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A : 2PEF map of arterioles and venules (top) and of both vessel and GCaMP7f-expressing nNOS neurons (outlined in white, bottom). Vessels analyzed in B are circled in solid red or blue, and neurons are numbered (1,2,3) B : Arterioles, but not venules, dilate in response to whisker puffs, spontaneous whisking or motion (grooming). Representative Ca 2+ transients in neuron 1, 2, and 3 (middle). Overlay of normalized neural ensemble activity and arteriolar diameter changes highlighting the close correspondence between arteriolar and neural responses (bottom). C : Linear relationship between the magnitude of the ensemble activity and arteriolar dilatation in response to whisking or whisking and spontaneous motion. Notice that with whisking+motion the slope of the relationship is significantly greater than with whisking alone (multiple comparison of one-way analysis of <t>covariance</t> models). D : The distribution of correlation coefficients between arteriolar dilatation and nNOS Ca 2+ transients for individual neurons does not differ in the 3 mice studied (p>0.05; nested one-way ANOVA)(top). The strongest correlation is observed between ensemble activity and arteriolar dilatation (bottom). E : Representative arteriolar diameter changes relative to the timing of the peak Ca 2+ activity in individual neuron during 10 consecutive 5 sec whisker air puffs shown in B . Notice that in some neurons peak activity occur earlier than in other neurons. F : Average of ten raster plots in 3 mice showing early and late active nNOS neurons. Shading indicates standard deviation.
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aoctool statistical function of the matlab software  (MathWorks Inc)
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A : 2PEF map of arterioles and venules (top) and of both vessel and GCaMP7f-expressing nNOS neurons (outlined in white, bottom). Vessels analyzed in B are circled in solid red or blue, and neurons are numbered (1,2,3) B : Arterioles, but not venules, dilate in response to whisker puffs, spontaneous whisking or motion (grooming). Representative Ca 2+ transients in neuron 1, 2, and 3 (middle). Overlay of normalized neural ensemble activity and arteriolar diameter changes highlighting the close correspondence between arteriolar and neural responses (bottom). C : Linear relationship between the magnitude of the ensemble activity and arteriolar dilatation in response to whisking or whisking and spontaneous motion. Notice that with whisking+motion the slope of the relationship is significantly greater than with whisking alone (multiple comparison of one-way analysis of <t>covariance</t> models). D : The distribution of correlation coefficients between arteriolar dilatation and nNOS Ca 2+ transients for individual neurons does not differ in the 3 mice studied (p>0.05; nested one-way ANOVA)(top). The strongest correlation is observed between ensemble activity and arteriolar dilatation (bottom). E : Representative arteriolar diameter changes relative to the timing of the peak Ca 2+ activity in individual neuron during 10 consecutive 5 sec whisker air puffs shown in B . Notice that in some neurons peak activity occur earlier than in other neurons. F : Average of ten raster plots in 3 mice showing early and late active nNOS neurons. Shading indicates standard deviation.
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A : 2PEF map of arterioles and venules (top) and of both vessel and GCaMP7f-expressing nNOS neurons (outlined in white, bottom). Vessels analyzed in B are circled in solid red or blue, and neurons are numbered (1,2,3) B : Arterioles, but not venules, dilate in response to whisker puffs, spontaneous whisking or motion (grooming). Representative Ca 2+ transients in neuron 1, 2, and 3 (middle). Overlay of normalized neural ensemble activity and arteriolar diameter changes highlighting the close correspondence between arteriolar and neural responses (bottom). C : Linear relationship between the magnitude of the ensemble activity and arteriolar dilatation in response to whisking or whisking and spontaneous motion. Notice that with whisking+motion the slope of the relationship is significantly greater than with whisking alone (multiple comparison of one-way analysis of <t>covariance</t> models). D : The distribution of correlation coefficients between arteriolar dilatation and nNOS Ca 2+ transients for individual neurons does not differ in the 3 mice studied (p>0.05; nested one-way ANOVA)(top). The strongest correlation is observed between ensemble activity and arteriolar dilatation (bottom). E : Representative arteriolar diameter changes relative to the timing of the peak Ca 2+ activity in individual neuron during 10 consecutive 5 sec whisker air puffs shown in B . Notice that in some neurons peak activity occur earlier than in other neurons. F : Average of ten raster plots in 3 mice showing early and late active nNOS neurons. Shading indicates standard deviation.
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A : 2PEF map of arterioles and venules (top) and of both vessel and GCaMP7f-expressing nNOS neurons (outlined in white, bottom). Vessels analyzed in B are circled in solid red or blue, and neurons are numbered (1,2,3) B : Arterioles, but not venules, dilate in response to whisker puffs, spontaneous whisking or motion (grooming). Representative Ca 2+ transients in neuron 1, 2, and 3 (middle). Overlay of normalized neural ensemble activity and arteriolar diameter changes highlighting the close correspondence between arteriolar and neural responses (bottom). C : Linear relationship between the magnitude of the ensemble activity and arteriolar dilatation in response to whisking or whisking and spontaneous motion. Notice that with whisking+motion the slope of the relationship is significantly greater than with whisking alone (multiple comparison of one-way analysis of covariance models). D : The distribution of correlation coefficients between arteriolar dilatation and nNOS Ca 2+ transients for individual neurons does not differ in the 3 mice studied (p>0.05; nested one-way ANOVA)(top). The strongest correlation is observed between ensemble activity and arteriolar dilatation (bottom). E : Representative arteriolar diameter changes relative to the timing of the peak Ca 2+ activity in individual neuron during 10 consecutive 5 sec whisker air puffs shown in B . Notice that in some neurons peak activity occur earlier than in other neurons. F : Average of ten raster plots in 3 mice showing early and late active nNOS neurons. Shading indicates standard deviation.

