Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital, McGill University, Montréal, QC H3A 2B4, Canada.
Department of Neurology and Neurosurgery, Montreal Neurological Institute-Hospital, McGill University, Montréal, QC H3A 2B4, Canada
Proc Natl Acad Sci U S A. 2022 Feb 22;119(8). doi: 10.1073/pnas.2107899119.
The development of functional topography in the developing brain follows a progression from initially coarse to more precisely organized maps. To examine the emergence of topographically organized maps in the retinotectal system, we performed longitudinal visual receptive field mapping by calcium imaging in the optic tectum of GCaMP6-expressing transgenic tadpoles. At stage 42, just 1 d after retinal axons arrived in the optic tectum, a clear retinotopic azimuth map was evident. Animals were imaged over the following week at stages 45 and 48, over which time the tectal neuropil nearly doubled in length and exhibited more precise retinotopic organization. By microinjecting GCaMP6s messenger ribonucleic acid (mRNA) into one blastomere of two-cell stage embryos, we acquired bilateral mosaic tadpoles with GCaMP6s expression in postsynaptic tectal neurons on one side of the animal and in retinal ganglion cell axons crossing to the tectum on the opposite side. Longitudinal observation of retinotopic map emergence revealed the presence of orderly representations of azimuth and elevation as early as stage 42, although presynaptic inputs exhibited relatively less topographic organization than the postsynaptic component for the azimuth axis. Retinotopic gradients in the tectum became smoother between stages 42 and 45. Blocking N-methyl-D-aspartate (NMDA) receptor conductance by rearing tadpoles in MK-801 did not prevent the emergence of retinotopic maps, but it produced more discontinuous topographic gradients and altered receptive field characteristics. These results provide evidence that current through NMDA receptors is dispensable for coarse topographic ordering of retinotectal inputs but does contribute to the fine-scale organization of the retinotectal projection.
发育中大脑的功能拓扑结构的发展遵循从最初粗糙到更精确组织图谱的进展。为了研究视顶盖系统中拓扑组织图谱的出现,我们通过在表达 GCaMP6 的转基因 蝌蚪的视顶盖中进行钙成像来进行纵向视觉感受野映射。在 42 期,即在视网膜轴突到达视顶盖后的第 1 天,出现了明显的视方位图。在接下来的一周内,在 45 期和 48 期对动物进行成像,在此期间,视顶盖神经突起的长度几乎增加了一倍,并表现出更精确的视方位组织。通过将 GCaMP6s 信使核糖核酸(mRNA)微注射到双细胞期胚胎的一个卵裂球中,我们获得了双侧镶嵌的蝌蚪,在动物一侧的视顶盖突触后神经元中表达 GCaMP6s,而在对侧穿过视顶盖的视网膜节细胞轴突中表达 GCaMP6s。对视方位图出现的纵向观察表明,早在 42 期就存在方位和仰角的有序表示,尽管与方位轴的突触后成分相比, 突触前输入的拓扑组织相对较少。在 42 期和 45 期之间,视顶盖中的视方位梯度变得更加平滑。通过在 MK-801 中饲养蝌蚪来阻断 N-甲基-D-天冬氨酸(NMDA)受体电导并不能阻止视方位图的出现,但它产生了更不连续的拓扑梯度并改变了感受野特征。这些结果提供了证据,表明 NMDA 受体电流对于视顶盖输入的粗糙拓扑排序是可有可无的,但有助于视顶盖投射的精细组织。