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Article: Impact of administration route on nanocarrier biodistribution in a murine colitis model.

Applegate, Catherine C / Deng, Hongping / Kleszynski, Brittany L / Cross, Tzu-Wen L / Konopka, Christian J / Dobrucki, L Wawrzyniec / Nelson, Erik R / Wallig, Matthew A / Smith, Andrew M / Swanson, Kelly S

Journal of experimental nanoscience

2022 Volume 17, Issue 1, Page(s) 599–616

Abstract: The incidence of inflammatory bowel disease (IBD) is increasing worldwide. Although current diagnostic and disease monitoring tests for IBD sensitively detect gut inflammation, they lack the molecular and cellular specificity of positron emission ... ...

Abstract	The incidence of inflammatory bowel disease (IBD) is increasing worldwide. Although current diagnostic and disease monitoring tests for IBD sensitively detect gut inflammation, they lack the molecular and cellular specificity of positron emission tomography (PET). In this proof-of-concept study, we use a radiolabeled macrophage-targeted nanocarrier probe (
Language	English
Publishing date	2022-10-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2233966-8
ISSN	1745-8099 ; 1745-8080
ISSN (online)	1745-8099
ISSN	1745-8080
DOI	10.1080/17458080.2022.2134563
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Article: Variable-angle epifluorescence microscopy: a new way to look at protein dynamics in the plant cell cortex.

Konopka, Catherine A / Bednarek, Sebastian Y

The Plant journal : for cell and molecular biology

2008 Volume 53, Issue 1, Page(s) 186–196

Abstract: Live-cell microscopy imaging of fluorescent-tagged fusion proteins is an essential tool for cell biologists. Total internal reflection fluorescence microscopy (TIRFM) has joined confocal microscopy as a complementary system for the imaging of cell ... ...

Abstract	Live-cell microscopy imaging of fluorescent-tagged fusion proteins is an essential tool for cell biologists. Total internal reflection fluorescence microscopy (TIRFM) has joined confocal microscopy as a complementary system for the imaging of cell surface protein dynamics in mammalian and yeast systems because of its high temporal and spatial resolution. Here we present an alternative to TIRFM, termed variable-angle epifluorescence microscopy (VAEM), for the visualization of protein dynamics at or near the plasma membrane of plant epidermal cells and root hairs in whole, intact seedlings that provides high-signal, low-background and near real-time imaging. VAEM uses highly oblique subcritical incident angles to decrease background fluorophore excitation. We discuss the utilities and advantages of VAEM for imaging of fluorescent fusion-tagged marker proteins in studying cortical cytoskeletal and membrane proteins. We believe that the application of VAEM will be an invaluable imaging tool for plant cell biologists.
MeSH term(s)	Cell Membrane/ultrastructure ; Cytoplasm ; Microscopy, Fluorescence/methods ; Plant Cells ; Plant Proteins/analysis ; Plant Proteins/chemistry
Chemical Substances	Plant Proteins
Language	English
Publishing date	2008-01
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1088037-9
ISSN	1365-313X ; 0960-7412
ISSN (online)	1365-313X
ISSN	0960-7412
DOI	10.1111/j.1365-313X.2007.03306.x
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Article ; Online: An essential role for MEF2C in the cortical response to loss of sleep in mice.

Bjorness, Theresa E / Kulkarni, Ashwinikumar / Rybalchenko, Volodymyr / Suzuki, Ayako / Bridges, Catherine / Harrington, Adam J / Cowan, Christopher W / Takahashi, Joseph S / Konopka, Genevieve / Greene, Robert W

eLife

2020 Volume 9

Abstract: Neuronal activity and gene expression in response to the loss of sleep can provide a window into the enigma of sleep function. Sleep loss is associated with brain differential gene expression, an increase in pyramidal cell mEPSC frequency and amplitude, ... ...

Abstract	Neuronal activity and gene expression in response to the loss of sleep can provide a window into the enigma of sleep function. Sleep loss is associated with brain differential gene expression, an increase in pyramidal cell mEPSC frequency and amplitude, and a characteristic rebound and resolution of slow wave sleep-slow wave activity (SWS-SWA). However, the molecular mechanism(s) mediating the sleep-loss response are not well understood. We show that sleep-loss regulates MEF2C phosphorylation, a key mechanism regulating MEF2C transcriptional activity, and that MEF2C function in postnatal excitatory forebrain neurons is required for the biological events in response to sleep loss in C57BL/6J mice. These include altered gene expression, the increase and recovery of synaptic strength, and the rebound and resolution of SWS-SWA, which implicate MEF2C as an essential regulator of sleep function.
MeSH term(s)	Animals ; Cerebral Cortex/physiology ; Gene Expression Regulation ; MEF2 Transcription Factors/genetics ; MEF2 Transcription Factors/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Sleep/physiology ; Sleep Deprivation ; Transcription, Genetic
Chemical Substances	MEF2 Transcription Factors ; Mef2c protein, mouse
Language	English
Publishing date	2020-08-27
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.58331
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Article: Comparison of the dynamics and functional redundancy of the Arabidopsis dynamin-related isoforms DRP1A and DRP1C during plant development.

