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Article ; Online: CHX14 is a plasma membrane K-efflux transporter that regulates K(+) redistribution in Arabidopsis thaliana.

Zhao, Jian / Li, Penghui / Motes, Christy M / Park, Sunghun / Hirschi, Kendal D

2015 Volume 38, Issue 11, Page(s) 2223–2238

Abstract: Potassium (K(+) ) is essential for plant growth and development, yet the molecular identity ... of many K(+) transporters remains elusive. Here we characterized cation/H(+) exchanger (CHX) 14 ... as a plasma membrane K(+) transporter. CHX14 expression was induced by elevated K(+) and histochemical analysis ...

Abstract	Potassium (K(+) ) is essential for plant growth and development, yet the molecular identity of many K(+) transporters remains elusive. Here we characterized cation/H(+) exchanger (CHX) 14 as a plasma membrane K(+) transporter. CHX14 expression was induced by elevated K(+) and histochemical analysis of CHX14 promoter::GUS transgenic plants indicated that CHX14 was expressed in xylem parenchyma of root and shoot vascular tissues of seedlings. CHX14 knockout (chx14) and CHX14 overexpression seedlings displayed different growth phenotypes during K(+) stress as compared with wild-type seedlings. Roots of mutant seedlings displayed higher K(+) uptake rates than wild-type roots. CHX14 expression in yeast cells deficient in K(+) uptake renders the mutant cells more sensitive to deficiencies of K(+) in the medium. CHX14 mediates K(+) efflux in yeast cells loaded with high K(+) . Uptake experiments using (86) Rb(+) as a tracer for K(+) with both yeast and plant mutants demonstrated that CHX14 expression in yeast and in planta mediated low-affinity K(+) efflux. Functional green fluorescent protein (GFP)-tagged versions of CHX14 were localized to both the yeast and plant plasma membranes. Taken together, we suggest that CHX14 is a plasma membrane K(+) efflux transporter involved in K(+) homeostasis and K(+) recirculation.
MeSH term(s)	Arabidopsis/genetics ; Arabidopsis/metabolism ; Arabidopsis Proteins/analysis ; Arabidopsis Proteins/genetics ; Arabidopsis Proteins/physiology ; Cell Membrane/metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Plant ; Homeostasis ; Plant Roots/genetics ; Plant Roots/metabolism ; Plant Shoots/genetics ; Plant Shoots/metabolism ; Potassium/metabolism ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Seedlings/genetics ; Seedlings/metabolism ; Sodium-Hydrogen Exchangers/analysis ; Sodium-Hydrogen Exchangers/genetics ; Sodium-Hydrogen Exchangers/physiology
Chemical Substances	Arabidopsis Proteins ; CHX17 protein, Arabidopsis ; Sodium-Hydrogen Exchangers ; Potassium (RWP5GA015D)
Language	English
Publishing date	2015-11
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	391893-2
ISSN	1365-3040 ; 0140-7791
ISSN (online)	1365-3040
ISSN	0140-7791
DOI	10.1111/pce.12524
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Article: CHX14 is a plasma membrane K‐efflux transporter that regulates K+ redistribution in Arabidopsis thaliana

Zhao, Jian / Hirschi, Kendal D / Li, Penghui / Motes, Christy M / Park, Sunghun

Plant, cell and environment. 2015 Nov., v. 38, no. 11

2015

Abstract: Potassium (K+) is essential for plant growth and development, yet the molecular identity of many K+ ... transporters remains elusive. Here we characterized cation/H+ exchanger (CHX) 14 as a plasma membrane K+ ... transporter. CHX14 expression was induced by elevated K+ and histochemical analysis of CHX14 promoter::GUS ...

Abstract	Potassium (K+) is essential for plant growth and development, yet the molecular identity of many K+ transporters remains elusive. Here we characterized cation/H+ exchanger (CHX) 14 as a plasma membrane K+ transporter. CHX14 expression was induced by elevated K+ and histochemical analysis of CHX14 promoter::GUS transgenic plants indicated that CHX14 was expressed in xylem parenchyma of root and shoot vascular tissues of seedlings. CHX14 knockout (chx14) and CHX14 overexpression seedlings displayed different growth phenotypes during K+ stress as compared with wild‐type seedlings. Roots of mutant seedlings displayed higher K+ uptake rates than wild‐type roots. CHX14 expression in yeast cells deficient in K+ uptake renders the mutant cells more sensitive to deficiencies of K+ in the medium. CHX14 mediates K+ efflux in yeast cells loaded with high K+. Uptake experiments using 86Rb+ as a tracer for K+ with both yeast and plant mutants demonstrated that CHX14 expression in yeast and in planta mediated low‐affinity K+ efflux. Functional green fluorescent protein (GFP)‐tagged versions of CHX14 were localized to both the yeast and plant plasma membranes. Taken together, we suggest that CHX14 is a plasma membrane K+ efflux transporter involved in K+ homeostasis and K+ recirculation.
Keywords	Arabidopsis thaliana ; gene overexpression ; green fluorescent protein ; homeostasis ; mutants ; parenchyma (plant tissue) ; phenotype ; plant growth ; plasma membrane ; potassium ; protons ; roots ; seedlings ; shoots ; transgenic plants ; transporters ; vascular tissues ; yeasts
Language	English
Dates of publication	2015-11
Size	p. 2223-2238.
Publishing place	Blackwell Scientific Publications
Document type	Article
ZDB-ID	391893-2
ISSN	1365-3040 ; 0140-7791
ISSN (online)	1365-3040
ISSN	0140-7791
DOI	10.1111/pce.12524
Database	NAL-Catalogue (AGRICOLA)

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Article: Bioavailability of transgenic microRNAs in genetically modified plants

hirschi, kendal

Genes & nutrition, 12:17

2017

Abstract: BACKGROUND: Transgenic expression of small RNAs is a prevalent approach in agrobiotechnology for the global enhancement of plant foods. Meanwhile, emerging studies have, on the one hand, emphasized the potential of transgenic microRNAs (miRNAs) as novel ... ...

Abstract	BACKGROUND: Transgenic expression of small RNAs is a prevalent approach in agrobiotechnology for the global enhancement of plant foods. Meanwhile, emerging studies have, on the one hand, emphasized the potential of transgenic microRNAs (miRNAs) as novel dietary therapeutics and, on the other, suggested potential food safety issues if harmful miRNAs are absorbed and bioactive. For these reasons, it is necessary to evaluate the bioavailability of transgenic miRNAs in genetically modified crops. RESULTS: As a pilot study, two transgenic Arabidopsis lines ectopically expressing unique miRNAs were compared and contrasted with the plant bioavailable small RNA MIR2911 for digestive stability and serum bioavailability. The expression levels of these transgenic miRNAs in Arabidopsis were found to be comparable to that of MIR2911 in fresh tissues. Assays of digestive stability in vitro and in vivo suggested the transgenic miRNAs and MIR2911 had comparable resistance to degradation. Healthy mice consuming diets rich in Arabidopsis lines expressing these miRNAs displayed MIR2911 in the bloodstream but no detectable levels of the transgenic miRNAs. CONCLUSIONS: These preliminary results imply digestive stability and high expression levels of miRNAs in plants do not readily equate to bioavailability. This initial work suggests novel engineering strategies be employed to enhance miRNA bioavailability when attempting to use transgenic foods as a delivery platform.
Keywords	Bioavailability ; Genetically modified organisms ; Dietary microRNAs ; Digestive stability ; MIR2911 ; Mice
Language	English
Document type	Article
Database	Repository for Life Sciences

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Article: AtCHX13 is a plasma membrane K+ transporter.

Zhao, Jian / Cheng, Ning-Hui / Motes, Christy M / Blancaflor, Elison B / Moore, Miranda / Gonzales, Naomi / Padmanaban, Senthilkumar / Sze, Heven / Ward, John M / Hirschi, Kendal D

Plant physiology

2008 Volume 148, Issue 2, Page(s) 796–807

Abstract: ... demonstrated that AtCHX13 mediated high-affinity K+ uptake in yeast and in plant cells with a K(m) of 136 and ... Potassium (K+) homeostasis is essential for diverse cellular processes, although how various cation ... transporters collaborate to maintain a suitable K+ required for growth and development is poorly understood ...

