Dataset Information

Ca²⁺ -regulated cell migration revealed by optogenetically engineered Ca²⁺ oscillations.

ABSTRACT: The ability of a single Ca²⁺ ion to play an important role in cell biology is highlighted by the need for cells to form Ca²⁺ signals in the dimensions of space, time, and amplitude. Thus, spatial and temporal changes in intracellular Ca²⁺ concentration are important for determining cell fate. Optogenetic technology has been developed to provide more precise and targeted stimulation of cells. Here, U2OS cells overexpressing Ca²⁺ translocating channelrhodopsin (CatCh) were used to mediate Ca²⁺ influx through blue light illumination with various parameters, such as intensity, frequency, duty cycle, and duration. We identified that several Ca²⁺ -dependent transcription factors and certain kinases can be activated by specific Ca²⁺ waves. Using a wound-healing assay, we found that low-frequency Ca²⁺ oscillations increased cell migration through the activation of NF-κB. This study explores the regulation of cell migration by Ca²⁺ signals. Thus, we can choose optical parameters to modulate Ca²⁺ waves and achieve activation of specific signaling pathways. This novel methodology can be applied to clarify related cell-signaling mechanisms in the future.

SUBMITTER: Lai YS

PROVIDER: S-EPMC8048425 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Ca2+ -regulated cell migration revealed by optogenetically engineered Ca2+ oscillations.

Lai Yi-Shyun YS Chang Ya-Han YH Chen Yong-Yi YY Xu Jixuan J Yu Chi-Sian CS Chang Su-Jing SJ Chen Pai-Sheng PS Tsai Shaw-Jenq SJ Chiu Wen-Tai WT

Journal of cellular physiology 20201126 6

The ability of a single Ca2+ ion to play an important role in cell biology is highlighted by the need for cells to form Ca2+ signals in the dimensions of space, time, and amplitude. Thus, spatial and temporal changes in intracellular Ca2+ concentration are important for determining cell fate. Optogenetic technology has been developed to provide more precise and targeted stimulation of cells. Here, U2OS cells overexpressing Ca2+ translocating channelrho ...[more]

PMID: 33244795

Similar Datasets

Project description:ObjectiveHepatic Ca2+ signaling has been identified as a crucial key factor in driving gluconeogenesis. The involvement of mitochondria in hormone-induced Ca2+ signaling and their contribution to metabolic activity remain, however, poorly understood. Moreover, the molecular mechanism governing the mitochondrial Ca2+ efflux signaling remains unresolved. This study investigates the role of the Na+/Ca2+ exchanger, NCLX, in modulating hepatic mitochondrial Ca2+ efflux, and examines its physiological significance in hormonal hepatic Ca2+ signaling, gluconeogenesis, and mitochondrial bioenergetics.MethodsPrimary mouse hepatocytes from both an AAV-mediated conditional hepatic-specific and a total mitochondrial Na+/Ca2+ exchanger, NCLX, knockout (KO) mouse models were employed for fluorescent monitoring of purinergic and glucagon/vasopressin-dependent mitochondrial and cytosolic hepatic Ca2+ responses in cultured hepatocytes. Isolated liver mitochondria and permeabilized primary hepatocytes were used to analyze the ion-dependence of Ca2+ efflux. Utilizing the conditional hepatic-specific NCLX KO model, the rate of gluconeogenesis was assessed by first monitoring glucose levels in fasted mice, and subsequently subjecting the mice to a pyruvate tolerance test while monitoring their blood glucose. Additionally, cultured primary hepatocytes from both genotypes were assessed in vitro for glucagon-dependent glucose production and cellular bioenergetics through glucose oxidase assay and Seahorse respirometry, respectively.ResultsAnalysis of Ca2+ responses in isolated liver mitochondria and cultured primary hepatocytes from NCLX KO versus WT mice showed that NCLX serves as the principal mechanism for mitochondrial calcium extrusion in hepatocytes. We then determined the role of NCLX in glucagon and vasopressin-induced Ca2+ oscillations. Consistent with previous studies, glucagon and vasopressin triggered Ca2+ oscillations in WT hepatocytes, however, the deletion of NCLX resulted in selective elimination of mitochondrial, but not cytosolic, Ca2+ oscillations, underscoring NCLX's pivotal role in mitochondrial Ca2+ regulation. Subsequent in vivo investigation for hepatic NCLX role in gluconeogenesis revealed that, as opposed to WT mice which maintained normoglycemic blood glucose levels when fasted, conditional hepatic-specific NCLX KO mice exhibited a faster drop in glucose levels, becoming hypoglycemic. Furthermore, KO mice showed deficient conversion of pyruvate to glucose when challenged under fasting conditions. Concurrent in vitro assessments showed impaired glucagon-dependent glucose production and compromised bioenergetics in KO hepatocytes, thereby underscoring NCLX's significant contribution to hepatic glucose metabolism.ConclusionsThe study findings demonstrate that NCLX acts as the primary Ca2+ efflux mechanism in hepatocytes. NCLX is indispensable for regulating hormone-induced mitochondrial Ca2+ oscillations, mitochondrial metabolism, and sustenance of hepatic gluconeogenesis.

Dataset Information

Ca²⁺ -regulated cell migration revealed by optogenetically engineered Ca²⁺ oscillations.

Publications

Ca<sup>2+</sup> -regulated cell migration revealed by optogenetically engineered Ca<sup>2+</sup> oscillations.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets