The Experts below are selected from a list of 174 Experts worldwide ranked by ideXlab platform
Richard E Mccarty - One of the best experts on this subject based on the ideXlab platform.
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influences of energization and nucleotide binding on the reaction of Lucifer Yellow vinyl sulfone with the alpha subunits of the chloroplast atp synthase
Biochemistry, 2000Co-Authors: Kristina Cunningham, Richard E MccartyAbstract:The catalytic portion of the chloroplast ATP synthase (CF1) consists of five different polypeptides in the stoichiometry α3β3γδe and is structurally asymmetric. Asymmetry is readily apparent in the properties of the six nucleotide binding sites and the single-copy, smaller subunits. Asymmetry is also detected in the α subunits by the rapid and covalent binding of Lucifer Yellow vinyl sulfone (LY) to one of the three chemically identical α subunits. The binding of LY to a single α subunit has allowed the investigation of whether asymmetry in the α subunits is a permanent feature of CF1. The development of an electrochemical proton gradient across illuminated thylakoid membranes and the preincubation of CF1 in solution with Mg2+-ATP were found to alter the LY distribution such that multiple α subunits were labeled with LY. Illumination of thylakoid membranes doubled the extent of LY labeling, and fluorescence resonance energy transfer measurements indicated that LY was bound to more than one α subunit. Sinc...
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asymmetry of the alpha subunit of the chloroplast atp synthase as probed by the binding of Lucifer Yellow vinyl sulfone
Biochemistry, 1998Co-Authors: Kristina M Lowe, Richard E MccartyAbstract:The catalytic portion of the chloroplast ATP synthase (CF1) is structurally asymmetric. Asymmetry of the otherwise symmetrical alpha3beta3 heterohexamer is induced by the presence of tightly bound nucleotides and interactions with the single-copy, smaller subunits. Lucifer Yellow vinyl sulfone (4-amino-N-[3-(vinylsulfonyl)phenyl]naphthalimide-3,6-disulfonic acid) rapidly and covalently binds to lysine 378 on one alpha subunit [Nalin, C. M., Snyder, B., and McCarty, R. E., (1985) Biochemistry 24, 2318-2324] [Shapiro, A. B. (1991) Ph.D. Thesis, Cornell University, Ithaca, NY). The asymmetrical binding of Lucifer Yellow to CF1 provides a method to investigate the cause of asymmetry in the alpha subunits. The reaction of CF1 with Lucifer Yellow was monitored by total fluorescence of bound Lucifer Yellow as well as by quantitative determination of Lucifer Yellow bound to the tryptic peptide that contains lysine 378 of the alpha subunit. The total binding of Lucifer Yellow to CF1 was not affected by the presence of tightly bound nucleotides or nucleotide in the medium. Neither the total binding of Lucifer Yellow to CF1 nor the reaction of alpha-lysine 378 with Lucifer Yellow was changed by the removal of the epsilon subunit, the delta subunit, or both subunits. The extent of incorporation of Lucifer Yellow into lysine 378 of the alpha subunit in (alphabeta)n was about three times that of Lucifer Yellow incorporation into CF1. Reconstitution of (alphabeta)n with gamma restored the binding of one Lucifer Yellow per alpha3beta3gamma. Therefore, the interactions between gamma and the alphabeta heterohexamer are important in conferring asymmetry to the alpha subunits of CF1.
Annica Dahlstrom - One of the best experts on this subject based on the ideXlab platform.
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studies on the 3 dimensional architecture of dendritic spines and varicosities in human cortex by confocal laser scanning microscopy and Lucifer Yellow microinjections
Journal of Neuroscience Methods, 1995Co-Authors: Pavel V Belichenko, Annica DahlstromAbstract:Abstract A method for 3-dimensional (3-D) visualization of dendritic spines and varicosities in human cortical neurons is described. Intracellular microinjection of Lucifer Yellow was used to display the morphology of dendrites on pyramidal and non-pyramidal neurons. Confocal laser scanning microscopy was used for imaging, and 3-D reconstructions and analysis of spines and varicosities were performed. The frontal, temporal, parietal and occipital cortices, and hippocampus in normal and pathological human brains were studied. Using this technique spines can be visualized from both sides of dendrites, which are ‘hidden’ in 2-D representations, and therefore not usually included in the extimation of dendritic spine density/total spine numbers. In patients with Rett's syndrome and some epilepsy patients, a regional loss of dendritic spines (‘naked’ dendrites) was found. These results will be included in the Human Brain Mapping Project.
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dual channel confocal laser scanning microscopy of Lucifer Yellow microinjected human brain cells combined with texas red immunofluorescence
Journal of Neuroscience Methods, 1994Co-Authors: Pavel V Belichenko, Annica DahlstromAbstract:A method for visualization of individual human brain cells and their dendritic extensions in combination with immunofluorescence is described. Microinjection of Lucifer Yellow was used to reveal the dendritic morphology of cortical brain cells. Indirect immunofluorescence with Texas Red as label was used to investigate the distribution of 3 different groups of immunogens: enzymes (monoamine oxidase A and B), receptors (beta-adrenoceptor protein), and synaptic vesicle proteins (synapsin I and synaptophysin) in each cortical slice. A dual-channel confocal laser scanning microscope with an argon/krypton laser was used for imaging these double-stained fluorescent specimens. Lucifer Yellow and Texas Red were recorded simultaneously or separately, taking advantage of the different activating lines (488 lambda and 568 lambda) of the laser and using the two filter blocks (K1 and K2) supplied with the instrument (BioRad MRC-600) for recording the emission of either fluorophore. Using this technique we have demonstrated the localization of immunoreactive material in relation to the dendritic morphology of cortical cells.
