Journal: Nature Communications

Article Title: Ca V 1.1 voltage-sensing domain III exclusively controls skeletal muscle excitation-contraction coupling

doi: 10.1038/s41467-024-51809-5

Figure Lengend Snippet: A Protein structure (top view) showing the four-fold symmetry of Ca V 1.1 with four voltage-sensing domains (VSD) arranged around the common channel pore (PD). B The Ca V 1.1 membrane topology reveals four homologous repeats, each with six membrane helices (S1–S6) (colors as in A ). C A VSD contains four transmembrane helices (S1–S4). S4 carries the regularly spaced positive gating charges (blue bands), which upon state transition successively interact with negative countercharges in the S2 and S3 helices (red bands) of the intracellular and extracellular negative clusters (INC, ENC). HCS indicates the hydrophobic constriction site. D The number and type of gating charges (R or K) differ between the S4 helices of the four VSDs. E In skeletal muscle Ca V 1.1 functions as calcium channel as well as voltage sensor for EC-coupling. F Here we simultaneously measure calcium currents and fluorescence calcium signals (EC-coupling) in dysgenic (Ca V 1.1-null) myotubes, reconstituted with different Ca V 1.1 constructs.

Article Snippet: Simultaneously to electrical stimulation, the myoplasmic calcium concentration was recorded in reconstituted GLT myotubes utilizing a fluorescence calcium indicator (Calbryte 590 potassium salt; AAT Bioquest) included to the intracellular solution, at a concentration of 0.2 mM.

Techniques: Membrane, Fluorescence, Construct

Journal: MethodsX

Article Title: Bayesian inference of molecular kinetic parameters from astrocyte calcium imaging data

doi: 10.1016/j.mex.2022.101825

Figure Lengend Snippet:

Article Snippet: Two weeks later, the cells were transduced with adeno-associated viral vectors (AAV; multiplicity of infection of 100,000) expressing the fluorescent intracellular calcium indicator fusion protein, GCaMP6m (SignaGen; Frederick, MD) in accordance with the supplier's protocol.

Techniques: Imaging, Plasmid Preparation, Microscopy, Software, Isolation, Transduction, Activity Assay, Modification