Konopka, Catherine A / Bednarek, Sebastian Y

Plant physiology

2008 Volume 147, Issue 4, Page(s) 1590–1602

Abstract: Members of the Arabidopsis (Arabidopsis thaliana) DYNAMIN-RELATED PROTEIN1 (DRP1) family are required for cytokinesis and cell expansion. Two isoforms, DRP1A and DRP1C, are required for plasma membrane maintenance during stigmatic papillae expansion and ... ...

Abstract	Members of the Arabidopsis (Arabidopsis thaliana) DYNAMIN-RELATED PROTEIN1 (DRP1) family are required for cytokinesis and cell expansion. Two isoforms, DRP1A and DRP1C, are required for plasma membrane maintenance during stigmatic papillae expansion and pollen development, respectively. It is unknown whether the DRP1s function interchangeably or if they have distinct roles during cell division and expansion. DRP1C was previously shown to form dynamic foci in the cell cortex, which colocalize with part of the clathrin endocytic machinery in plants. DRP1A localizes to the plasma membrane, but its cortical organization and dynamics have not been determined. Using dual color labeling with live cell imaging techniques, we showed that DRP1A also forms discreet dynamic foci in the epidermal cell cortex. Although the foci overlap with those formed by DRP1C and clathrin light chain, there are clear differences in behavior and response to pharmacological inhibitors between DRP1A and DRP1C foci. Possible functional or regulatory differences between DRP1A and DRP1C were supported by the failure of DRP1C to functionally compensate for the absence of DRP1A. Our studies indicated that the DRP1 isoforms function or are regulated differently during cell expansion.
MeSH term(s)	Amino Acid Sequence ; Arabidopsis/genetics ; Arabidopsis/growth & development ; Arabidopsis/metabolism ; Arabidopsis Proteins/analysis ; Arabidopsis Proteins/genetics ; Arabidopsis Proteins/physiology ; Cell Membrane/chemistry ; Cell Membrane/drug effects ; Conserved Sequence ; Cytoskeleton/drug effects ; Dynamins/analysis ; Dynamins/chemistry ; Dynamins/genetics ; Dynamins/metabolism ; Dynamins/physiology ; Flowers/genetics ; Flowers/growth & development ; Flowers/metabolism ; Genetic Complementation Test ; Green Fluorescent Proteins/analysis ; Phylogeny ; Phytosterols/metabolism ; Protein Isoforms/analysis ; Protein Isoforms/genetics ; Protein Isoforms/physiology ; Seedlings/genetics ; Seedlings/growth & development ; Seedlings/metabolism ; Sequence Analysis, Protein
Chemical Substances	Arabidopsis Proteins ; Phytosterols ; Protein Isoforms ; Green Fluorescent Proteins (147336-22-9) ; ADL1A protein, Arabidopsis (EC 3.6.5.5) ; DRP1C protein, Arabidopsis (EC 3.6.5.5) ; Dynamins (EC 3.6.5.5)
Language	English
Publishing date	2008-03-14
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	208914-2
ISSN	1532-2548 ; 0032-0889
ISSN (online)	1532-2548
ISSN	0032-0889
DOI	10.1104/pp.108.116863
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Article ; Online: Nanocarriers targeting adipose macrophages increase glucocorticoid anti-inflammatory potency to ameliorate metabolic dysfunction.

Prabhu, Suma / Deng, Hongping / Cross, Tzu-Wen L / Shahoei, Sayyed Hamed / Konopka, Christian J / Gonzalez Medina, Natalia / Applegate, Catherine C / Wallig, Matthew A / Dobrucki, L Wawrzyniec / Nelson, Erik R / Smith, Andrew M / Swanson, Kelly S

Biomaterials science

2020 Volume 9, Issue 2, Page(s) 506–518

Abstract: Obesity is associated with systemic inflammation due to macrophage accumulation in adipose tissue (AT). AT macrophages are, therefore, a target for therapeutics to modulate inflammation and prevent comorbidities. Because inflammatory processes have ... ...

