The mouse cortico-striatal projectome

Figure 1. Data production and informatics workflow

Data production and informatics workflow. (a) Discrete anterograde tracer injections were made and photomicrographs of the Nissl and tracer-labeled pathways were acquired. (b) Images were imported into in-house software for warping. Fiducial markers were placed on the Nissl image of the sample to be warped, such that they matched the marker pattern of the corresponding atlas Nissl image. The sample was then deformably warped. (c) The warped PHAL channel was segmented and binarized, yielding a monochromatic image of axonal labeling. (d) The segmented image was superimposed onto its corresponding atlas level, in which the CP was divided into grid cells. Superimposed axons in each grid cell were quantified. (e,f) The quantified labeling for each injection was compiled into a spreadsheet and the data were expressed in a matrix with cortical areas along the y axis and cells of the grid along the x-axis (f, top). A community detection algorithm was used to group cortical areas projecting to a common set of cells (f, bottom). (g) The matrix was reordered and color-coded according to community structure. Cells boxed in red depict quantification of segmented image in c. (h) The grid cells were recolored to visualize, in the CP, the spatial arrangement of the communities to which they belong. (i) Communities were subdivided into domains using the centroid, or center of gravity, of each terminal field. The original labeling is shown in red and labeling after 95% cutoff D, which was used for centroid generation, is shown in green. (j) Axonal fields in a community whose centroids were closely spaced defined a domain. (k) Using intensity-weighted centroids as seed points, we parcellated the CP into Voronoi cells, demonstrating the relative domain boundaries.

Figure 2. Community and domain matrices

Visualization of CP communities and domains. Shown are matrices that visualize the CP communities across the CPr (top), CPi (middle) and CPc (bottom). The matrices show cortical injections along the y axis and labeled CP cells along the x axis. For visualization, matrices were reordered and color-coded according to community structure. Corresponding color-coded CP illustrates the spatial arrangement of communities in dorsal striatum. Community color assignments across the CP were coordinated to reflect general projection trends of communities across the structure. For example, projections terminating in the dorsomedial community of the CPi (represented in mustard yellow) generally terminated in the medial community of the CPr (bright yellow) and the dorsal community of the CPc (pastel yellow). Constituents of each community are listed to the left in corresponding color-coded boxes. Cortical area abbreviations: ACAd, anterior cingulate area, dorsal; ACAv, anterior cingulate area, ventral; AId, agranular insular area, dorsal; AIp, agranular insular area, posterior; AIv, agranular insular area, ventral; AUDd, auditory area, dorsal; AUDp, primary auditory area; AUDv, primary auditory area, ventral; ECT, ectorhinal area; ENTl, entorhinal area, lateral; ENTm, entorhinal area, medial; GU, gustatory area; ILA, infralimbic area; MOp, primary motor area; MOp tr, MOp trunk; MOp ll, MOp lower limb; MOp ul, MOp upper limb; MOp m/i, MOp inner mouth; MOp m/o, MOp outer mouth; MOs, secondary motor area; MOs tr, MOs trunk; MOs ll, MOs lower limb; MOs ul, MOs upper limb; MOs m/i, MOs inner mouth; MOs m/o, MOs outer mouth; MOs-fef, MOs frontal eye field; ORBl, orbitofrontal area, lateral; ORBm, orbitofrontal area, medial; ORBvl, orbitofrontal area, ventrolateral; PERI, perirhinal area; PIR, piriform cortex; PL, prelimbic area; PL dorsal (L6), layer 6 of dorsal PL; PTLp, posterior parietal association area; Rostral MOs (P1), rostral MOs pole 1; Rostral MOs (P2), rostral MOs pole 2; RSPagl, retrosplenial area, agranular; RSPd, retrosplenial area, dorsal; RSPv, retrosplenial area, ventral; SUBd, subiculum, dorsal; SSp, primary somatosensory area; SSp-tr, SSp trunk; SSp-ll, SSp lower limb; SSp-ul, SSp upper limb; SSp-m/i, SSp inner mouth; SSp-m/o, SSp outer mouth; SSp-bfd, SSp barrel field; SSp-bfd tr, SSp-bfd trunk; SSp-bfd ll, SSp-bfd lower limb; SSp-bfd ul, SSp-bfd upper limb; SSp-bfd m/o, SSp-bfd outer mouth; TEa, temporal association area; VISal, visual area, anterolateral; VISam, visual area, anteromedial; VISl, visual area, lateral; VISp, primary visual area; VISpl, visual area, posterior lateral; VISpm, visual area, posterior medial; VISC, visceral area.

