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The role of voltage-gated calcium channels in neurotransmitter phenotype specification: Coexpression and functional analysis in Xenopus laevis.

Lewis BB, Miller LE, Herbst WA, Saha MS - J. Comp. Neurol. (2014)

Bottom Line: Calcium activity was also analyzed on a single-cell level, and spike frequency was correlated with the expression of VGCC α1 subunits in cell culture.Cells expressing Cav 2.1 and Cav 2.2 displayed increased calcium spiking compared with cells not expressing this marker.The VGCC antagonist diltiazem and agonist (-)BayK 8644 were used to manipulate calcium activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, College of William and Mary, Williamsburg, Virginia, 23185.

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Sample image set from calcium imaging. A: Average brightfield image of cells during calcium imaging. B: Average Fluo4 fluorescence image of cells during calcium imaging. Bright regions indicate cells with high levels of calcium transients. C: Fluorescent image of cells after FISH. Positive cells are circled. D: Sample calcium activity data from a region of interest (ROI).
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fig06: Sample image set from calcium imaging. A: Average brightfield image of cells during calcium imaging. B: Average Fluo4 fluorescence image of cells during calcium imaging. Bright regions indicate cells with high levels of calcium transients. C: Fluorescent image of cells after FISH. Positive cells are circled. D: Sample calcium activity data from a region of interest (ROI).

Mentions: As the only VGCC α1 subunits expressed in cultured cells dissected at neural plate, neural fold, and neural tube stages, Cav1.2, Cav1.3, Cav2.1, Cav2.2, and Cav3.2 were examined further in calcium activity imaging experiments (Fig. 6). Two-hour calcium images were performed on neuronal cell culture, and spiking data were analyzed in three 40-minute time blocks. The number of calcium spikes in cells with VGCC expression (positive cells) was compared with the number of spikes in cells without detectable expression (negative cells). Cav2.1 is correlated with high-frequency calcium activity in cell cultures dissected at the neural plate and neural fold stages (Fig. 7A) and Cav22.22 is correlated with high-frequency calcium activity in neural tube cultures (Fig. 7B). The Cav1.2-, Cav1.3-, and Cav3.2-positive cells have equal spiking activity compared with cells negative for these markers.


The role of voltage-gated calcium channels in neurotransmitter phenotype specification: Coexpression and functional analysis in Xenopus laevis.

Lewis BB, Miller LE, Herbst WA, Saha MS - J. Comp. Neurol. (2014)

Sample image set from calcium imaging. A: Average brightfield image of cells during calcium imaging. B: Average Fluo4 fluorescence image of cells during calcium imaging. Bright regions indicate cells with high levels of calcium transients. C: Fluorescent image of cells after FISH. Positive cells are circled. D: Sample calcium activity data from a region of interest (ROI).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4043876&req=5

fig06: Sample image set from calcium imaging. A: Average brightfield image of cells during calcium imaging. B: Average Fluo4 fluorescence image of cells during calcium imaging. Bright regions indicate cells with high levels of calcium transients. C: Fluorescent image of cells after FISH. Positive cells are circled. D: Sample calcium activity data from a region of interest (ROI).
Mentions: As the only VGCC α1 subunits expressed in cultured cells dissected at neural plate, neural fold, and neural tube stages, Cav1.2, Cav1.3, Cav2.1, Cav2.2, and Cav3.2 were examined further in calcium activity imaging experiments (Fig. 6). Two-hour calcium images were performed on neuronal cell culture, and spiking data were analyzed in three 40-minute time blocks. The number of calcium spikes in cells with VGCC expression (positive cells) was compared with the number of spikes in cells without detectable expression (negative cells). Cav2.1 is correlated with high-frequency calcium activity in cell cultures dissected at the neural plate and neural fold stages (Fig. 7A) and Cav22.22 is correlated with high-frequency calcium activity in neural tube cultures (Fig. 7B). The Cav1.2-, Cav1.3-, and Cav3.2-positive cells have equal spiking activity compared with cells negative for these markers.

Bottom Line: Calcium activity was also analyzed on a single-cell level, and spike frequency was correlated with the expression of VGCC α1 subunits in cell culture.Cells expressing Cav 2.1 and Cav 2.2 displayed increased calcium spiking compared with cells not expressing this marker.The VGCC antagonist diltiazem and agonist (-)BayK 8644 were used to manipulate calcium activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, College of William and Mary, Williamsburg, Virginia, 23185.

Show MeSH
Related in: MedlinePlus