Location, dynamics and solvent relaxation of a nile red-based phase-sensitive fluorescent membrane probe

Saxena, Roopali; Shrivastava, Sandeep; Haldar, Sourav; Klymchenko, Andrey S.; Chattopadhyay, Amitabha

doi:10.1016/j.chemphyslip.2014.04.007

Cited by 29 publications

(34 citation statements)

References 51 publications

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“…The advantage of NR12S over Nile Red is that it has unique orientation and location in the membrane due to the additional fatty acyl chain that helps in anchoring of the molecule in the membrane [19]. We recently showed, using the parallax approach [34], that the fluorophore in NR12S is localized at the membrane interface, characterized by an average depth of penetration of~18 Å from the center of the bilayer [16]. In addition, NR12S has the advantage of being localized exclusively in the outer leaflet of the membrane [35,36].…”

Section: Resultsmentioning

confidence: 97%

“…In addition, NR12S has the advantage of being localized exclusively in the outer leaflet of the membrane [35,36]. More importantly, NR12S fluorescence displays sensitivity to the membrane phase [16,19].…”

Section: Resultsmentioning

confidence: 98%

“…The fluorescence emission maximum was observed at~581 and 591 nm in native and cholesterol-depleted hippocampal membranes, respectively. The red shift in the emission spectrum of NR12S could be due to reduction in liquid-ordered phase induced by cholesterol depletion [16,19].…”

Section: Resultsmentioning

confidence: 99%

“…We recently reported the location, dynamics, and environmentsensitive properties of a novel Nile Red-based phase-sensitive membrane probe (NR12S) [16]. We further showed, utilizing model membranes of varying phases, that important fluorescence parameters such as emission maximum, red edge excitation shift (REES), anisotropy and lifetime of NR12S exhibit sensitivity to the membrane phase.…”

Section: Introductionmentioning

confidence: 98%

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Cholesterol-induced changes in hippocampal membranes utilizing a phase-sensitive fluorescence probe

Saxena

Shrivastava

Chattopadhyay

2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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The function of membrane receptors in the nervous system depends on physicochemical characteristics of neuronal membranes such as membrane order and phase. In this work, we have monitored the changes in hippocampal membrane order and related parameters by cholesterol and protein content utilizing a Nile Red-based phase-sensitive fluorescent membrane probe NR12S. Since alteration of membrane cholesterol is often associated with membrane phase change, the phase-sensitive nature of NR12S fluorescence becomes useful in these experiments. Our results show that fluorescence spectroscopic parameters such as emission maximum, anisotropy, and lifetime of NR12S display characteristic dependence on membrane cholesterol content. Interestingly, cholesterol-dependent red edge excitation shift is displayed by NR12S under these conditions. Hippocampal membranes exhibited reduction in liquid-ordered phase upon cholesterol depletion. These results provide insight into changes in hippocampal membrane order in the overall context of cholesterol and protein modulation.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

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Cholesterol-induced changes in hippocampal membranes utilizing a phase-sensitive fluorescence probe

Saxena

Shrivastava

Chattopadhyay

2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…Knowledge of the penetration depth of the fluorescent moiety in a membrane probe is important because it allows to correlate the fluorescence parameters of the probe with a defined location in the membrane (Haldar et al, 2012;Saxena et al, 2014). The penetration depth of the BODIPY fluorophore of BODIPY-C12 in DOPC membranes was determined by the parallax method (Chattopadhyay and London, 1987) using the equation:…”

Section: Resultsmentioning

confidence: 99%

Molecular rheology of neuronal membranes explored using a molecular rotor: Implications for receptor function

Pal

Chakraborty

Bandari

et al. 2016

Chemistry and Physics of Lipids

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The role of membrane cholesterol as a crucial regulator in the structure and function of membrane proteins and receptors is well documented. However, there is a lack of consensus on the mechanism for such regulation. We have previously shown that the function of an important neuronal receptor, the serotonin1A receptor, is modulated by cholesterol in hippocampal membranes. With an overall objective of addressing the role of 3 membrane physical properties in receptor function, we measured the viscosity of hippocampal membranes of varying cholesterol content using a meso-substituted fluorophore (BODIPY-C12) based on the BODIPY probe. BODIPY-C12 acts as a fluorescent molecular rotor and allows measurement of hippocampal membrane viscosity through its characteristic viscosity-sensitive fluorescence depolarization. A striking feature of our results is that specific agonist binding by the serotonin1A receptor exhibits close correlation with hippocampal membrane viscosity, implying the importance of global membrane properties in receptor function. We envision that our results are important in understanding GPCR regulation by the membrane environment, and is relevant in the context of diseases in which GPCR signaling plays a major role and are characterized by altered membrane fluidity.Abbreviations: 2-AS, 2-(9-anthroyloxy)stearic acid; 12-AS, 12-(9-anthroyloxy)stearic acid; BODIPY, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; GPCR, G protein-coupled receptor; 8-12-PC, 1-palmitoyl-2-(12-doxyl)stearoyl-sn-glycero-3-phosphocholine; DMPC, 1,2-dimyristoyl-sn-glycero-3-phosphocholine; DOPC, 1,2-dioleoylsn-glycero-3-phosphocholine; MβCD, methyl-β-cyclodextrin; Tempo-PC, 1,2-dioleoyl-snglycero-3-phosphotempocholine Keywords: Molecular rotor; hippocampal membrane; viscosity; cholesterol; neuronal receptor IntroductionBiological membranes are complex, highly organized, two-dimensional, supramolecular assemblies of a diverse variety of lipids and proteins. The function of membranes is to allow cellular compartmentalization, and impart an identity to individual cells and organelles, besides providing an appropriate environment for proper functioning of membrane proteins. Interestingly, cellular membranes in the nervous system are characterized by very high concentration and remarkable diversity of lipids, and these are correlated with increased complexity in the function of the nervous system (Sastry, 1985;Wenk, 2005). In this context, cholesterol represents an important lipid since brain cholesterol has been implicated in a number of neurological disorders (Chattopadhyay and Paila, 2007;Martín et al., 2014), some of which share a common etiology of defective cholesterol 4 metabolism in the brain (Porter and Herman, 2011). More importantly, the function of neuronal receptors depends on cholesterol (Pucadyil and Chattopadhyay, 2006;Allen et al., 2007;Paila and Chattopadhyay, 2010;Jafurulla and Chattopadhyay, 2013), which affects neurotransmission, resulting in mood and anxiety disorders (Papakostas et al., 2004). In spite of ...

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A Chemogenetic Approach for the Optical Monitoring of Voltage in Neurons

et al. 2019

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Optical monitoring of neuronal voltage using fluorescent indicators is ap owerfula pproach for the interrogation of the cellular and molecular logic of the nervous system. Herein, as emisynthetic tethered voltage indicator (STeVI1) based upon nile red is described that displays voltage sensitivity when genetically targeted to neuronal membranes. This environmentally sensitive probe allows for wash-free imaging and faithfully detects supra-and sub-threshold activity in neurons.

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Location, dynamics and solvent relaxation of a nile red-based phase-sensitive fluorescent membrane probe

Cited by 29 publications

References 51 publications

Cholesterol-induced changes in hippocampal membranes utilizing a phase-sensitive fluorescence probe

Cholesterol-induced changes in hippocampal membranes utilizing a phase-sensitive fluorescence probe

Molecular rheology of neuronal membranes explored using a molecular rotor: Implications for receptor function

A Chemogenetic Approach for the Optical Monitoring of Voltage in Neurons

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