Figure 3. Summary of community and domain nomenclature and list of abbreviations

Summary of community and domain nomenclature and CP domain parcellations, (a) Summary of communities and domains for the rostral, intermediate and caudal CP are presented in a hierarchical tree. The nomenclature follows the CPx.y.z convention, where x denotes the level (rostral, intermediate or caudal), y the community and z the domain. (b) Overview of connectivity-based CP domains defined with Voronoi parcellation. A total of 29 functional domains were identified in the rostral (CPr, left), intermediate (CPi, middle) and caudal (CPc, right) levels of the CP. Three domains that were identified based primarily from manual inspection of the data are not shown: the CPc.vl and CPc.v.vm, which are part of the CPc.v and the CPc.ext, located at the tail end of the CP.

Figure 4. Somatotopic map of cortico-striatal projections to the CP

Somatotopic map of cortico-striatal projections to the CPi. (a) Raw data showing somatic sensorimotor areas topographically projecting to different domains in the CPi.dl and CPi.vl communities. Each sensorimotor cortical area representing a unique body region sent converging input to their corresponding CP domain. (b) Dorsal view of the somatic sensorimotor cortical map color-coded according to their domains (modified from ref. 1) and maximum projection of their CP termination fields (bottom) showing distinct, partially overlapping domains. (c) Map depicting connections from body specific regions in SSp (row 1), MOp (row 2), SSp-bfd (row 3) and MOs (row 4) to CP, illustrating dorsal-ventral representation of trunk, lower limb, upper limb, inner mouth and outer mouth observed in a. The map also demonstrates general trend of projections across the rostrocaudal CP. (d) Centroids of cortical areas grouped into CPi.dl and CPi.vl communities. Centroids of body-specific SSp, SSp-bfd, MOp and MOs topographically arranged and subdivided communities into domains. Notably, centroids aligned with the raw data in a and reconstructions in c. PTLp rostral was grouped into the CPi.dl community. However, according to its centroid (denoted with by the asterisk) and raw data (Supplementary Fig. 6d), it more appropriately grouped with structures in a dorsomedial domain. Cortical areas projecting to each domain are listed in boxes color-coordinated with centroids (box stroke color denotes community). (e) Illustration of five parallel, relatively segregated somatic sensorimotor striatal networks. All SS and MO nodes in each subnetwork dedicated to a specific body region are reciprocally connected (Supplementary Fig. 5e and ref. 1) and sent converging projections to CPi domain representing their corresponding body region. (f) Interdigitation of projection terminals in CPi trunk domain (CPi.dl.d) revealed by double anterograde injections MOs/MOp trunk regions. (g) Projections from rostral MOs (pole 2) arborized in the central CP, avoiding the periphery. (h) FG, CTb 647 and 549 injected in trunk, lower limb and upper limb/mouth CP domains retrogradely labeled layer V CP-projecting cortical neurons in their corresponding SSp and MOp body subregions, as well as SSs and VISC.

