Journal: Cerebellum (London, England)

Article Title: Climbing fiber development is impaired in postnatal Car8 wdl mice

doi: 10.1007/s12311-017-0886-1

Figure Lengend Snippet: (a) Schematics of the olivo-cerebellar-nuclei circuit (left) and CF terminals on a PC (right). Abbreviations: IHC = immunohistochemistry, ml = molecular layer, pcl = Purkinje cell layer (white dotted lines), gcl = granule cell layer, cn = cerebellar nuclei, IO = inferior olive. (b) VGLUT2 and calbindin double staining reveals premature CF elimination and delayed CF translocation into the molecular layer at P10 (scale bar = 50 μm). The number (no) of VGLUT2-positive puncta on PC somata (p<0.0001) and percent CF territory (p=0.0159) were significantly different between Car8wdl and control mice at P10. In the graphs, * = 0.0159, and **** < 0.0001. Error bars reflect ± the standard of the mean (SEM). Yellow arrowheads point to the CF terminals that have translocated from the PC somata onto the PC dendrites. (c) Higher magnification of VGLUT2-positive CF terminals on PC somata (white arrowheads) and dendrites (yellow arrowheads) at P10 (scale bar = 20 μm). Control mice have more VGLUT2-positive CF terminals on PC somata and dendrites compared to the Car8wdl mutant mice.

Article Snippet: We used either anti-rabbit calbindin polyclonal (Swant #CB38) or anti-mouse calbindin monoclonal antibodies (Swant #300) both at a 1:10,000 dilution to label PCs.

Techniques: Immunohistochemistry, Double Staining, Translocation Assay, Control, Mutagenesis

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Structural Correlates of Efficient GABAergic Transmission in the Basal Ganglia-Thalamus Pathway

doi: 10.1523/JNEUROSCI.5266-07.2008

Figure Lengend Snippet: Topography of nigrothalamic terminals and their relationship to calbindin immunostaining in the rat. A, Schematic labeling of five injection sites in the SNR shown at five coronal levels of the atlas of Paxinos and Watson (1998). B-G, Distribution of the dense core of nigrothalamic terminal fields in the thalamus, color coded according to the labeling in A. Scattered fibers are omitted for clarity. The outlines of the terminal fields are shown overlaid on calbindin-immunostained coronal sections of the thalamus sampled 300 μm apart. Note the dual terminal fields in the ventromedial and intralaminar nuclei and the long anteroposterior extent of the terminal fields. Caudal SNR injections result in more dense intralaminar fiber plexuses, whereas rostral injections label only few fibers in these nuclei. H, I, Light microscopic images of the VM double stained for BDA (black) and calbindin (brown) after injection of the tracer into SNR. The boxed area in H is shown enlarged in I. Nigrothalamic terminals (arrows in I) are restricted to the zone containing calbindin-immunoreactive cells (asterisks). J, Nigrothalamic terminals establish multiple contacts (arrows) on the soma of a calbindin-positive relay cell. I and J are multifocal images. Scalebars:B-G, 1 mm; H, 200 μm; I, 50 μm; J, 10 μm. AV, Anteroventral thalamic nucleus (n.); CL, centrolateral thalamic n.; F, nucleus of the fields of Forel; fr, fasciculus retroflexus; Hb, habenular n.; LD, laterodorsal thalamic n.; LH, lateral hypothalamic area; LP, lateral posterior thalamic n.; MD, mediodorsal thalamic n.; ml, medial lemniscus; mt, mammillothalamic tract; PaR, pararubral nucleus; PC, paracentral thalamic n.; PR, prerubral field; Po, posterior thalamic n. group; RMC, red nucleus, magnocellular part; RPC, red nucleus, parvicellular part; sm, stria medullaris of thalamus; SNC, substantia nigra, compact part; Sub, submedius thalamic n.; VA, ventral anterior thalamic n.; VL, ventrolateral thalamic n.; VTA, ventral tegmental area; ZID, zona incerta dorsal part; ZIV, zona incerta ventral part.

Article Snippet: After visualizing the tracer with DABNi, sections containing anterogradely labeled terminals were incubated with mouse anti-calbindin antiserum (1:4000, overnight; Swant, Bellinzona, Switzerland).

Techniques: Immunostaining, Labeling, Injection, Staining

Journal: Journal of Cell Science

Article Title: A polycystin-2 protein with modified channel properties leads to an increased diameter of renal tubules and to renal cysts

doi: 10.1242/jcs.259013

Figure Lengend Snippet: Polycystic kidney disease in Pkd2 poreL1 knock-in mice. (A,B) Transverse kidney sections from 12-month-old wild-type (+/+) and homozygous Pkd2 poreL1 knock-in (p/p) were stained with H&E. Whereas kidney cysts can be easily seen at the age of 12 months on a 129/Sv background, no kidney cysts were observed on a C57Bl/6 background over a period of 12 months. Scale bars: 1 mm. (C,D) Immunohistochemical staining of kidney sections from homozygous Pkd2 poreL1 knock-in mice on a 129/Sv background for aquaporin-2 (a marker of principal cells in collecting ducts) shows that cysts originate in collecting ducts (C). Double-immunofluorescence staining for aquaporin-2 (green) and calbindin (a marker of connecting tubules, red) demonstrates the sudden transition from a calbindin-positive connecting tubule to an aquaporin-2-positive collecting duct cyst (asterisk in D). Scale bars: 100 µm. (E–H) Scanning electron micrographs were taken from kidneys of 3-, 6- and 12-month-old wild-type and Pkd2 poreL1 knock-in mice of both sexes on a 129/Sv background to determine the length of primary cilia. No obvious morphological alterations of primary cilia in collecting ducts were detected (E). Scale bars: 2 µm. At all three ages the length of primary cilia in the knock-in mice is shifted towards higher values. The number of cilia analyzed is given in the upper right-hand corner of the line graphs in panels F–H. Images are representative of at least three animals per condition.

Article Snippet: The following primary antibodies were used: a mouse monoclonal anti-calbindin antibody (Merck, cat. no. C8666; diluted 1:4000), a polyclonal goat anti-aquaporin-2 antibody (Santa Cruz Biotechnology, cat. no. sc-9882; diluted 1:200), a rabbit polyclonal anti-laminin antibody (Merck, cat. no. L9393; diluted 1:100), a polyclonal rabbit anti-Na + /Cl − cotransporter antiserum (diluted 1:1000) , a polyclonal rabbit anti-uromodulin antibody (Biotrend, cat. no. 8595-0004; diluted 1:100), and the polyclonal rabbit anti-polycystin-2 antibodies YCB9 and YCC2 (diluted 1:200) ( ).

Techniques: Knock-In, Staining, Immunohistochemical staining, Marker, Double Immunofluorescence Staining