Abstract	Obesity is associated with systemic inflammation due to macrophage accumulation in adipose tissue (AT). AT macrophages are, therefore, a target for therapeutics to modulate inflammation and prevent comorbidities. Because inflammatory processes have pleiotropic effects throughout the body and are intertwined with metabolic axes, systemic anti-inflammatory therapies are often harmful. We report that targeting AT macrophages using dextran nanocarriers radically alters the pharmacology of anti-inflammatory glucocorticoids, uncoupling the metabolic axis in obese mice. Following a single treatment, expression of inflammatory mediators and markers of inflammatory macrophages decreased with a nearly 20-fold higher potency compared with free drug. As a result, long-term treatment resulted in potent fat mobilization, AT reduction, weight loss, improved glucose tolerance, and altered AT gene expression profiles that led to elevated liver stress. Two weeks after treatment ceased, gene expression of inflammatory mediators in AT remained lower than obese controls, while gene expression related to metabolic function improved. These data demonstrate that nanocarriers show potential for amelioration of obesity-related AT inflammation and metabolic dysfunction, highlighting an important opportunity for nanomedicine to impact chronic metabolic disorders with complex and poorly understood etiology.
MeSH term(s)	Adipose Tissue ; Animals ; Anti-Inflammatory Agents/pharmacology ; Glucocorticoids ; Inflammation/drug therapy ; Insulin Resistance ; Macrophages ; Mice ; Mice, Inbred C57BL
Chemical Substances	Anti-Inflammatory Agents ; Glucocorticoids
Language	English
Publishing date	2020-11-17
Publishing country	England
Document type	Journal Article
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/d0bm01142h
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Article: Nanocarriers targeting adipose macrophages increase glucocorticoid anti-inflammatory potency to ameliorate metabolic dysfunction

Prabhu, Suma / Deng, Hongping / Cross, Tzu-Wen L / Shahoei, Sayyed Hamed / Konopka, Christian J / Gonzalez Medina, Natalia / Applegate, Catherine C / Wallig, Matthew A / Dobrucki, L. Wawrzyniec / Nelson, Erik R / Smith, Andrew M / Swanson, Kelly S

Biomaterials science. 2021 Jan. 26, v. 9, no. 2

2021

Abstract	Obesity is associated with systemic inflammation due to macrophage accumulation in adipose tissue (AT). AT macrophages are, therefore, a target for therapeutics to modulate inflammation and prevent comorbidities. Because inflammatory processes have pleiotropic effects throughout the body and are intertwined with metabolic axes, systemic anti-inflammatory therapies are often harmful. We report that targeting AT macrophages using dextran nanocarriers radically alters the pharmacology of anti-inflammatory glucocorticoids, uncoupling the metabolic axis in obese mice. Following a single treatment, expression of inflammatory mediators and markers of inflammatory macrophages decreased with a nearly 20-fold higher potency compared with free drug. As a result, long-term treatment resulted in potent fat mobilization, AT reduction, weight loss, improved glucose tolerance, and altered AT gene expression profiles that led to elevated liver stress. Two weeks after treatment ceased, gene expression of inflammatory mediators in AT remained lower than obese controls, while gene expression related to metabolic function improved. These data demonstrate that nanocarriers show potential for amelioration of obesity-related AT inflammation and metabolic dysfunction, highlighting an important opportunity for nanomedicine to impact chronic metabolic disorders with complex and poorly understood etiology.
Keywords	adipose tissue ; biochemical pathways ; biocompatible materials ; dextran ; drugs ; etiology ; gene expression ; glucocorticoids ; glucose tolerance ; inflammation ; liver ; macrophages ; nanocarriers ; nanomedicine ; obesity ; pharmacology ; weight loss
Language	English
Dates of publication	2021-0126
Size	p. 506-518.
Publishing place	The Royal Society of Chemistry
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/d0bm01142h
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Article ; Online: MEF2C Hypofunction in Neuronal and Neuroimmune Populations Produces MEF2C Haploinsufficiency Syndrome-like Behaviors in Mice.

