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Distribution of CB1 immunoreactivity in the binocular region of V1 at different postnatal ages. Mean and SEM of CB1 signal intensity in each layer represented as the proportion to the all-layer intensity (n = 4 animals, one-way factorial ANOVA, p,0.05; layer II/III, p.0.05; layers IV, V, and VI, post hoc Tukey’s test, *: p,0.05, **: p,0.01). doi:10.1371/journal.pone.0053082.gsecondary visual cortex (V2M), gradually decreased across cortical regions toward the V1 binocular region (BR) and increased again in the lateral area of the secondary visual cortex (V2L) (Fig. 1C, D). The signal intensity of V2M was significantly ��-Sitosterol ��-D-glucoside higher than that of BR in V1 (Fig. 1E).(Fig. 2E), suggesting that CB1 is mainly localized at the VGATpositive inhibitory nerve terminals in V1.Developmental Changes in CB1 Expression in VTo address the possible role of CB1 in the developmental plasticity of V1, we explored the developmental regulation of CB1 in V1. The relative amount of CB1 protein in V1 gradually increased during development from P10 to P100 (Fig. 3A, B). The relative amount of CB1 at P100 was significantly higher than that at P20 (Fig. 3B). In the mice from P20 to P100, intense CB1 immunoreactivity was mainly observed in layers II/III and VI, while intense immunoreactivity was observed in layers I and VI in P10 animals (Fig. 3C). In layer II/III, the CB1 immunoreactivity between P30 and P50 was significantly higher than that of P10 (Fig. 3D).Synaptic Localization of CB1 in VTo elucidate the synaptic localization of CB1, we performed double immunofluorescent staining of CB1 and MAP2 or synaptophysin in the V1 of P30 mice (Fig. 2A, B). An immunopositive CB1 signal was observed in the structures that consist of shafts and varicosities. In the upper layer of V1, CB1positive varicosities appeared to contact the soma and MAP2positive dendrites (Fig. 2A). To confirm the presynaptic characteristics of the CB1-positive varicosities, we evaluated the colocalization of CB1 and synaptophysin signals in the CB1 positive varicosities and shafts by calculating CC values (Fig. 2B). The CC value in the varicosities was significantly higher than that in the shafts (Fig. 2C), suggesting the presynaptic nature of CB1positive varicosities. CB1 is found in both excitatory and 24195657 inhibitory nerve terminals [11]. To determine the synaptic localization of CB1 in the V1 of P30 mice, we examined the colocalization of immunopositive signals of CB1 and VGluTs or VGAT. Representative double immunofluorescent staining of CB1 and VGluTs or VGAT is shown in Fig. 2D. We evaluated the colocalization of CB1 and the terminal markers by calculating the CC values in the CB1-positive varicosities. The CC values of CB1 and VGAT were significantly higher 15826876 than those of CB1 and VGluTs in all cortical layers370-86-5 effect of Dark Rearing on CB1 ExpressionTo explore the effect of visual inputs on the developmental regulation of CB1 expression, we examined CB1 expression in mice that were dark reared from birth to P30 or P50. The mice reared in the dark from birth to P30 had a lesser quantity of CB1 protein than the normal mice reared under normal light/dark conditions. However, the mice that were dark reared until P50 had similar amounts of CB1 protein as the normal mice (Fig. 4A, B). In P30 animals, the pattern of layer distribution of CB1 was similar between the dark-reared and normal groups (Fig. 4C, D). To determine the effect of dark rearing on the synaptic localization of CB1, we compared the colocaliz.Distribution of CB1 immunoreactivity in the binocular region of V1 at different postnatal ages. Mean and SEM of CB1 signal intensity in each layer represented as the proportion to the all-layer intensity (n = 4 animals, one-way factorial ANOVA, p,0.05; layer II/III, p.0.05; layers IV, V, and VI, post hoc Tukey’s test, *: p,0.05, **: p,0.01). doi:10.1371/journal.pone.0053082.gsecondary visual cortex (V2M), gradually decreased across cortical regions toward the V1 binocular region (BR) and increased again in the lateral area of the secondary visual cortex (V2L) (Fig. 1C, D). The signal intensity of V2M was significantly higher than that of BR in V1 (Fig. 1E).(Fig. 2E), suggesting that CB1 is mainly localized at the VGATpositive inhibitory nerve terminals in V1.Developmental Changes in CB1 Expression in VTo address the possible role of CB1 in the developmental plasticity of V1, we explored the developmental regulation of CB1 in V1. The relative amount of CB1 protein in V1 gradually increased during development from P10 to P100 (Fig. 3A, B). The relative amount of CB1 at P100 was significantly higher than that at P20 (Fig. 3B). In the mice from P20 to P100, intense CB1 immunoreactivity was mainly observed in layers II/III and VI, while intense immunoreactivity was observed in layers I and VI in P10 animals (Fig. 3C). In layer II/III, the CB1 immunoreactivity between P30 and P50 was significantly higher than that of P10 (Fig. 3D).Synaptic Localization of CB1 in VTo elucidate the synaptic localization of CB1, we performed double immunofluorescent staining of CB1 and MAP2 or synaptophysin in the V1 of P30 mice (Fig. 2A, B). An immunopositive CB1 signal was observed in the structures that consist of shafts and varicosities. In the upper layer of V1, CB1positive varicosities appeared to contact the soma and MAP2positive dendrites (Fig. 2A). To confirm the presynaptic characteristics of the CB1-positive varicosities, we evaluated the colocalization of CB1 and synaptophysin signals in the CB1 positive varicosities and shafts by calculating CC values (Fig. 2B). The CC value in the varicosities was significantly higher than that in the shafts (Fig. 2C), suggesting the presynaptic nature of CB1positive varicosities. CB1 is found in both excitatory and 24195657 inhibitory nerve terminals [11]. To determine the synaptic localization of CB1 in the V1 of P30 mice, we examined the colocalization of immunopositive signals of CB1 and VGluTs or VGAT. Representative double immunofluorescent staining of CB1 and VGluTs or VGAT is shown in Fig. 2D. We evaluated the colocalization of CB1 and the terminal markers by calculating the CC values in the CB1-positive varicosities. The CC values of CB1 and VGAT were significantly higher 15826876 than those of CB1 and VGluTs in all cortical layersEffect of Dark Rearing on CB1 ExpressionTo explore the effect of visual inputs on the developmental regulation of CB1 expression, we examined CB1 expression in mice that were dark reared from birth to P30 or P50. The mice reared in the dark from birth to P30 had a lesser quantity of CB1 protein than the normal mice reared under normal light/dark conditions. However, the mice that were dark reared until P50 had similar amounts of CB1 protein as the normal mice (Fig. 4A, B). In P30 animals, the pattern of layer distribution of CB1 was similar between the dark-reared and normal groups (Fig. 4C, D). To determine the effect of dark rearing on the synaptic localization of CB1, we compared the colocaliz.

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Author: Cholesterol Absorption Inhibitors