Figure 5. Cortical projections to the dorsomedial and ventromedial CPi communities

Cortical projections to the dorsomedial and ventromedial CPi communities. (a) Dorsal and lateral views of cortical regions in the medial and lateral cortical subnetworks, respectively (modified from ref. 1). Areas in the medial cortical subnetwork projected to domains in CPi.dm, whereas those in the lateral cortical subnetwork projected to CPi.vm domains. Colors of cortical areas denote domain assignments. Overlap of reconstructed pathways of areas projecting CPi.dm and CPi.vm highlights unique domains. (b) Triple anterograde tracer injections in VISam (AAV-RFP), ORBvl (PHAL) and PL (AAV-GFP) revealed the relative boundaries of CPi.dm.d, CPi.dm.im and CPi.dm.cd. (c) Double injections of BDA and PHAL in VISl and TEa showed segregation of CPi.dm.dm and CPi.dm.im. (d) Raw data from representative cases of cortical areas projecting to CPi.vm illustrate the CPi.vm.vm and CPi.vm.v domains. (e) Double injections of AAV-GFP (PTLp caudal medial) and PHAL (ACAd) highlight interdigitating boundary between CPi.dm.d and CPi.dm.dl. (f) Centroids of cortical areas projecting to CPi.dm and CPi.vm displayed high topography and identified domains. Cortical areas projecting to each domain are listed in a box that is color-coordinated with the centroids (stroke color of box denotes community). The algorithm grouped LA with cortical structures projecting to the CPi.dm; however, its centroid (denoted with an asterisk) placed it outside of the five domains of this community and into CPi.vm.vm. (g) Wiring schema depicts connections among cortical areas in the medial cortical subnetwork dedicated to transferring visual, auditory and some somatosensory (trunk/lower limb) information to the ORBvl through interconnected nodes involving the VIS, ACA, RSP, PTLp and AUD1. Nodes in this subnetwork project to domains in the CPi.dm, forming a medial cortical-dorsomedial striatal network. Diagram also depicts the anterolateral cortical-ventromedial striatal subnetwork. The anterolateral cortical subnetwork (that is, AI, GU, VISC, PIR) was heavily connected with PL and ILA. Downstream projections of each of these nodes generally target the domains of the CPi.vm.

Figure 6. Cortico-striatal projections in the CPr and CPc

Convergence and reconfiguration of cortico-striatal projections in CPr and CPc. (a) Map showing reconstructions of projections populated according to CPr domains (that is, row 1 shows projections from all cortical areas that send input to CPr.l.ls; row 2 depicts projections from cortical areas to CPr.l.vm). Reconstructions demonstrate CPr domains (column 1) and show general trend of projections through CPi and CPc. For example, projections to lateral CPi tended to terminate in lateral CPr, whereas projections terminating in the medial CPi terminated in the medial CPr. (b) Map showing reconstructions of projections according to CPc communities and delineating the dorsal, intermediate and ventral regions of CPc (column 3). The termination pattern of cortical areas clustered into these caudal communities across CPi and CPr are also depicted (that is, projections terminating in dorsal CPi terminate in dorsal CPc, whereas those terminating in lateral CPi terminated in intermediate parts of CPc. (c) Raw data from representative cases showing projections to CPr.l.ls and CPr.l.vm. (d) FG and CTb 647 injections into the lateral strip (CPr.l.ls) and medial/intermediate ventral CP (CPr.m/CPr.imv), respectively, validated cortical projections to those domains and communities. FG retrogradely labeled neurons in PL, ORBvl, LA and mBLAa, whereas CTb labeled neurons in MOs, SSp, SSs. Right, magnification of retrograde labeling. (e) Representative cases demonstrating raw labeling to domains in CPc.d. (f) GU injection revealed the ventrolateral domain of CPc.v (CPc.v.vl). Right, magnification of labeling in boxed region. CEA, central amygdalar nucleus; GPe, globus pallidus external part; BLA, basolateral amygdalar nucleus, anterior part. (g) Injections in VISC caudal produce unique labeling in CPc.v.vm (magnified on right). An injection in the thalamus validated this CPc.v.vm domain. Thalamic injection validateD the general projection trends observed across the rostral-caudal extent of the CP for ventral domains (that is, tendency of projections to CPi.vm.v to terminate in ventral domains of CPr and CPc). Right, magnification of labeling in inset magnified at right.

