Laminar specificity and coverage of viral-mediated gene expression restricted to GABAergic interneurons and their parvalbumin subclass in marmoset primary visual cortex
Data files
Aug 29, 2024 version files 44.69 KB
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ManuscriptFig2_Data_Table_ver4.xlsx
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ManuscriptFig4_Data_Table_ver5.xlsx
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ManuscriptFig6_Data_Table_ver3.xlsx
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README.md
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Abstract
In the mammalian neocortex, inhibition is important for dynamically balancing excitation and shaping the response properties of cells and circuits. The various computational functions of inhibition are thought to be mediated by different inhibitory neuron types of which a large diversity exists in several species. Current understanding of the function and connectivity of distinct inhibitory neuron types has mainly derived from studies in transgenic mice. However, it is unknown whether knowledge gained from mouse studies applies to the non-human primate, the model system closest to humans. The lack of viral tools to selectively access inhibitory neuron types has been a major impediment to studying their function in the primate. Here, we have thoroughly validated and characterized several recently-developed viral vectors designed to restrict transgene expression to GABAergic cells or their parvalbumin (PV) subtype, and identified two types that show high specificity and efficiency in marmoset V1. We show that in marmoset V1 AAV-h56D induces transgene expression in GABAergic cells with up to 91-94% specificity and 79% efficiency, but this depends on viral serotype and cortical layer. AAV-PHP.eB-S5E2 induces transgene expression in PV cells across all cortical layers with up to 98% specificity and 86-90% efficiency, depending on layer. Thus, these viral vectors are promising tools for studying GABA and PV cell function and connectivity in the primate cortex.
README: Laminar specificity and coverage of viral-mediated gene expression restricted to GABAergic interneurons and their parvalbumin subclass in marmoset primary visual cortex
https://doi.org/10.5061/dryad.ht76hdrr3
Description of the data and file structure
Here, we have thoroughly validated and characterized several recently-developed viral vectors designed to restrict transgene expression to GABAergic cells or their parvalbumin (PV) subtype, and identified two types that show high specificity and efficiency in marmoset primary visual cortex.
ManuscriptFig2*Data*Table_ver4: This spreadsheet contains the values for the graphs found in Figure2. Null values are present for the Layer 4A and 4B mouse data taken from Xu et al. 2010 as this data was not published.
MeanPV% - Normalized percentage of parvalbumin positive cells found in each cortical layer.
MeanGABA% - Normalized percentage of GABA positive cells found in each cortical layer.
Mean PV cell density - Mean cell density of parvalbumin positive cells found in each cortical layer.
Mean GABA cell density - Mean cell density of GABA positive cells found in each cortical layer.
PV Mouse V1 Xu 2010 - Mean cell density of parvalbumin positive cells found in each cortical layer of the mouse as reported in Xu et al 2010.
GABA Mouse V1 Xu 2010 - Mean cell density of GABA positive cells found in each cortical layer of the mouse as reported in Xu et al 2010.
PV cells % GABA positive - Percent of parvalbumin positive cells that were also GABA positive.
GABA cells % PV positive - Percent of GABA positive cells that were also parvalbumin positive.
ManuscriptFig4*Data*Table_ver5: This spreadsheet contains the values for the graphs found in Figure4. The 3 null values in the Specificity data are due to low n's. The Standard Error of the Mean could not be computed for these cells.
% GABA IHC - Normalized percentage of GABA positive cells found in each cortical layer as determined by immunohistochemistry.
% AAV9 - Normalized percentage of AAV9 viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of AAV9 virally labeled cells.
% AAV7 - Normalized percentage of AAV7 viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of AAV7 virally labeled cells.
% AAV1 - Normalized percentage of AAV1 viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of AAV1 virally labeled cells.
ManuscriptFig6*Data*Table_ver3: This spreadsheet contains the values for the graphs found in Figure6. The null value in the Specificity data are due to low n. The Standard Error of the Mean could not be computed for this cell.
% PV IHC - Normalized percentage of PV positive cells found in each cortical layer immunohistochemistry.
% Small Volume Injection - Normalized percentage of AAV viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of virally labeled cells after small volume 100nL injections.
% Mid Volume Injection - Normalized percentage of AAV viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of AAV virally labeled cells after mid volume 180nL injections.
% Large Volume Injection - Normalized percentage of AAV1 viral infected cells found in each cortical layer (Fig 4A) as well as the specificity (Fig4B) and coverage (Fig4C) of AAV1 virally labeled cells after mid volume 180nL injections
Code/software
Microsoft Excel
Methods
To facilitate the application of these inhibitory-neuron specific viral vectors to studies of the primate cortex, we have performed a thorough validation and characterization of the laminar expression of reporter proteins mediated by several enhancer/promoter-specific AAVs. Here we report results from the two vector types that have shown the greatest specificity of transgene expression in marmoset primary visual cortex (V1); specifically, we have tested three serotypes of the h56D promoter-AAV that restricts gene expression to GABAergic neurons (Mehta et al., 2019), and one serotype of the S5E2 enhancer-AAV that restricts gene expression to PV cells (Vormstein-Schneider et al., 2020). Using injections of these viral vectors in marmoset V1, combined with immunohistochemical identification of GABA and PV neurons, we find that the laminar distribution of reporter protein expression mediated by the GABA- and PV-enhancer AAVs validated in this study resembles the laminar distribution of GABA-immunoreactive (GABA+) and PV-immunoreactive (PV+) cells, respectively, in marmoset V1