Extended Data Fig. 3: Retrograde tracing of Purkinje cells targeting ALM.
From: Activity map of a cortico-cerebellar loop underlying motor planning
a. Coronal sections from two example cases showing labeled neurons from retroAAV-Cre injection in ventromedial thalamic nucleus (VM) and Cre-dependent rabies viruses in the fastigial nucleus (Fig. 1f). Dashed line shows boundary of the fastigial nucleus. Purple, TVA positive neurons; blue, rabies positive neurons, that is starter cells. Scale bar, 250 µm. This experiment was repeated in 5 mice with similar results. b. Distribution of all starter cells in the fastigial nucleus. Data from all injection cases (n = 5). Each dot represents one neuron. VM-projecting fastigial neurons are concentrated in the caudal region of the fastigial nucleus. c. Registration of retrogradely labeled Purkinje cells (yellow crosses). Consecutive coronal sections covering the cerebellum are aligned into the Allen Mouse Common Coordinate Framework (CCF). The image shows an example coronal section before and after alignment to the corresponding section of the anatomical template in CCF. Red points indicate control points for b-spline transformation. Annotated Purkinje cells in the coronal section are shown in CCF. d. The number of labeled Purkinje cells is correlated with the number of starter cells in the fastigial nucleus (n = 5 mice). Color codes for individual animals as in e. e. Labeled Purkinje cells in individual injection cases in caudal and dorsal view. Dots show annotated Purkinje cells. The pattern is similar across injection cases. f. Retrograde tracing of Purkinje cells from ventral anterolateral thalamic nucleus (VAL) and dentate nucleus (DN). From left to right, injection schematics; images of injection sites in VAL and DN; images of labeled starter cells in DN, images of labeled Purkinje cells in cerebellar cortex. Histology images from one mouse are displayed. This experiment was repeated in 4 mice with similar results. Scale bars from left to right, 500 µm, 200 µm, 10 µm, 10 µm, 10 µm, 100 µm and 10 µm. g. The number of labeled Purkinje cells is correlated with number of starter cells in DN (n = 4 mice). Color codes for individual animals as in h. h. Labeled Purkinje cells in individual injection cases in caudal and dorsal view. Right, summary of Purkinje cells targeting VAL via DN. Color codes for kernel density estimation of Purkinje cells. i. Comparison of Purkinje cells targeting VM via FN (blue) and Purkinje cells targeting VAL via DN (magenta). j. Comparison of output Purkinje cell density from i in each lobule. Open circles, individual mice (labeling from FN n = 5 mice, DN n = 4 mice). Error bars, s.e.m. across mice.