Abstract	Potassium (K+) homeostasis is essential for diverse cellular processes, although how various cation transporters collaborate to maintain a suitable K+ required for growth and development is poorly understood. The Arabidopsis (Arabidopsis thaliana) genome contains numerous cation:proton antiporters (CHX), which may mediate K+ transport; however, the vast majority of these transporters remain uncharacterized. Here, we show that AtCHX13 (At2g30240) has a role in K+ acquisition. AtCHX13 suppressed the sensitivity of yeast (Saccharomyces cerevisiae) mutant cells defective in K+ uptake. Uptake experiments using (86)Rb+ as a tracer for K+ demonstrated that AtCHX13 mediated high-affinity K+ uptake in yeast and in plant cells with a K(m) of 136 and 196 microm, respectively. Functional green fluorescent protein-tagged versions localized to the plasma membrane of both yeast and plant. Seedlings of null chx13 mutants were sensitive to K+ deficiency conditions, whereas overexpression of AtCHX13 reduced the sensitivity to K+ deficiency. Collectively, these results suggest that AtCHX13 mediates relatively high-affinity K+ uptake, although the mode of transport is unclear at present. AtCHX13 expression is induced in roots during K+-deficient conditions. These results indicate that one role of AtCHX13 is to promote K+ uptake into plants when K+ is limiting in the environment.
MeSH term(s)	Arabidopsis/genetics ; Arabidopsis/metabolism ; Arabidopsis Proteins/genetics ; Arabidopsis Proteins/metabolism ; Biological Transport ; Cation Transport Proteins/genetics ; Cation Transport Proteins/metabolism ; Cell Membrane/metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Phenotype ; Plant Roots/genetics ; Plant Roots/metabolism ; Plasmids ; Potassium/metabolism ; RNA, Plant/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism
Chemical Substances	Arabidopsis Proteins ; Cation Transport Proteins ; RNA, Plant ; Potassium (RWP5GA015D)
Language	English
Publishing date	2008-08-01
Publishing country	United States
Document type	Journal Article ; 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.108.124248
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Article: AtCCX3 is an Arabidopsis endomembrane H+ -dependent K+ transporter.

Morris, Jay / Tian, Hui / Park, Sunghun / Sreevidya, Coimbatore S / Ward, John M / Hirschi, Kendal D

Plant physiology

2008 Volume 148, Issue 3, Page(s) 1474–1486

Abstract: ... that Arabidopsis AtCCX3 (At3g14070) and AtCCX4 (At1g54115) can suppress yeast mutants defective in Na(+), K(+), and ... cells by excess Na(+), K(+), and Mn(2+). Functional epitope-tagged AtCCX3 fusion proteins were localized ... that AtCCX3 is an endomembrane-localized H(+)-dependent K(+) transporter with apparent Na(+) and Mn(2+ ...

Abstract	The Arabidopsis (Arabidopsis thaliana) cation calcium exchangers (CCXs) were recently identified as a subfamily of cation transporters; however, no plant CCXs have been functionally characterized. Here, we show that Arabidopsis AtCCX3 (At3g14070) and AtCCX4 (At1g54115) can suppress yeast mutants defective in Na(+), K(+), and Mn(2+) transport. We also report high-capacity uptake of (86)Rb(+) in tonoplast-enriched vesicles from yeast expressing AtCCX3. Cation competition studies showed inhibition of (86)Rb(+) uptake in AtCCX3 cells by excess Na(+), K(+), and Mn(2+). Functional epitope-tagged AtCCX3 fusion proteins were localized to endomembranes in plants and yeast. In Arabidopsis, AtCCX3 is primarily expressed in flowers, while AtCCX4 is expressed throughout the plant. Quantitative polymerase chain reaction showed that expression of AtCCX3 increased in plants treated with NaCl, KCl, and MnCl(2). Insertional mutant lines of AtCCX3 and AtCCX4 displayed no apparent growth defects; however, overexpression of AtCCX3 caused increased Na(+) accumulation and increased (86)Rb(+) transport. Uptake of (86)Rb(+) increased in tonoplast-enriched membranes isolated from Arabidopsis lines expressing CCX3 driven by the cauliflower mosaic virus 35S promoter. Overexpression of AtCCX3 in tobacco (Nicotiana tabacum) produced lesions in the leaves, stunted growth, and resulted in the accumulation of higher levels of numerous cations. In summary, these findings suggest that AtCCX3 is an endomembrane-localized H(+)-dependent K(+) transporter with apparent Na(+) and Mn(2+) transport properties distinct from those of previously characterized plant transporters.
MeSH term(s)	Arabidopsis/genetics ; Arabidopsis/metabolism ; Ion Transport ; Membrane Transport Proteins/genetics ; Membrane Transport Proteins/metabolism ; Polymerase Chain Reaction ; Potassium/metabolism ; Protons
Chemical Substances	Membrane Transport Proteins ; Protons ; Potassium (RWP5GA015D)
Language	English
Publishing date	2008-09-05
Publishing country	United States
Document type	Journal Article ; 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.108.118810
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Article: AtCHX13 Is a Plasma Membrane K+ Transporter