Harrington, Adam J / Bridges, Catherine M / Berto, Stefano / Blankenship, Kayla / Cho, Jennifer Y / Assali, Ahlem / Siemsen, Benjamin M / Moore, Hannah W / Tsvetkov, Evgeny / Thielking, Acadia / Konopka, Genevieve / Everman, David B / Scofield, Michael D / Skinner, Steven A / Cowan, Christopher W

Biological psychiatry

2020 Volume 88, Issue 6, Page(s) 488–499

Abstract: Background: Microdeletions of the MEF2C gene are linked to a syndromic form of autism termed MEF2C haploinsufficiency syndrome (MCHS). MEF2C hypofunction in neurons is presumed to underlie most of the symptoms of MCHS. However, it is unclear in which ... ...

Abstract	Background: Microdeletions of the MEF2C gene are linked to a syndromic form of autism termed MEF2C haploinsufficiency syndrome (MCHS). MEF2C hypofunction in neurons is presumed to underlie most of the symptoms of MCHS. However, it is unclear in which cell populations MEF2C functions to regulate neurotypical development. Methods: Multiple biochemical, molecular, electrophysiological, behavioral, and transgenic mouse approaches were used to characterize MCHS-relevant synaptic, behavioral, and gene expression changes in mouse models of MCHS. Results: We showed that MCHS-associated missense mutations cluster in the conserved DNA binding domain and disrupt MEF2C DNA binding. DNA binding-deficient global Mef2c heterozygous mice (Mef2c-Het) displayed numerous MCHS-related behaviors, including autism-related behaviors, changes in cortical gene expression, and deficits in cortical excitatory synaptic transmission. We detected hundreds of dysregulated genes in Mef2c-Het cortex, including significant enrichments of autism risk and excitatory neuron genes. In addition, we observed an enrichment of upregulated microglial genes, but this was not due to neuroinflammation in the Mef2c-Het cortex. Importantly, conditional Mef2c heterozygosity in forebrain excitatory neurons reproduced a subset of the Mef2c-Het phenotypes, while conditional Mef2c heterozygosity in microglia reproduced social deficits and repetitive behavior. Conclusions: Taken together, our findings show that mutations found in individuals with MCHS disrupt the DNA-binding function of MEF2C, and DNA binding-deficient Mef2c global heterozygous mice display numerous MCHS-related phenotypes, including excitatory neuron and microglia gene expression changes. Our findings suggest that MEF2C regulates typical brain development and function through multiple cell types, including excitatory neuronal and neuroimmune populations.
MeSH term(s)	Animals ; Disease Models, Animal ; Haploinsufficiency ; MEF2 Transcription Factors/genetics ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Neurons ; Synaptic Transmission
Chemical Substances	MEF2 Transcription Factors ; Mef2c protein, mouse
Language	English
Publishing date	2020-03-31
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	209434-4
ISSN	1873-2402 ; 0006-3223
ISSN (online)	1873-2402
ISSN	0006-3223
DOI	10.1016/j.biopsych.2020.03.011
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Article: Dynamics of Arabidopsis dynamin-related protein 1C and a clathrin light chain at the plasma membrane.

Konopka, Catherine A / Backues, Steven K / Bednarek, Sebastian Y

The Plant cell

2008 Volume 20, Issue 5, Page(s) 1363–1380

Abstract: Plant morphogenesis depends on polarized exocytic and endocytic membrane trafficking. Members of the Arabidopsis thaliana dynamin-related protein 1 (DRP1) subfamily are required for polarized cell expansion and cytokinesis. Using a combination of live- ... ...

Abstract	Plant morphogenesis depends on polarized exocytic and endocytic membrane trafficking. Members of the Arabidopsis thaliana dynamin-related protein 1 (DRP1) subfamily are required for polarized cell expansion and cytokinesis. Using a combination of live-cell imaging techniques, we show that a functional DRP1C green fluorescent fusion protein (DRP1C-GFP) was localized at the division plane in dividing cells and to the plasma membrane in expanding interphase cells. In both tip growing root hairs and diffuse-polar expanding epidermal cells, DRP1C-GFP organized into dynamic foci at the cell cortex, which colocalized with a clathrin light chain fluorescent fusion protein (CLC-FFP), suggesting that DRP1C may participate in clathrin-mediated membrane dynamics. DRP1C-GFP and CLC-GFP foci dynamics are dependent on cytoskeleton organization, cytoplasmic streaming, and functional clathrin-mediated endocytic traffic. Our studies provide insight into DRP1 and clathrin dynamics in the plant cell cortex and indicate that the clathrin endocytic machinery in plants has both similarities and striking differences to that in mammalian cells and yeast.
MeSH term(s)	Arabidopsis/genetics ; Arabidopsis/metabolism ; Arabidopsis Proteins/genetics ; Arabidopsis Proteins/metabolism ; Cell Membrane/metabolism ; Clathrin Light Chains/genetics ; Clathrin Light Chains/metabolism ; Cytoplasm/metabolism ; Dynamins/genetics ; Dynamins/metabolism ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Immunoblotting ; Microscopy, Confocal ; Microscopy, Fluorescence ; Plant Roots/metabolism ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism
Chemical Substances	Arabidopsis Proteins ; Clathrin Light Chains ; Recombinant Proteins ; Green Fluorescent Proteins (147336-22-9) ; DRP1C protein, Arabidopsis (EC 3.6.5.5) ; Dynamins (EC 3.6.5.5)
Language	English
Publishing date	2008-05-23
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	623171-8
ISSN	1532-298X ; 1040-4651
ISSN (online)	1532-298X
ISSN	1040-4651
DOI	10.1105/tpc.108.059428
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Article: Variable-angle epifluorescence microscopy: a new way to look at protein dynamics in the plant cell cortex

