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Scavenging <t>endogenous</t> <t>BDNF</t> prevents the astrocyte-induced increase in the number and synaptic localization of GABAAR clusters. Hippocampal cultures with and without astrocytes were treated with 2.0 μg/ml <t>TrkB-IgG</t> to scavenge endogenous BDNF or with TrkC-IgG to scavenge NT3 at different ages in vitro. Immunostaining was performed with antibodies against GABAAR-β2/β3 (green) and SP (red) at 4, 7, or 10 div. A, Treatment with 2.0 μg/ml TrkB-IgG beginning at 1 div resulted in fewer postsynaptic GABAAR clusters in pure neuronal cultures (top) and neuron-astrocyte cocultures (bottom) at 4, 7, and 10 div. No change was observed in the number of SP+ boutons after BDNF scavenging. Scale bar, 2 μm. B, Quantification of TrkB-IgG effects on the number of GABAAR clusters per 20 μm dendrite segment. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. C, Quantification of TrkB-IgG effects on the synaptic localization of GABAAR clusters. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. D, Treatment with 2.0 μg/ml TrkB-IgG at 8-10 div for 48 h decreased the number of postsynaptic GABAAR clusters in neuron-astrocyte cocultures (middle) (Table 2) compared with untreated controls (left). In contrast, treatment with 2.0 μg/ml TrkC-IgG for 48 h increased the proportion of GABAAR clusters localized to synapses but had no effect on the number of GABAAR clusters (right) (Table 2). No change was observed in the number of SP+ boutons after BDNF or NT3 scavenging. Scale bar, 2 μm. Error bars represent SEM.
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Scavenging <t>endogenous</t> <t>BDNF</t> prevents the astrocyte-induced increase in the number and synaptic localization of GABAAR clusters. Hippocampal cultures with and without astrocytes were treated with 2.0 μg/ml <t>TrkB-IgG</t> to scavenge endogenous BDNF or with TrkC-IgG to scavenge NT3 at different ages in vitro. Immunostaining was performed with antibodies against GABAAR-β2/β3 (green) and SP (red) at 4, 7, or 10 div. A, Treatment with 2.0 μg/ml TrkB-IgG beginning at 1 div resulted in fewer postsynaptic GABAAR clusters in pure neuronal cultures (top) and neuron-astrocyte cocultures (bottom) at 4, 7, and 10 div. No change was observed in the number of SP+ boutons after BDNF scavenging. Scale bar, 2 μm. B, Quantification of TrkB-IgG effects on the number of GABAAR clusters per 20 μm dendrite segment. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. C, Quantification of TrkB-IgG effects on the synaptic localization of GABAAR clusters. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. D, Treatment with 2.0 μg/ml TrkB-IgG at 8-10 div for 48 h decreased the number of postsynaptic GABAAR clusters in neuron-astrocyte cocultures (middle) (Table 2) compared with untreated controls (left). In contrast, treatment with 2.0 μg/ml TrkC-IgG for 48 h increased the proportion of GABAAR clusters localized to synapses but had no effect on the number of GABAAR clusters (right) (Table 2). No change was observed in the number of SP+ boutons after BDNF or NT3 scavenging. Scale bar, 2 μm. Error bars represent SEM.
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Scavenging endogenous BDNF prevents the astrocyte-induced increase in the number and synaptic localization of GABAAR clusters. Hippocampal cultures with and without astrocytes were treated with 2.0 μg/ml TrkB-IgG to scavenge endogenous BDNF or with TrkC-IgG to scavenge NT3 at different ages in vitro. Immunostaining was performed with antibodies against GABAAR-β2/β3 (green) and SP (red) at 4, 7, or 10 div. A, Treatment with 2.0 μg/ml TrkB-IgG beginning at 1 div resulted in fewer postsynaptic GABAAR clusters in pure neuronal cultures (top) and neuron-astrocyte cocultures (bottom) at 4, 7, and 10 div. No change was observed in the number of SP+ boutons after BDNF scavenging. Scale bar, 2 μm. B, Quantification of TrkB-IgG effects on the number of GABAAR clusters per 20 μm dendrite segment. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. C, Quantification of TrkB-IgG effects on the synaptic localization of GABAAR clusters. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. D, Treatment with 2.0 μg/ml TrkB-IgG at 8-10 div for 48 h decreased the number of postsynaptic GABAAR clusters in neuron-astrocyte cocultures (middle) (Table 2) compared with untreated controls (left). In contrast, treatment with 2.0 μg/ml TrkC-IgG for 48 h increased the proportion of GABAAR clusters localized to synapses but had no effect on the number of GABAAR clusters (right) (Table 2). No change was observed in the number of SP+ boutons after BDNF or NT3 scavenging. Scale bar, 2 μm. Error bars represent SEM.