Figure 7. Reconfiguration and network model of the CP

Network reconfiguration across the CP. (a) A hypothetical functional model of the CP domains based on all cortical afference to specific domains identified across the CP. (b) Double anterograde injections of AAV-GFP in SSp-bfd mouth/SSs and PHAL in MOp ul showed segregation of somatic sensorimotor projection terminals in their corresponding domains in the CPi and CPc and intermixing of their axons in CPr, demonstrating reconfiguration of these subnetworks across the CP. (c) Schematic representation of subnetwork reorganization across the CP. Although relatively segregated in CPi, all sensorimotor subnetworks representing unique body information sent converging inputs to a single domain in the CPr. At the CPc, inputs from the somatic sensorimotor nodes terminated in the same community as inputs from the medial (dorsal CPc) and lateral (ventral CPc) subnetworks, providing greater opportunity for interaction across different subnetworks. (d) AUDp projections produced a unique dense terminal field in caudal parts of the CP, where projections from most cortical areas were absent (top left). AUD projections to this domain, termed the caudal extreme (CPc.ext), were validated with a CTb injection, which retrogradely labeled neurons in AUDv (top right). Bottom, magnified images of boxed region showing dense topographic projections from AUDp and TEa in the CPc.ext. CEA, central amygdalar nucleus; BLAa, basolateral amygdalar nucleus, anterior part. (e) FG and CTb 647 tracer injections in the dorsal and ventral CPc substantiate the cortical projections from ACA, MOs-fef, RSP, VIS and ECT to CPc.d and from SSp-m to CPc.v. Right, magnified images of labeling. (f) Projections from ENTl to CPr, CPi, and CPc. Note the broad projections spanning across all CPi.dm and CPi.vm domains in CPi.

Figure 8. Domain-specific pathological connections in zQ175 and MAO AB KO mice

Domain-specific pathological connections in zQ175 and MAO A/B KO mice. (a) Boxplot showing domain-specific loss of signal intensity from the MOs upper limb in zQ175 mice compared with WT littermates in ipsilateral CP (zQ175, n = 13; WT, n = 15). Boxes depict the range between the upper and lower quartiles, whiskers depict the maximum and minimum values, and the bar indicates the median value. Red denotes domains in which connections were affected. (b,e) Pixelgrams illustrating differences in signal intensity detected between zQ175 and MAO A/B KO mice and their WT littermates in each 35 x 35 pixel cell. Red denotes a reduction in signal compared with WT, whereas blue indicates an increase. Saturation of color scales with percent change. (c) Visualization of the bar graphs showing domain-specific signal intensity reductions from MOs upper limb and ORBvl in zQ175 (left) and MAO A/B KO (right) mice, respectively. (d) Boxplots showing domain-specific loss of signal intensity from the ORBvl in MAO A/B KO mice compared with WT in ipsilateral (left) and contralateral (right) CP (MAO A/B, n = 10; WT, n = 6). Only domains in which significant changes were detected are shown. (f) Boxplots of differences in the span of MOs upper limb and ORBvl projection fields in the CP in zQ175 and MAO A/B KO mice compared with WT. (g) Boxplots showing the reduction in signal intensity in the entire ipsilateral CP from the MOs upper limb (left) and ORBvl (right) in zQ175 and MAO A/B mice, respectively. Asterisk denotes significant difference (P < 0.05) detected by two-tailed t test with Welch’s correction. Intensity is measured as total number of labeled pixels. Span is measured as total number of labeled cells

Supplementary figure 1. Topography and termination patterns of cortico-striatal projections

(a) Manual inspection of ~150 tracer-labeled cortico-striatal pathways revealed highly topographic projection patterns across the rostral-caudal extent of the CP. Tracer injections (column 1) were relatively small and confined within anatomically delineated cortical regions thereby producing discrete and specific labeling across the CP (columns 2-7). Injections in different cortical areas also produced a variety of termination patterns within the projection fields. Magnified images of the labeling at the intermediate CP is presented in the final column and demonstrates the different axonal terminal patterns. Some show dense terminal fields surrounded by diffuse labeling, others are arranged as horizontal or vertical stripes, and some are confined to the center parts of the CP while most others run along the periphery. (b) The large majority of the anterograde tracing data was confirmed with retrograde tracer injections placed in the CP. Injections of four different tracers infused into different regions within the CP retrogradely labeled different populations of cortical cells validating unique cortical projection sources to different parts of the CP. They also revealed the laminar organization of the CP projecting neurons.

