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In vitro and in vivo calibration of low affinity genetic Ca 2+ indicators
We report here a procedure to calibrate the ratiometric signal of GAP (GFP-Aequorin Protein) targeted to the endo-sarcoplasmic reticulum (ER/SR) into [Ca2+]ER/SR based on imaging of fluorescence after heating the tissue at 50-52 °C, since this value coincides with that obtained in the absence of Ca2+ (Rmin). Knowledge of the dynamic range (Rmax/Rmin) and the Ca2+-affinity (KD) of the indicator permits calculation of [Ca2+] by applying a simple algorithm. We have validated this procedure in vitro using several cell types (HeLa, HEK 293T and mouse astrocytes), as well as in vivo in Drosophila. Moreover, this methodology is ...
Source: Cell Calcium - November 13, 2023 Category: Cytology Authors: Alba Delrio-Lorenzo Jonathan Rojo-Ruiz Patricia Torres-Vidal Maria Teresa Alonso Javier Garc ía-Sancho Source Type: research

Subcellular localization and transcriptional regulation of brain ryanodine receptors. Functional implications
Cell Calcium. 2023 Nov 4;116:102821. doi: 10.1016/j.ceca.2023.102821. Online ahead of print.ABSTRACTRyanodine receptors (RyR) are intracellular Ca2+ channels localized in the endoplasmic reticulum, where they act as critical mediators of Ca2+-induced Ca2+ calcium release (CICR). In the brain, mammals express in both neurons, and non-neuronal cells, a combination of the three RyR-isoforms (RyR1-3). Pharmacological approaches, which do not distinguish between isoforms, have indicated that RyR-isoforms contribute to brain function. However, isoform-specific manipulations have revealed that RyR-isoforms display different subce...
Source: Cell Calcium - November 10, 2023 Category: Cytology Authors: Rodrigo Torres Cecilia Hidalgo Source Type: research

Neuronal and astrocyte NCX isoform/splice variants: How do they participate in Na + and Ca 2+ signalling?
Cell Calcium. 2023 Oct 24;116:102818. doi: 10.1016/j.ceca.2023.102818. Online ahead of print.ABSTRACTNCX1, NCX2, and NCX3 gene isoforms and their splice variants are characteristically expressed in different regions of the brain. The tissue-specific splice variants of NCX1-3 isoforms show specific expression profiles in neurons and astrocytes, whereas the relevant NCX isoform/splice variants exhibit diverse allosteric modes of Na+- and Ca2+-dependent regulation. In general, overexpression of NCX1-3 genes leads to neuroprotective effects, whereas their ablation gains the opposite results. At this end, the partial contributi...
Source: Cell Calcium - November 2, 2023 Category: Cytology Authors: Daniel Khananshvili Source Type: research

Calcium transport and sensing in TRPC channels - New insights into a complex feedback regulation
Cell Calcium. 2023 Oct 18;116:102816. doi: 10.1016/j.ceca.2023.102816. Online ahead of print.ABSTRACTCanonical TRP (TRPC) channels are a still enigmatic family of signaling molecules with multimodal sensing features. These channels enable Ca2+ influx through the plasma membrane to control a diverse range of cellular functions. Based on both regulatory- and recently uncovered structural features, TRPC channels are considered to coordinate Ca2+ and other divalent cations not only within the permeation path but also at additional sensory sites. Analysis of TRPC structures by cryo-EM identified multiple regulatory ion binding ...
Source: Cell Calcium - October 28, 2023 Category: Cytology Authors: Jasmin Baron Klaus Groschner Oleksandra Tiapko Source Type: research