Spontaneous postsynaptic currents in Purkinje cells.

<p><b>(A)</b> Representative traces of spontaneous EPSCs and IPSCs in control and <i>Cstb^−/−</i> Purkinje cells. EPSCs are seen as inward currents (downward deflections) and IPSCs are seen as outward currents (upward deflections). Insets show single IPSCs and EPSCs taken from the time points indicated by asterisks. An arrow indicates a synchronous burst of IPSCs seen in control but not in <i>Cstb^−/−</i> Purkinje cells. <b>(B)</b> The occurrence of IPSCs and synchronous burst of IPSCs on control and <i>Cstb^−/−</i> Purkinje cells. Individual cells measured are shown as spheres. <b>(C)</b> The frequency of EPSCs was significantly higher in <i>Cstb^−/−</i> cells compared to controls (p = 0.034). The data are expressed as mean frequency (Hz) ± SE. *, p<0.05; **, p<0.01.</p

The GO terms of biological processes, molecular functions and cellular components in P30 <i>Cstb^−/−</i> cerebellum.

<p>The GO terms of biological processes, molecular functions and cellular components in P30 <i>Cstb^−/−</i> cerebellum.</p

The GO terms of biological processes, molecular functions and cellular components in P7 <i>Cstb^−/−</i> cerebellum.

<p>The GO terms of biological processes, molecular functions and cellular components in P7 <i>Cstb^−/−</i> cerebellum.</p

Autoradiography of GABA_A receptors.

<p><b>(A)</b> Binding of [³H]muscimol was significantly reduced in P30 <i>Cstb^−/−</i> cerebellum (p = 0.037). No change was seen at P7. <b>(B)</b> Binding of [³H]Ro15-4513 did not show significant changes at P7 or at P30. However, when diazepam (DZ) was added to reveal the diazepam-insensitive α6 subunit-dependent GABA_A receptor subtype, decreased binding for [³H]Ro15-4513 (p = 0.012) was seen. The data are expressed as mean radioactivity levels (nCi/mg) ± SE. *, p<0.05. <b>(C)</b> Representative images of [³H]muscimol binding to P30 <i>Cstb^−/−</i> and control brain. <b>(D)</b> Representative images of [³H]Ro15-4513 binding to P30 <i>Cstb^−/−</i> and control brain without and <b>(E)</b> in presence of diazepam.</p

The expression of synapsin 1, VGAT and gephyrin positive synaptic puncta.

<p><b>(A)</b> The number of synapsin 1 positive puncta in the molecular layer of <i>Cstb^−/−</i> cerebellum was significantly lower compared to controls at P14 (p = 0.0035) and P20 (p = 0.0447). P7 and P30 animals did not show significant difference. <b>(B)</b> Molecular layer of P14 and P20 <i>Cstb^−/−</i> cerebellum shows less synapsin 1 positive puncta compared to control. <b>(C)</b> The number of VGAT positive puncta in the molecular layer of <i>Cstb^−/−</i> cerebellum was significantly lower compared to controls at P14 (p = 0.0115). P7, P20 and P30 animals did not show significant difference. <b>(D)</b> Molecular layer of P14 <i>Cstb^−/−</i> cerebellum shows less VGAT positive puncta compared to control. <b>(E)</b> The number of gephyrin positive puncta in the molecular layer of <i>Cstb^−/−</i> cerebellum was significantly lower compared to controls at P20 (p = 0.0448) and P30 (p = 0.0131). P7 and P14 animals did not show significant difference. <b>(F)</b> Molecular layer of P20 and P30 <i>Cstb^−/−</i> cerebellum shows less gephyrin positive puncta compared to control. The data are expressed as mean amount of positive puncta relative to control ± SE. *, p<0.05; **, p<0.01.</p

Differential expression of <i>Gabra6</i> and <i>Gabrd</i>.

<p>qPCR shows increased expression of <i>Gabra6</i> and <i>Gabrd</i> in P7 <i>Cstb^−/−</i> cerebellum compared to the control mice. The data are expressed as a fold change relative to controls ± SE. *, p<0.05.</p

The GO terms of biological processes, molecular functions and cellular components in P5+2 DIV <i>Cstb^−/−</i> cerebellar granule cells.

<p>The GO terms of biological processes, molecular functions and cellular components in P5+2 DIV <i>Cstb^−/−</i> cerebellar granule cells.</p

The colocalization of PAP (green) and GAD65/67 (red) was seen in several areas of brain.

<p>In cerebral Purkinje cells (A–C), strong colocalization was seen especially in the axon hillock of the neuron (small picture in C; yellow color and white arrows depicting the colocalization). Similarly, PAP was present in GABAergic neurons in prefrontal cortex (PFC; infralimbic cortex) (D–F). Scale bars are 10 µm.</p

Immunofluorescent colocalization stainings of TMPAP and synaptic vesicle associated proteins.

<p>TMPAP (green) is colocalized with a presynaptic marker, synaptophysin (red) (A–C; yellow color and white arrows depicting the colocalization). PAP was seen in vesicle-like structures that had strong colocalization with Snapin (D–F). Small picture is a magnification from panel C, depicting the colocalization. Moreover, largest PAP-immunoreactive structures had a colocalization with multivesicular bodies (MVB, red; G–I). All pictures are from striatum. Scale bars are 10 µm in A–C and G–I, and 3 µm in D–F.</p

TMPAP is expressed in the mouse brain and colocalizes with GABAergic marker, GAD 65/67.

<p>Representative confocal images depict intense TMPAP (brown color) expression in molecular cell layer (M) and Purkinje cells (P) of cerebellum (Panel A), in substantia nigra pars reticulata (SNpr; Panel B), in red nucleus (RN; Panel C) and in oculomotor nucleus (O; Panel C). Small picture in Panel B depicts the TMPAP staining of the substantia nigra in PAP<i>^−/−</i> mouse. TMPAP (green) was colocalized with GABAergic marker (red) in medium spiny neurons of striatum (Panels D-G, yellow color and white arrows indicating the colocalization) and in SNpr (Panels H–K). Colocalization was evident also in GABAergic neurons of hippocampus CA1 (Panels L–O). DAPI (blue color) was used as a nuclear marker. Scale bars are 500 µm in Panels A–C, and 10 µm in panels D–O.</p