applications of bicuculline, but not during applications of bicuculline+MK-801. Long ictal-like events (defined as events >20 seconds) are only observed in 0Mg. Whole cell patch clamp recordings of single neurons reveal strong feedforward inhibition during focal epileptiform events in the contralateral hemisphere. Within the ACC, we find differences between the rostral areas of ACC vs. caudal ACC in terms of connectivity between hemispheres, with the caudal regions demonstrating shorter interhemispheric latencies. The morphologies of many patch clamped neurons show callosally-spanning axons, again demonstrating intact callosal circuits in this in vitro preparation.
我们报告了一种新型的体外鼠脑切片制备方法,其中包含连接前扣带皮质(ACC)的完整胼胝体轴突。通过在两个半球之间(双侧事件)定期记录癫痫样事件来证明胼胝体连接。通过在体外将胼胝体切开,证明了这些事件的相关性取决于胼胝体。癫痫样事件可以通过以下四种不同方法诱发:(1)灌流应用荷包牡丹碱(GABA-A 拮抗剂);(2)荷包牡丹碱+MK801(NMDA 受体拮抗剂);(3)零镁细胞外液(0Mg);(4)将荷包牡丹碱局部应用于单个皮质半球。在灌流应用荷包牡丹碱时,观察到癫痫样事件的数量显著增加,以及双侧事件与单侧事件的比率增加,但在应用荷包牡丹碱+MK-801 时则没有观察到。只有在 0Mg 中才观察到长癫痫样事件(定义为事件>20 秒)。对单个神经元的全细胞膜片钳记录显示,在对侧半球的局灶性癫痫样事件期间存在强烈的前馈抑制。在 ACC 内,我们发现前 ACC 与后 ACC 之间在半球间连接方面存在差异,后区的半球间潜伏期较短。许多被膜夹夹住的神经元的形态显示出胼胝体跨越的轴突,再次证明了这种体外制备物中完整的胼胝体回路。