Journal: The Journal of Neuroscience

Article Title: Astrocytes Regulate Inhibitory Synapse Formation via Trk-Mediated Modulation of Postsynaptic GABA A Receptors

doi: 10.1523/JNEUROSCI.3980-04.2005

Figure Lengend Snippet: Scavenging endogenous BDNF prevents the astrocyte-induced increase in the number and synaptic localization of GABAAR clusters. Hippocampal cultures with and without astrocytes were treated with 2.0 μg/ml TrkB-IgG to scavenge endogenous BDNF or with TrkC-IgG to scavenge NT3 at different ages in vitro. Immunostaining was performed with antibodies against GABAAR-β2/β3 (green) and SP (red) at 4, 7, or 10 div. A, Treatment with 2.0 μg/ml TrkB-IgG beginning at 1 div resulted in fewer postsynaptic GABAAR clusters in pure neuronal cultures (top) and neuron-astrocyte cocultures (bottom) at 4, 7, and 10 div. No change was observed in the number of SP+ boutons after BDNF scavenging. Scale bar, 2 μm. B, Quantification of TrkB-IgG effects on the number of GABAAR clusters per 20 μm dendrite segment. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. C, Quantification of TrkB-IgG effects on the synaptic localization of GABAAR clusters. *p < 0.001, significant difference compared with no treatment in pure neuronal cultures; **p < 0.001, significant decrease compared with no treatment in neuron astrocyte cocultures. D, Treatment with 2.0 μg/ml TrkB-IgG at 8-10 div for 48 h decreased the number of postsynaptic GABAAR clusters in neuron-astrocyte cocultures (middle) (Table 2) compared with untreated controls (left). In contrast, treatment with 2.0 μg/ml TrkC-IgG for 48 h increased the proportion of GABAAR clusters localized to synapses but had no effect on the number of GABAAR clusters (right) (Table 2). No change was observed in the number of SP+ boutons after BDNF or NT3 scavenging. Scale bar, 2 μm. Error bars represent SEM.

Article Snippet: To evaluate the role of BDNF and TrkB signaling, cultures were prepared from postnatal day 0 mice mutant for TrkB ( Klein et al., 1993 ) (The Jackson Laboratory, Bar Harbor, ME) or BDNF ( Ernfors et al., 1994 ) (The Jackson Laboratory) or from wild-type littermate controls.

Techniques: In Vitro, Immunostaining

Effects of neurotrophin manipulations on astrocyte modulation of postsynaptic GABA A R clusters

Journal: The Journal of Neuroscience

Article Title: Astrocytes Regulate Inhibitory Synapse Formation via Trk-Mediated Modulation of Postsynaptic GABA A Receptors

doi: 10.1523/JNEUROSCI.3980-04.2005

Figure Lengend Snippet: Effects of neurotrophin manipulations on astrocyte modulation of postsynaptic GABA A R clusters

Article Snippet: To evaluate the role of BDNF and TrkB signaling, cultures were prepared from postnatal day 0 mice mutant for TrkB ( Klein et al., 1993 ) (The Jackson Laboratory, Bar Harbor, ME) or BDNF ( Ernfors et al., 1994 ) (The Jackson Laboratory) or from wild-type littermate controls.

Techniques: