Venus expression in the nervous system other than the retina in Aldoc-Venus mice.

<p>A, Ampullas of the vertical semicircular canals. Whole mount preparation. B and C, Dorsal cochlear nucleus in a parasagittal section. High magnification (C) shows a labeled cartwheel cell. D–E, Spinal cord. F–K, coronal sections of the brain at different levels from the hindbrain to olfactory bulb. Squares (in B, E, H–K) indicate the areas that are magnified in separate panels. L–P, Double labeling with DAPI in dorsal root ganglion, CA1 of the hippocampus, olfactory bulb, corpus callosum and cerebral cortex, respectively, in medium magnification. Q, Confocal photomicrograph of double labeling of Venus and DAPI in an Aldoc-positive area of the cerebellar cortex. All PCs expressed Venus strongly at their soma, dendrites and axon terminals. Paraflocculus in a coronal section. Filled arrowheads indicate somata of Bergmann glias, while an open arrowheads indicate an astrocytes in the granular layer. Both of them expressed Venus moderately. R, Confocal photomicrograph of Venus expression in Bergmann glias in a mostly Aldoc-negative area. Lobule V in a parasagittal section. Asterisks indicate PCs. Arrowheads indicate vertical processes of Bergmann glias. Scale bar in K applies to D–K. Scale bar in P applies to L–P. Scale bar in R applies to Q and R. See the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a> for abbreviations.</p

Inter-individual variation in Venus expression pattern in the cerebellum.

<p>A–F, Photomicrographs of vermal lobules VII–VIII in five adult mice (A–E) and a schematic representation of the striped expression pattern of Venus in this area (F). G–L, Photomicrographs of the right side of lobules VI and its hemispheral extension, simple lobule and crus I in five adult mice (G–K) and a schematic representation of the striped expression pattern of Venus in this area (L). For abbreviations of anatomical terms, see the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a>. Arrowhead in J indicates a trace of the blood vessel. Scale bar in E applies to A–F. Scale bar in K applies to G–L.</p

Relationship of the 9+/9− boundary in the flocculus with the known floccular compartmentalization.

<p>A, Comparison of the Venus expression pattern (left), the immunostained HSP25 expression pattern (center) and double labeling (right) in photomicrographs of a horizontal section (top), SSAA from serial horizontal sections (middle), and mapping in the unfolded scheme (bottom), of the paraflocculus and flocculus. Arrowheads in the top panels indicate the boundary between Aldoc-positive and -negative areas, which coincided with the boundary between HSP25-positive and -negative areas, in the flocculus. Arrowheads in the center panels indicate the apex of the lobule. The displays of SSAA were shrunk by 50% in the dorsoventral (vertical) direction. The mapping was based on SSAA. B, Three dimensional reconstruction of the HSP25 and Venus labeling patterns in the paraflocculus and flocculus. Caudal and rostral views of the paraflocculus and a lateral view of the flocculus are shown. Red and pink areas indicate strong and moderate HSP25 expression. Purple areas indicate Aldoc-negative area. C, Schematic showing correspondence between the Aldoc stripes and functional zones in the flocculus. D, Anterograde labeling of olivocerebellar climbing fibers by a tracer injection into the dorsal cap subnucleus of the inferior olive. Comparison of labeled climbing fibers (center), Venus expression (left) and double labeling (right) indicates that labeled climbing fibers were located in the Aldoc-negative area. E, Injection site of tracer (Alexa Fluor 594-conjugated dextran amine) in the dorsal cap subnucleus of the inferior olive. Contour of the inferior olive subnuclei (blue) was drawn by referring to another photo of intrinsic and Venus fluorescence of the same section. See the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a> for abbreviations.</p

Identification of Aldoc expressing neurons and glia in the retina with confocal photomicroscopy.

<p>A, Transverse section of the retina under low magnification. Only this photo was taken with an epifluorescence microscope. This retina section was obtained from a perfused mouse. B, Intermediate magnification photos of a cross section labeled with immunostaining of Venus with anti-GFP antibody (left subpanel), DAPI staining (center subpanel) and double labeling (right subpanel). C–H, High magnification photos of cross sections labeled with immunostaining of Venus (most left subpanel), immunostaining of calbindin (C), recoverin (D), Pax6 (E), cone arrestin (F), glutamine synthetase (G) or protein kinase C (H) (next most left subpanel), DAPI staining (next most right subpanel), and triple labeling (most right subpanel). Arrowheads indicate a population of ganglion cell (B), horizontal cell (C), rod photoreceptor cells (D), amacrine cells (E), cone photoreceptor cells (F), Müller glia cells (G) and bipolar cell (H). See the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a> for abbreviations.</p

Detailed Aldoc expression pattern in the cerebellar vermis and hemisphere. examined in Aldoc-Venus mice.