Supplementary figure 2. Triple anterograde tracing and tracer-dependent labeling

Supplementary figure 3. Intensity, span and labeling density index values

(a) Graphical representation of intensity and span as complementary measures to quantify density of CP labeling from individual cortical sources. Intensity is the sum of the labeled pixels across the CP and thereby indexes the concentration of labeling. Span is the count of cells containing labeling and therefore measures how diffuse the labeling is throughout the CP. Projections from the ACAd label a large majority of the 35 x 35 pixels within each cell of the CP (blue; intensity=4,046,768) and therefore show a higher intensity value than projections from the SS rostral, whose projections show far less labeling of pixels (red; intensity=161,560). On the other hand, the ACAd injection labels far fewer cells compared to projections from the SSs rostral and therefore has a much lower span value (1,453 compared to 2,406 for SSs rostral). The Labeling Density Index (LDI) summarizes these labeling properties in a single value, which is the ratio of the intensity to span. LDI values greater than 1.0 are designated as concentrated, with lesser LDI values defined as diffuse. (b-d) Intensity, span and LDI values for each cortical area are presented for the rostral, intermediate and caudal CP.

Supplementary figure 4. Table of communities and domains and their cortical constituents

(a) Table showing all cortical areas that project to unique communities and domains within the intermediate CP. Cortical areas are color coded according to their projections to domains and are the same as their centroid colors (i.e., Fig. 5f). (b-c) Table of cortical areas that project to unique communities and domains within the rostral and caudal CP. Colors for cortical areas are the same as those used for the intermediate CP to highlight the higher degree of projection integration from different cortical areas across the CP

Supplementary figure 5. Somatic sensorimotor cortical injections, projections and intracortical connections

(a) Injection locations for all somatic sensorimotor cortical areas representing unique body subregions. Generally within each somatic sensorimotor cortical area, there was a caudomedial to rostrolateral representation of trunk, lower limb, upper limb, inner mouth and outer mouth. The literature available for somatotopic cortical maps largely support these findings, although this is the first report to identify all body subregions for all somatic sensorimotor cortical areas61 (also see Woolsey et al., Res. Publ. Assoc. Res. Nerv. Ment. Dis.,1952; Hall & Lindholm, Exp. Brain Res., 1974; Welker, J. Comp. Neurol., 1976; Muakkassa & Strick, Brain Res., 1979; Donoghue & Wise, J. Comp. Neurol., 1982; Tanji & Kurata, J. Neurophysiol., 1982; Godschalk et al., Exp. Brain Res.,1984; Neafsey et al., Brain Res., 1986; Mitz & Wise, J. Neurosci., 1987). (b) Panels show retention of the CP somatotopy in more caudal parts of the CP (i.e., -0.28 mm from bregma). (c) The specificity of injections within the SSp-body subregions was validated by examining their thalamic and brainstem projections. Leftmost panels are schematic representations of the somatosensory thalamus (top) and brainstem (bottom) modified from57 (also see Nord, J. Comp. Neurol., 1967). The thalamus schematic shows the approximate location of body representations within the ventral posteromedial (VPM) and ventral posterolateral (VPL) thalamic nuclei. Injections into the SSp-tr, ll, ul and m regions showed clear topographic projections to their corresponding VPM and VPL regions. Trunk representations were in dorsal regions, followed by lower limb which were lateral to upper limb representations, with orofacial representations in the most ventral and medial parvocellular regions57-60 (also see Emmers, J. Comp. Neurol., 1965). The somatosensory brainstem schematic shows the approximate location of the body representations in the spinal trigeminal, the cuneate and gracile nuclei. Consistent with these brainstem body representations, SSp-ll labeled the gracile nucleus, the SSp-ul the cuneate, and the SSp-m the dorsal parts of the spinal trigeminal (SPV) nucleus with SSp-m/o represented dorsal and lateral to the SSp-m/i regions61 (also see Nord, J. Comp. Neurol., 1967; Kruger et al., J. Neurophysiol.,1961; Kruger & Michel, Exp. Neurol., 1962). The cortical location of the SSp-m/i and SSp-m/o also agree with data showing that SSp representations of the tongue are generally more rostral and lateral than those of the lips in rats62 and monkeys63. (d) Specificity of SSp-bfd injections was also validated by observing their topographic projections to the medial part of the posterior thalamus (POm), to the VPM and to approximately the ventral third of the spinal trigeminal, precisely where whisker representations reside57, 58,64 (also see Nord, J. Comp. Neurol., 1967). (e) Panels showing strong intra-cortical connectivity among all somatic sensorimotor cortical regions for upper limb. Figure adapted from 1. (f, left panels) Mouth somatic sensorimotor region projections to the CPi.vl.v and vt were validated with a FG injection placed within these domains. FG retrogradely labeled cells within the MOp and SSp regions for mouth, but did not label cells within SSp and SSp-bfd regions for trunk. (f, right panels) CTb 647 injection in CP trunk region substantiated its inputs from somatic sensorimotor trunk regions. Cells were labeled in the trunk regions within the MOp, SSp and SSp-bfd.

