TRH-receptor mobility and function in intact and cholesterol-depleted plasma membrane of HEK293 cells stably expressing TRH-R-eGFP

Brejchova, Jana; Sýkora, Jan; Ostasov, Pavel; Merta, Ladislav; Roubalová, Lenka; Janáček, Jiřı́; Hof, Martin

doi:10.1016/j.bbamem.2014.11.029

Cited by 19 publications

(18 citation statements)

References 84 publications

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“…Removal of cholesterol by β-MCD reduced lateral mobility of FP receptors and increased residence time. These data are in agreement with previous studies where acute depletion of cholesterol reduces mobility of membrane proteins and increases confinement [53,54]. …”

Section: Discussionsupporting

confidence: 93%

“…The concentration of β-MCD used in this study removes approximately 80% of the cholesterol [55] which would allow for packing of acyl chains reducing fluidity of the membrane and most likely lateral mobility of the FP receptor. Acute depletion of cholesterol in the present study also increased residence time of FP receptors which has been shown in previous studies in which acute or chronic depletion of cholesterol increased confinement of membrane-bound proteins [53,54]. Depletion of membrane cholesterol results in reorganization of the cytoskeleton and phosphatidylinositol which may lead to increased confinement of membrane-bound receptors.…”

Section: Discussionsupporting

confidence: 86%

See 1 more Smart Citation

Effect of fish oil on lateral mobility of prostaglandin F2α (FP) receptors and spatial distribution of lipid microdomains in bovine luteal cell plasma membrane in vitro

Plewes

Burns

Graham

et al. 2017

Domestic Animal Endocrinology

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Lipid microdomains are ordered regions on the plasma membrane of cells, rich in cholesterol and sphingolipids, ranging in size from 10 to 200 nm in diameter. These lipid-ordered domains may serve as platforms to facilitate colocalization of intracellular signaling proteins during agonist-induced signal transduction. It is hypothesized that fish oil will disrupt the lipid microdomains, increasing spatial distribution of these lipid-ordered domains and lateral mobility of the prostaglandin (PG) F2α (FP) receptors in bovine luteal cells. The objectives of this study were to examine the effects of fish oil on (1) the spatial distribution of lipid microdomains, (2) lateral mobility of FP receptors, and (3) lateral mobility of FP receptors in the presence of PGF2α on the plasma membrane of bovine luteal cells in vitro. Bovine ovaries were obtained from a local abattoir and corpora lutea were digested using collagenase. In experiment 1, lipid microdomains were labeled using cholera toxin subunit B Alexa Fluor 555. Domains were detected as distinct patches on the plasma membrane of mixed luteal cells. Fish oil treatment decreased fluorescent intensity in a dose-dependent manner (P < 0.01). In experiment 2, single particle tracking was used to examine the effects of fish oil treatment on lateral mobility of FP receptors. Fish oil treatment increased microdiffusion and macrodiffusion coefficients of FP receptors as compared to control cells (P < 0.05). In addition, compartment diameters of domains were larger, and residence times were reduced for receptors in fish oil–treated cells (P < 0.05). In experiment 3, single particle tracking was used to determine the effects of PGF2α on lateral mobility of FP receptors and influence of fish oil treatment. Lateral mobility of receptors was decreased within 5 min following the addition of ligand for control cells (P < 0.05). However, lateral mobility of receptors was unaffected by addition of ligand for fish oil–treated cells (P > 0.10). The data presented provide strong evidence that fish oil causes a disruption in lipid microdomains and affects lateral mobility of FP receptors in the absence and presence of PGF2α.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 86%

Effect of fish oil on lateral mobility of prostaglandin F2α (FP) receptors and spatial distribution of lipid microdomains in bovine luteal cell plasma membrane in vitro

Plewes

Burns

Graham

et al. 2017

Domestic Animal Endocrinology

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“…We obtained values for CD95 diffusion coefficient that were comparable with values found in the literature for other membrane receptors [62][63][64][65][66]. The values for the diffusion coefficient of CD95 obtained by FCS and FRAP were within the same order of magnitude and the results obtained with both techniques showed the same tendency; namely a decrease in CD95 diffusion when the cells were in contact with the agonistic antibody.…”

Section: Discussionsupporting

confidence: 87%

Early activation of CD95 is limited and localized to the cytotoxic synapse

et al. 2018

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The cytotoxic synapse formed between cytotoxic T lymphocytes or natural killer cells expressing CD95L and target cells with CD95 on their surface is a key pathway for apoptosis induction by the immune system. Despite similarities with the immune synapse in antigen presenting cells, little is known about the role of the spatiotemporal organization of agonistic proteins/receptor interactions for CD95 signaling. Here, we have developed an artificial cytotoxic synapse to examine how mobility and geometry of an anti-CD95 agonistic antibody affect receptor aggregation and mobility, ie the first step of receptor activation. By measuring the distribution, diffusion coefficient, and fraction of immobile CD95 receptor in living cells, we show that at short times, the initial activation of CD95 occurs locally and is limited to the contact region of the cytotoxic synapse. This anisotropic activation of apoptotic signaling supports a role for confined interactions on the efficiency of signal transduction that may have implications for biomedical applications of extrinsic apoptosis induction.

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“…RICS has been applied to analyze membrane proteins [ 25 , 26 ], DNA [ 27 ], and other biomolecules [ 28 , 29 ] because it provides the spatial distribution of the diffusional coefficients in the diffusion map [ 27 , 30 ]. Each brick constituting the diffusion map showed the diffusion property of biomolecules contained in each microdomain.…”

Section: Introductionmentioning

confidence: 99%

Negative Correlation between the Diffusion Coefficient and Transcriptional Activity of the Glucocorticoid Receptor

Mikuni

Yamamoto

Horio

et al. 2017

IJMS

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The glucocorticoid receptor (GR) is a transcription factor, which interacts with DNA and other cofactors to regulate gene transcription. Binding to other partners in the cell nucleus alters the diffusion properties of GR. Raster image correlation spectroscopy (RICS) was applied to quantitatively characterize the diffusion properties of EGFP labeled human GR (EGFP-hGR) and its mutants in the cell nucleus. RICS is an image correlation technique that evaluates the spatial distribution of the diffusion coefficient as a diffusion map. Interestingly, we observed that the averaged diffusion coefficient of EGFP-hGR strongly and negatively correlated with its transcriptional activities in comparison to that of EGFP-hGR wild type and mutants with various transcriptional activities. This result suggests that the decreasing of the diffusion coefficient of hGR was reflected in the high-affinity binding to DNA. Moreover, the hyper-phosphorylation of hGR can enhance the transcriptional activity by reduction of the interaction between the hGR and the nuclear corepressors.

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TRH-receptor mobility and function in intact and cholesterol-depleted plasma membrane of HEK293 cells stably expressing TRH-R-eGFP

Cited by 19 publications

References 84 publications

Effect of fish oil on lateral mobility of prostaglandin F2α (FP) receptors and spatial distribution of lipid microdomains in bovine luteal cell plasma membrane in vitro

Effect of fish oil on lateral mobility of prostaglandin F2α (FP) receptors and spatial distribution of lipid microdomains in bovine luteal cell plasma membrane in vitro

Early activation of CD95 is limited and localized to the cytotoxic synapse

Negative Correlation between the Diffusion Coefficient and Transcriptional Activity of the Glucocorticoid Receptor

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