Konopka, Catherine A / Bednarek, Sebastian Y

Plant journal. 2008 Jan., v. 53, no. 1

2008

Abstract	Live-cell microscopy imaging of fluorescent-tagged fusion proteins is an essential tool for cell biologists. Total internal reflection fluorescence microscopy (TIRFM) has joined confocal microscopy as a complementary system for the imaging of cell surface protein dynamics in mammalian and yeast systems because of its high temporal and spatial resolution. Here we present an alternative to TIRFM, termed variable-angle epifluorescence microscopy (VAEM), for the visualization of protein dynamics at or near the plasma membrane of plant epidermal cells and root hairs in whole, intact seedlings that provides high-signal, low-background and near real-time imaging. VAEM uses highly oblique subcritical incident angles to decrease background fluorophore excitation. We discuss the utilities and advantages of VAEM for imaging of fluorescent fusion-tagged marker proteins in studying cortical cytoskeletal and membrane proteins. We believe that the application of VAEM will be an invaluable imaging tool for plant cell biologists.
Keywords	cortex ; cytoskeletal proteins ; fluorescence microscopy ; image analysis ; membrane proteins ; plant proteins ; plasma membrane ; root hairs ; seedlings ; surface proteins ; yeasts
Language	English
Dates of publication	2008-01
Size	p. 186-196.
Publisher	Blackwell Publishing Ltd
Publishing place	Oxford, UK
Document type	Article
ZDB-ID	1088037-9
ISSN	1365-313X ; 0960-7412
ISSN (online)	1365-313X
ISSN	0960-7412
DOI	10.1111/j.1365-313X.2007.03306.x
Database	NAL-Catalogue (AGRICOLA)

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Article: Comparison of the Dynamics and Functional Redundancy of the Arabidopsis Dynamin-Related Isoforms DRP1A and DRP1C during Plant Development

Konopka, Catherine A / Bednarek, Sebastian Y

Plant physiology. 2008 Aug., v. 147, no. 4

2008

Abstract	Members of the Arabidopsis (Arabidopsis thaliana) DYNAMIN-RELATED PROTEIN1 (DRP1) family are required for cytokinesis and cell expansion. Two isoforms, DRP1A and DRP1C, are required for plasma membrane maintenance during stigmatic papillae expansion and pollen development, respectively. It is unknown whether the DRP1s function interchangeably or if they have distinct roles during cell division and expansion. DRP1C was previously shown to form dynamic foci in the cell cortex, which colocalize with part of the clathrin endocytic machinery in plants. DRP1A localizes to the plasma membrane, but its cortical organization and dynamics have not been determined. Using dual color labeling with live cell imaging techniques, we showed that DRP1A also forms discreet dynamic foci in the epidermal cell cortex. Although the foci overlap with those formed by DRP1C and clathrin light chain, there are clear differences in behavior and response to pharmacological inhibitors between DRP1A and DRP1C foci. Possible functional or regulatory differences between DRP1A and DRP1C were supported by the failure of DRP1C to functionally compensate for the absence of DRP1A. Our studies indicated that the DRP1 isoforms function or are regulated differently during cell expansion.
Keywords	Arabidopsis thaliana ; clathrin ; color ; cytokinesis ; plant development ; plasma membrane ; pollen
Language	English
Dates of publication	2008-08
Size	p. 1590-1602.
Publishing place	American Society of Plant Biologists
Document type	Article
ZDB-ID	208914-2
ISSN	1532-2548 ; 0032-0889
ISSN (online)	1532-2548
ISSN	0032-0889
Database	NAL-Catalogue (AGRICOLA)

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