Supplementary figure 6. Cortical projections to dorsomedial (CPi.dm) and ventromedial (CPi.vm) CPi

(a) A thalamic injection shows labeling of the CPi.vl.cvl substantiating the domain, but also validating the projection trend for this domain across the CP. Both rostral MOs (pole 2) and thalamic projections to the CPi.vl.cvl terminate in the lateral strip domain of the CPr and in the intermediate part of the CPc. (b) Centroids for the cortical areas that project to the central CP domains (denoted by *). Cortical areas that project to each center domain are listed next to the centroids. (c) Raw data showing cortical projections from the ORBvl and ORBl to the CPi center domain CPi.dm.cd and from the rostral MOs (pole 1) to the CPi.vm.cvm. (d) Raw data showcasing representative projections to the dorsomedial CPi domains. (e) Connectivity of the MOs-fef (ACAd/MOs region) compared to those of the VISam and from the somatomotor region for lower limb. Unlike the MOs ll, the MOs-fef is connected with other visual cortical areas like the VISam, VISp and RSPv. Similar to the VISam, the MOs-fef is also connected with visual thalamic nuclei like the lateral dorsal (LD) and projects to the CPi.dm.d, the domain of the CP that receives visual and spatial information from the VISam, VISal, ACAv and PTLp. The MOs ll on the other hand projects to the somatosensory lateral part of the CPi. (f) Topographically arranged projections from the VISp and VISpm to the CPi.dm.dm domain. The terminal field from the VISpm is typically in between the two patches of segregated terminals from the VISp. (g) Double anterograde tracers display the boundary between the CPi.dm.im and CPi.vm.vm domains. (h-j) Injections in different thalamic nuclei validate the CPi.dm.d, CPi.dm.dm and CPi.vl.v domains of the intermediate CP. (k) A CTb 647 injection in the CPi.dm.dl and CPi.dl.d (trunk region) retrogradely labels cells in the ACAd, MOp tr, SSp-tr and PTLp caudal lateral confirming their projections to these domains.

Supplementary figure 7. Cortical projections to the rostral (CPr) and caudal (CPc) CP

(a) Color-coded CP in the left panel shows the four CPr communities, which are also faithfully represented by the centroids (right panels). Centroid color assignment from the intermediate CP was maintained in the rostral and caudal CP to highlight the higher degree of cortical convergence that occurs at these CP divisions. This precluded identification of smaller domains within the CPr communities with the exception of the CPr.l, which was subdivided into two domains: the lateral strip (CPr.l.ls) and ventromedial (CPr.l.vm). All cortical areas projecting to each community are listed in a box next to the centroids. Boxes are color coded according to community structure. (b) Raw data demonstrating the domains of the CPr. (c) Thalamic injections validating the CPr.m and CPr.imd. (d) Raw images of representative cases for the dorsal (left) and intermediate (right) communities in the CPc. (e) Centroids representing dorsal (top; CPc.d), and intermediate (CPc.i) and ventral (CPc.v) (bottom) communities. The SSp-ll was grouped with the CPc.d communities by the algorithm, but its centroid (green, denoted with an *) fell in between the dorsal and intermediate communities. Based on its raw labeling and the centroid, the SSp-ll was grouped with cortical areas projecting to the CPc.i. All cortical areas projecting to each CPc domain are listed next to the centroids. Boxes are colored according to community assignment.

Supplementary figure 8. MOs ul, MOp ul and ORBvl injections in zQ175 and MAO A/B KO mice

(a) Consistent placement of injections in layer V of MOs upper limb region in zQ175 and WT subjects. Boxed regions are magnified and show individual neurons that absorbed the tracer at the center of the injection site. (b) Regression scatterplots showing a positive correlation between injection site size and label intensity in the CP for MOs upper limb, MOp upper limb and ORBvl injections as measured by Pearson’s r. Injections in MOp upper limb showed a trend toward a significant correlation. [MOs ul: r2=0.1724, P=0.028; ORBvl: r2=0.3788, P=0.012; MOp ul: r2=0.1018, P=0.312]. (c) ORBvl injection captured with higher exposure setting to reveal labeling shows overexposed PHAL injection site. The same injection captured with lower exposure times reveals the number of neurons that absorbed the tracer for transport. (d) Boxplots showing that no group differences were detected in injection site size between zQ175 and MAO A/B KO mice and their respective WT littermate controls. Boxes depict the range between the upper and lower quartiles, whiskers depict the maximum and minimum values, and the bar indicates the median value. [Injection site, zQ175 or MAO Mean/SEM, WT Mean/SEM, P value, t value, degrees of freedom; MOs ul: 26.69/2.986, 21.60/2.640, 0.214, 1.278, 24; MOp ul: 36.00/7.452, 42.00/7.545, 0.585, 0.5658, 9; ORBvl: 25.10/3.024, 26.83/5.043, 0.776, 0.2948, 8]. * denotes significant difference (P<0.05) detected by two-tailed t test with Welch’s correction.

Supplementary figure 9. Cortico-striatal projections in zQ175 and MAO A/B KO mice

