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Increased dendritic spine density in response to <t>alpha-Synuclein</t> overexpression occurs in pSer129-negative neurons within the injection location. A Mice expressing Green fluorescent protein (GFP) in Layer V neurons (Thy1-GFP) mice were injected with control vector <t>(AAV-mCherry)</t> in one hemisphere and vector <t>for</t> <t>α-Syn</t> overexpression (OE): <t>(AAV-α-Syn-IRES-mCherry)</t> in the contralateral hemisphere for within animal comparison. B Spines from basilar dendrites 6 weeks post-injection were quantified via confocal microscopy. Sections were immunostained with antibodies against GFP and pSer129-α-Syn and dendritic spines were counted. C Basilar dendrites were analyzed in bins based on distance from the soma and grouped according to the hemisphere treatment (control or α-Syn OE). For the α-Syn OE hemisphere, cells were further divided into pSer129-α-Syn positive or negative. pSer129-α-Syn-negative dendrites had greater spine density than both mCherry-negative dendrites on the control hemisphere and pSer129-α-Syn-positive dendrites from the same hemisphere (Mixed-effects model, Main effect: treatment: F(2,63) = 3.397, p = 0.0397). Individual comparisons were corrected via Tukey: * p = 0.0106, *** p = 0.0010
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Increased dendritic spine density in response to <t>alpha-Synuclein</t> overexpression occurs in pSer129-negative neurons within the injection location. A Mice expressing Green fluorescent protein (GFP) in Layer V neurons (Thy1-GFP) mice were injected with control vector <t>(AAV-mCherry)</t> in one hemisphere and vector <t>for</t> <t>α-Syn</t> overexpression (OE): <t>(AAV-α-Syn-IRES-mCherry)</t> in the contralateral hemisphere for within animal comparison. B Spines from basilar dendrites 6 weeks post-injection were quantified via confocal microscopy. Sections were immunostained with antibodies against GFP and pSer129-α-Syn and dendritic spines were counted. C Basilar dendrites were analyzed in bins based on distance from the soma and grouped according to the hemisphere treatment (control or α-Syn OE). For the α-Syn OE hemisphere, cells were further divided into pSer129-α-Syn positive or negative. pSer129-α-Syn-negative dendrites had greater spine density than both mCherry-negative dendrites on the control hemisphere and pSer129-α-Syn-positive dendrites from the same hemisphere (Mixed-effects model, Main effect: treatment: F(2,63) = 3.397, p = 0.0397). Individual comparisons were corrected via Tukey: * p = 0.0106, *** p = 0.0010
Wild Type α Synuclein Aav6 Cag Hsnca Wpre, supplied by Vector Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Increased dendritic spine density in response to <t>alpha-Synuclein</t> overexpression occurs in pSer129-negative neurons within the injection location. A Mice expressing Green fluorescent protein (GFP) in Layer V neurons (Thy1-GFP) mice were injected with control vector <t>(AAV-mCherry)</t> in one hemisphere and vector <t>for</t> <t>α-Syn</t> overexpression (OE): <t>(AAV-α-Syn-IRES-mCherry)</t> in the contralateral hemisphere for within animal comparison. B Spines from basilar dendrites 6 weeks post-injection were quantified via confocal microscopy. Sections were immunostained with antibodies against GFP and pSer129-α-Syn and dendritic spines were counted. C Basilar dendrites were analyzed in bins based on distance from the soma and grouped according to the hemisphere treatment (control or α-Syn OE). For the α-Syn OE hemisphere, cells were further divided into pSer129-α-Syn positive or negative. pSer129-α-Syn-negative dendrites had greater spine density than both mCherry-negative dendrites on the control hemisphere and pSer129-α-Syn-positive dendrites from the same hemisphere (Mixed-effects model, Main effect: treatment: F(2,63) = 3.397, p = 0.0397). Individual comparisons were corrected via Tukey: * p = 0.0106, *** p = 0.0010
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Increased dendritic spine density in response to alpha-Synuclein overexpression occurs in pSer129-negative neurons within the injection location. A Mice expressing Green fluorescent protein (GFP) in Layer V neurons (Thy1-GFP) mice were injected with control vector (AAV-mCherry) in one hemisphere and vector for α-Syn overexpression (OE): (AAV-α-Syn-IRES-mCherry) in the contralateral hemisphere for within animal comparison. B Spines from basilar dendrites 6 weeks post-injection were quantified via confocal microscopy. Sections were immunostained with antibodies against GFP and pSer129-α-Syn and dendritic spines were counted. C Basilar dendrites were analyzed in bins based on distance from the soma and grouped according to the hemisphere treatment (control or α-Syn OE). For the α-Syn OE hemisphere, cells were further divided into pSer129-α-Syn positive or negative. pSer129-α-Syn-negative dendrites had greater spine density than both mCherry-negative dendrites on the control hemisphere and pSer129-α-Syn-positive dendrites from the same hemisphere (Mixed-effects model, Main effect: treatment: F(2,63) = 3.397, p = 0.0397). Individual comparisons were corrected via Tukey: * p = 0.0106, *** p = 0.0010

Journal: Cellular and Molecular Neurobiology

Article Title: Enhanced Spine Stability and Survival Lead to Increases in Dendritic Spine Density as an Early Response to Local Alpha-Synuclein Overexpression in Mouse Prefrontal Cortex

doi: 10.1007/s10571-024-01472-7

Figure Lengend Snippet: Increased dendritic spine density in response to alpha-Synuclein overexpression occurs in pSer129-negative neurons within the injection location. A Mice expressing Green fluorescent protein (GFP) in Layer V neurons (Thy1-GFP) mice were injected with control vector (AAV-mCherry) in one hemisphere and vector for α-Syn overexpression (OE): (AAV-α-Syn-IRES-mCherry) in the contralateral hemisphere for within animal comparison. B Spines from basilar dendrites 6 weeks post-injection were quantified via confocal microscopy. Sections were immunostained with antibodies against GFP and pSer129-α-Syn and dendritic spines were counted. C Basilar dendrites were analyzed in bins based on distance from the soma and grouped according to the hemisphere treatment (control or α-Syn OE). For the α-Syn OE hemisphere, cells were further divided into pSer129-α-Syn positive or negative. pSer129-α-Syn-negative dendrites had greater spine density than both mCherry-negative dendrites on the control hemisphere and pSer129-α-Syn-positive dendrites from the same hemisphere (Mixed-effects model, Main effect: treatment: F(2,63) = 3.397, p = 0.0397). Individual comparisons were corrected via Tukey: * p = 0.0106, *** p = 0.0010

Article Snippet: The Thy1-GFP mice ( n = 4) received bilateral injections with control virus into one hemisphere and α-Synuclein overexpression into the other hemisphere: Control: AAV6-CAG-mCherry-WPRE (1.7 × 10 12 gc/mL, Vector Biolabs, 1 μL, left hemisphere) and overexpression: vector carrying cDNA coding for human, wild-type α-Synuclein: AAV6-CAG-hSNCA-IRES-mCherry-WPRE (1.0 × 10 12 gc/mL, VectorBiolabs, 1 μL right hemisphere).

Techniques: Over Expression, Injection, Expressing, Plasmid Preparation, Comparison, Confocal Microscopy