<p>A–E, Aspects of vermal surface. F, Reconstruction of the unfolded entire vermal cortex (molecular layer) by SSAA from serial sections. The rostral and caudal parts (lobules I–V and lobules VII–X) were from serial coronal sections in a mouse and the central part (lobule V–VII) were from serial horizontal sections in another mouse. G–I, Aspects of hemispheral surface. J, Reconstruction of the unfolded entire hemispheral cortex (molecular layer) by SSAA from serial sections. Since folial organization is complicated, SSAAs in individual folia are combined to show the entire cortex as a mosaic of the SSAAs. K, The revised scheme of the Aldoc expression pattern drawn on the unfolded cerebellar cortex based on the SSAAs shown in F and J. Note that darker colors mean more intense expression of Venus in K, opposite to photomicrographs. The mosaic displays of SSAAs were shrunk by 45% in the rostrocaudal (vertical) direction to fit to the space in F and J. Arrowheads indicate the apex of lobules in F and J. Scale bar in E applies to A–E. Scale bar in I applies to G–I. See the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a> for abbreviations.</p

Comparison of the striped Aldoc or Venus expression pattern among wild type (<i>Aldoc</i>^+/+), heterozygote (<i>Aldoc</i>^+/Venus) and homozygote (<i>Aldoc</i>^Venus/Venus) mice.

<p>A–F, Photomicrographs of Aldoc immunostaining in a wild type (the most left column), Aldoc immunostaining, double labeling and Venus in a heterozygote (the three center columns), and Venus in a homozygote (the most right column) in horizontal sections of the cerebella at the similar levels (A, rostral cerebellum with three narrow zones; B, Paravermal lobule VIa; C, Lobule VIII; D, Junction between lobules IXc and X; E, Flocculus, dorsal part; F, Flocculus, ventral part). Arrowheads in D indicate areas with higher Aldoc/Venus expression in lobule X. White and black arrowheads in E and F indicate rostral and caudal parts of the flocculus that had different Aldoc/Venus expression levels. Sections from the wild type and heterozygote were processed with Aldoc immunostaining with Texas red-conjugated secondary antibody in the same session with the same antibody solutions. Photos of Aldoc immunostaining and Venus were, respectively, processed with the same exposure and adjustment settings for all mice in A–F. Scale bar under F applies to A–F. G, Higher magnification photomicrographs of Aldoc immunostaining (left panel), double labeling (center panel) and Venus (right panel) of a coronal section of the cerebellum (lobule VIII) in a heterozygote, showing complete match at the cell population level.</p

Venus expression pattern in the flocculus and paraflocculus.

<p>A, Photomicrograph of the flocculus in the ventral outer surface of the whole-mount preparation of the right cerebellum. Venus expression in the rostral part was weaker than in the caudal part. B–D, Photomicrograph of double labeling of PLCB4 in a parasagittal section of the flocculus. Superimposing (D) of immunostaining of PLCB4 (B) and Venus fluorescence (C) in the same section indicate that PLCB4 and Venus are expressed in a complementary pattern; the rostral part that was weak in Venus expression expressed PLCB4. Arrowheads indicate the boundary between the Venus-positive and -negative areas. Inset in the right bottom indicates the position of the photomicrograph. E, Schematic illustrating lobular organization and unfolding of the flocculus. F, The Aldoc expression pattern in the paraflocculus and flocculus depicted in the unfolded scheme, based on the SSAA (G). G, SSAA from serial parasagittal sections of the paraflocculus and flocculus. The displays of SSAA were shrunk by 50% in the dorsoventral (vertical) direction. Red curve indicate the boundary between stripes 8a+ and 8b+. Arrowheads indicate the apex of lobules. Scale bar in D applies to B–D. See the legends for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0086679#pone-0086679-g001" target="_blank">Figure 1</a> for abbreviations.</p

Radioactivity versus time curves in the brains of MPTP-free and MPTP-treated marmosets.

<p>A dopamine transporter ligand, [¹¹C]PE2I, was intravenously administered to marmosets. The putamen and caudate in the striatum were the target regions, and the cerebellum was a reference region.</p

Representative parametric images.

<p>Coronal sections illustrating the binding potential (BP_ND) of [¹¹C]PE2I in the brains of MPTP-free and MPTP-treated marmosets are presented.</p

Binding potentials (BP_ND) of [¹¹C]PE2I.

<p>Binding potentials in the striatum (putamen and caudate) of brains of MPTP-treated and MPTP-free common marmosets are presented.</p>#1<p>:Ratios of mean daily locomotion counts of the post-MPTP period to those of pre-MPTP period (mean ± SD).</p>#2<p>:MPTP cumulative doses (mg/kg).</p>*<p>:p<0.05 against MPTP-free marmosets (Bonferroni test). SD: Standard deviation.</p