Journal: Human Molecular Genetics

Article Title: Viral vector-mediated SLC9A6 gene replacement reduces cerebellar motor and molecular abnormalities in the shaker rat model of Christianson syndrome

doi: 10.1093/hmg/ddag021

Figure Lengend Snippet: AAV9-CAG-hSLC9A6 AAV increased cerebellar NHE6 and CALB1 abundances determined by western blotting and decreased motor ataxia. (A-D) Mirroring findings with PHP.eB-L7-Slc9a6-GFP AAV, administration of AAV9-CAG-hSLC9A6 AAV at ~ 5 weeks of age minimally increased NHE6 expression (A, C), but this minimal increase led to proportionally greater increases in CALB1 expression (B, D). (E) AAV9-CAG-hSLC9A6 AAV administration at 5 weeks of age significantly improved motor ataxia at 14 weeks of age compared to 7 untreated shaker rats (5 administered a control AAV and 2 uninjected). (F) Rats treated with AAV9-CAG-hSLC9A6 trended towards a decrease in tremor at 14 weeks of age, albeit with substantial variability. Note that each data point is an individual rat, and all n-values for statistics are calculated as number of rats, not technical replicates.

Article Snippet: To generate hSLC9A6 AAV expression plasmid, pAAV-CAG-GFP plasmid (Addgene plasmid #37825, gifted from Edward Boyden) was modified by deleting GFP with BamHI and EcoRI restriction enzymes.

Techniques: Western Blot, Expressing, Control

Journal: Human Molecular Genetics

Article Title: Viral vector-mediated SLC9A6 gene replacement reduces cerebellar motor and molecular abnormalities in the shaker rat model of Christianson syndrome

doi: 10.1093/hmg/ddag021

Figure Lengend Snippet: Following AAV9-CAG-hSLC9A6 AAV administration, ataxia, NHE6, and CALB1, are interrelated, but NHE6 and CALB1 are not correlated with tremor at 14 weeks of age. (A) NHE6 abundance following viral administration of AAV9-CAG-hSLC9A6 AAV positively correlates with CALB1 expression, suggesting a rescue of Purkinje cells. (B) NHE6 abundance is likewise negatively correlated with gait ataxia. (C) CALB1 is highly negatively correlated with gait ataxia. (D-E) While NHE6 and CALB1 abundance were correlated with ataxia and ataxia is correlated with tremor, NHE6 and CALB1 are not significantly correlated with tremor. Grey shaded regions represents 95% confidence intervals.

Article Snippet: To generate hSLC9A6 AAV expression plasmid, pAAV-CAG-GFP plasmid (Addgene plasmid #37825, gifted from Edward Boyden) was modified by deleting GFP with BamHI and EcoRI restriction enzymes.

Techniques: Expressing

Journal: Human Molecular Genetics

Article Title: Viral vector-mediated SLC9A6 gene replacement reduces cerebellar motor and molecular abnormalities in the shaker rat model of Christianson syndrome

doi: 10.1093/hmg/ddag021

Figure Lengend Snippet: (A) In the context of AAV9-CAG-hSLC9A6 AAV experiments, ataxia is strongly correlated with tremor at 14 weeks. (B-C) In the context of PHP.eB-Slc9a6-GFP AAV experiments, the correlation between peak tremor and late-stage ataxia is stronger than the correlation between tremor and ataxia at any given timepoint. (B) Ataxia and tremor are initially strongly related following symptom onset, but this correlation significantly decreases over time. (C) Peak tremor occurred prior to 20 weeks in all but 2 animals; however, the correlation between peak ataxia and late-stage ataxia (21–25 weeks) was higher than the correlation between tremor and ataxia at any individual timepoint. (D) Endpoint NHE6 expression and ataxia were negatively correlated at all timepoints. This negative correlation became stronger over time. (E) NHE6 and tremor were negatively correlated at all timepoints; however, the correlation between NHE6 and tremor did not strengthen over time. Grey shaded regions represents 95% confidence intervals.

Article Snippet: To generate hSLC9A6 AAV expression plasmid, pAAV-CAG-GFP plasmid (Addgene plasmid #37825, gifted from Edward Boyden) was modified by deleting GFP with BamHI and EcoRI restriction enzymes.

Techniques: Expressing