K Negishi - One of the best experts on this subject based on the ideXlab platform.
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double staining of horizontal and amacrine cells by intracellular injection with Lucifer Yellow and biocytin in carp retina
Neuroscience, 1994Co-Authors: T Teranishi, K NegishiAbstract:Abstract Horizontal and amacrine cells in the isolated carp retina were impaled with micropipette electrode, identified by their characteristic light responses, and injected iontophoretically with markers for morphological study. Both Lucifer Yellow CH and biocytin were injected simultaneously. Lucifer Yellow was seen by its own fluorescence while biocytin was visualized by binding with Texas Red-linked or horseradish peroxidase-conjugated avidin. For cone-connected horizontal cells, biocytin-coupled cells were found to be approximately five-times more numerous than Lucifer Yellow-coupled cells. Coupling for both tracers was consistently hampered by intravitreally applied dopamine. In untreated retinas, the injected Lucifer Yellow was restricted within one rod-connected horizontal cell, while biocytin revealed several coupled neighbors. Amacrine cells, labeled by the tracers, were morphologically grouped into eight types, based on our earlier classification.29 Among them, amacrine cells, belonging to three types (Fnd, Pmb or Pma), were confirmed to be Lucifer Yellow-coupled, and the number of biocytin-coupled cells was more numreous (about 2.5 times) than that of Lucifer Yellow-coupled cells. Most amacrine cells (i.e. Pwd, Fnb and Fna) showed biocytin-coupling with no Lucifer Yellow-coupling. A few classified (i.e. Pwb and Fwa) and unclassified cells did not show any coupling. Since the tracer coupling takes place via gap junctions, the majority of amacrine cells, belonging to certain homologous types, appear to be functionally coupled with each other in the inner plexiform layer. However, dopamine did not influence the range of tracer coupling between amacrine cells in the carp retina under the present experimental conditions.
Pavel V Belichenko - One of the best experts on this subject based on the ideXlab platform.
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studies on the 3 dimensional architecture of dendritic spines and varicosities in human cortex by confocal laser scanning microscopy and Lucifer Yellow microinjections
Journal of Neuroscience Methods, 1995Co-Authors: Pavel V Belichenko, Annica DahlstromAbstract:Abstract A method for 3-dimensional (3-D) visualization of dendritic spines and varicosities in human cortical neurons is described. Intracellular microinjection of Lucifer Yellow was used to display the morphology of dendrites on pyramidal and non-pyramidal neurons. Confocal laser scanning microscopy was used for imaging, and 3-D reconstructions and analysis of spines and varicosities were performed. The frontal, temporal, parietal and occipital cortices, and hippocampus in normal and pathological human brains were studied. Using this technique spines can be visualized from both sides of dendrites, which are ‘hidden’ in 2-D representations, and therefore not usually included in the extimation of dendritic spine density/total spine numbers. In patients with Rett's syndrome and some epilepsy patients, a regional loss of dendritic spines (‘naked’ dendrites) was found. These results will be included in the Human Brain Mapping Project.
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dual channel confocal laser scanning microscopy of Lucifer Yellow microinjected human brain cells combined with texas red immunofluorescence
Journal of Neuroscience Methods, 1994Co-Authors: Pavel V Belichenko, Annica DahlstromAbstract:A method for visualization of individual human brain cells and their dendritic extensions in combination with immunofluorescence is described. Microinjection of Lucifer Yellow was used to reveal the dendritic morphology of cortical brain cells. Indirect immunofluorescence with Texas Red as label was used to investigate the distribution of 3 different groups of immunogens: enzymes (monoamine oxidase A and B), receptors (beta-adrenoceptor protein), and synaptic vesicle proteins (synapsin I and synaptophysin) in each cortical slice. A dual-channel confocal laser scanning microscope with an argon/krypton laser was used for imaging these double-stained fluorescent specimens. Lucifer Yellow and Texas Red were recorded simultaneously or separately, taking advantage of the different activating lines (488 lambda and 568 lambda) of the laser and using the two filter blocks (K1 and K2) supplied with the instrument (BioRad MRC-600) for recording the emission of either fluorophore. Using this technique we have demonstrated the localization of immunoreactive material in relation to the dendritic morphology of cortical cells.
Tomas Ho Ffelt - One of the best experts on this subject based on the ideXlab platform.
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putative gabaergic input to axons of spinal interneurons and primary sensory neurons in the lamprey spinal cord as shown by intracellular Lucifer Yellow and gaba immunohistochemistry
Brain Research, 1991Co-Authors: Johan Christenson, Sten Grillner, Fulvia Bongianni, Tomas Ho FfeltAbstract:Abstract GABAergic plastic modulation of the membrane potential occurs in spinal interneurons during fictive locomotion in lamprey presumably indicating a presynaptic inhibition. GABA also modulates synaptic transmission from primary sensory neurons (dorsal cells) at a presynaptic site. From these findings GABA terminals would be expected to be in close contact with phasically modulated axons of spinal interneurons and/or dorsal cells and their axons. To test this supposition intracellular injections of Lucifer Yellow into spinal interneurons or dorsal cells were combined with GABA immunohistochemistry. GABA-immunoreactive (ir) varicosities were found to be in close contact (