(a-b) Raw CP labeling resulting from injections placed in the MOs upper limb of zQ175 mice and in the ORBvl of MAO A/B KO mice, respectively. (c) Boxplot of projections from MOs upper limb to contralateral CP in zQ175 animals and their WT littermates. Boxes depict the range between the upper and lower quartiles, whiskers depict the maximum and minimum values, and the bar indicates the median value. [Domain, Mean/SEM zQ175 group, Mean/SEM WT group, P value, t value, degrees of freedom: i.vl.imv (ul), 9437/1417, 16960/2595, 0.019, 2.543, 21; i.dl.imd (ll), 5284/927, 10070/1609, 0.017, 2.578, 22; i.dl.d (tr), 1801/376, 3104/538.6, 0.059, 1.984, 24; i.dm.d, 59.16/28.27, 125.6/97.46, 0.522, 0.6543, 16; i.dm.dl, 316.1/123.4, 414.9/156.4, 0.6243, 0.4959, 25; i.dm.im, 25.83/8.460, 127.6/104.4, 0.347, 0.972, 14; i.dm.cd, 466.3/153.7, 631.3/182.8, 0.496, 0.6907, 25; i.dm.dm, 29.55/21.95, 56.25/51.39, 0.639, 0.4779, 18; i.vm.vm, 308.0/131.9, 607.6/178.5, 0.190, 1.350, 24; i.vm.v, 795.6/278.1, 2047/725.1, 0.126, 1.611, 17; i.vm.cvm, 811.7/233.2, 1609/345.2, 0.068, 1.914, 23; i.vl.v (m/i), 7706/1189, 12110/1844, 0.057, 2.007, 23; i.vl.vt (m/o), 5230/663, 10890/2551, 0.048, 2.149, 15; i.vl.cvl, 3625/566, 7937/1359, 0.009, 2.930, 18]. (d) Boxplot of projections from MOp upper limb to ipsilateral CP in zQ175 and WT animals (zQ175, n=6; WT, n=6). No significant differences in normalized signal intensity values were detected in ipsilateral or contralateral (data not shown) domains. [Ipsilateral: i.vl.imv (ul), 6384/1663, 4446/1353, 0.390, 0.9039, 9; i.dl.imd (ll), 4111/1119, 2779/853.9, 0.369, 0.9464, 9; i.dl.d (tr), 1453/629.2, 852.4/216.3, 0.401, 0.9030, 6; i.dm.d, 0.4667/0.3670, 17.85/15.35, 0.309, 1.132, 5; i.dm.dl, 449.9/304.9, 229.8/139.1, 0.536, 0.6567, 6; i.dm.im, 1.424/1.352, 1.235/0.7757, 0.907, 0.1208, 7; i.dm.cd, 80.18/31.34, 51.14/29.84, 0.519, 0.6709, 9; i.dm.dm, 0.3299/0.3299, 0.1421/0.1159, 0.2515, 1.236, 8; i.vm.v, 863.6,/332.0, 544.3/149.2, 0.414, 0.8773, 6; i.vm.cvm, 646.7/228.7, 248.5/105.2, 0.158, 1.582, 7; i.vl.v (m/i), 3807/1336, 3188/785.4, 0.700, 0.3991, 8; i.vl.vt (m/o), 3484/1156, 2894/680.2, 0.671, 0.4403, 8; i.vl.cvl, 3703/1246, 1653/284.8, 0.170, 1.604, 5; Contralateral: i.vl.imv (ul), 4606/1480, 3967/1179, 0.743, 0.3378, 9; i.dl.imd (ll), 3435/1328, 2743/591.0, 0.651, 0.4758, 6; i.dl.d (tr), 1116/535.8, 725.9/153.7, 0.516, 0.6995, 5; i.dm.d, 3.693/2.297, 7.712/3.790, 0.391, 0.9068, 8; i.dm.dl, 128.7/87.71, 114.4/35.54, 0.885, 0.1506, 6; i.dm.im, 1.270/0.7481, 4.077/2.045, 0.245, 1.289, 6; i.dm.cd, 78.44/42.89, 62.95/12.95, 0.743, 0.3459, 5; i.dm.dm, 0.3125/0.2096, 1.806/1.596, 0.396, 0.9279, 5; i.vm.vm, 422.0/200.7, 301.3/73.54, 0.593, 0.5649, 6; i.vm.v, 437.1/143.1, 561.0/122.0, 0.526, 0.6590, 9; i.vm.cvm, 85.75/53.76, 60.22/12.83, 0.664, 0.4619, 5; i.vl.v (m/i), 4533/1687, 4059/1589, 0.843, 0.2042, 9; i.vl.vt (m/o), 3725/1384, 3715/1391, 0.996, 0.004827, 9; i.vl.cvl, 1715/712.8, 1490/138.3, 0.769, 0.3101, 5]. (e) Comparisons of MOp upper limb projection field span in ipsilateral and contralateral CP between zQ175 and WT mice. [Ipsilateral: 4432/407, 4208/241, 0.648, 0.4741, 8; Contralateral: 1017/31, 1044/39, 0.606, 0.5347, 9]. (f) MOs upper limb and ORBvl projection field span in contralateral CP of zQ175 (left) and MAO A/B KO (right) mice compared to their respective WT littermates. [MOs ul: 4843/302, 5187/296, 0.423, 0.8142, 25; ORBvl: 1018.182, 2314/370, 0.016, 3.142, 7]. * denotes significant difference (P<0.05) detected by two-tailed t test with Welch’s correction. Intensity is measured as the total number of labeled pixels. Span is measured as the total number of labeled cells.