Journal: bioRxiv

Article Title: Calcium transients in nNOS neurons underlie distinct phases of the neurovascular response to barrel cortex activation in awake mice

doi: 10.1101/2022.10.03.510654

Figure Lengend Snippet: A : 2PEF map of arterioles and venules (top) and of both vessel and GCaMP7f-expressing nNOS neurons (outlined in white, bottom). Vessels analyzed in B are circled in solid red or blue, and neurons are numbered (1,2,3) B : Arterioles, but not venules, dilate in response to whisker puffs, spontaneous whisking or motion (grooming). Representative Ca 2+ transients in neuron 1, 2, and 3 (middle). Overlay of normalized neural ensemble activity and arteriolar diameter changes highlighting the close correspondence between arteriolar and neural responses (bottom). C : Linear relationship between the magnitude of the ensemble activity and arteriolar dilatation in response to whisking or whisking and spontaneous motion. Notice that with whisking+motion the slope of the relationship is significantly greater than with whisking alone (multiple comparison of one-way analysis of covariance models). D : The distribution of correlation coefficients between arteriolar dilatation and nNOS Ca 2+ transients for individual neurons does not differ in the 3 mice studied (p>0.05; nested one-way ANOVA)(top). The strongest correlation is observed between ensemble activity and arteriolar dilatation (bottom). E : Representative arteriolar diameter changes relative to the timing of the peak Ca 2+ activity in individual neuron during 10 consecutive 5 sec whisker air puffs shown in B . Notice that in some neurons peak activity occur earlier than in other neurons. F : Average of ten raster plots in 3 mice showing early and late active nNOS neurons. Shading indicates standard deviation.

Article Snippet: To evaluate statistical differences between the linear correlations of two data sets, we generated one-way analysis of covariance models (Matlab function: aoctool) and ran multiple comparison test (Tukey-Kramer) of the group means (Matlab function: multcompare).

Techniques: Expressing, Whisker Assay, Activity Assay, Comparison, Standard Deviation