Zhao, Jian / Cheng, Ning-Hui / Motes, Christy M / Blancaflor, Elison B / Moore, Miranda / Gonzales, Naomi / Padmanaban, Senthilkumar / Sze, Heven / Ward, John M / Hirschi, Kendal D

Plant physiology. 2008 Oct., v. 148, no. 2

2008

Abstract: Potassium (K+) homeostasis is essential for diverse cellular processes, although how various cation ... transporters collaborate to maintain a suitable K+ required for growth and development is poorly understood ... K+ transport; however, the vast majority of these transporters remain uncharacterized. Here, we show ...

Abstract	Potassium (K+) homeostasis is essential for diverse cellular processes, although how various cation transporters collaborate to maintain a suitable K+ required for growth and development is poorly understood. The Arabidopsis (Arabidopsis thaliana) genome contains numerous cation:proton antiporters (CHX), which may mediate K+ transport; however, the vast majority of these transporters remain uncharacterized. Here, we show that AtCHX13 (At2g30240) has a role in K+ acquisition. AtCHX13 suppressed the sensitivity of yeast (Saccharomyces cerevisiae) mutant cells defective in K+ uptake. Uptake experiments using 86Rb+ as a tracer for K+ demonstrated that AtCHX13 mediated high-affinity K+ uptake in yeast and in plant cells with a Km of 136 and 196 μM, respectively. Functional green fluorescent protein-tagged versions localized to the plasma membrane of both yeast and plant. Seedlings of null chx13 mutants were sensitive to K+ deficiency conditions, whereas overexpression of AtCHX13 reduced the sensitivity to K+ deficiency. Collectively, these results suggest that AtCHX13 mediates relatively high-affinity K+ uptake, although the mode of transport is unclear at present. AtCHX13 expression is induced in roots during K+-deficient conditions. These results indicate that one role of AtCHX13 is to promote K+ uptake into plants when K+ is limiting in the environment.
Keywords	Arabidopsis thaliana ; Saccharomyces cerevisiae ; antiporters ; cations ; genome ; homeostasis ; labeling techniques ; mutants ; plasma membrane ; potassium ; roots ; seedlings ; yeasts
Language	English
Dates of publication	2008-10
Size	p. 796-807.
Publishing place	American Society of Plant Biologists
Document type	Article
Note	Includes references
ZDB-ID	208914-2
ISSN	1532-2548 ; 0032-0889
ISSN (online)	1532-2548
ISSN	0032-0889
DOI	10.1104/pp.108.124248
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Endothelial Cell Differentiation and Hemogenic Specification.

Aragon, Jordon W / Hirschi, Karen K

Cold Spring Harbor perspectives in medicine

2022 Volume 12, Issue 7

Abstract: Formation of the vasculature is a critical step within the developing embryo and its disruption causes early embryonic lethality. This complex process is driven by a cascade of signaling events that controls differentiation of mesodermal progenitors into ...

Abstract	Formation of the vasculature is a critical step within the developing embryo and its disruption causes early embryonic lethality. This complex process is driven by a cascade of signaling events that controls differentiation of mesodermal progenitors into primordial endothelial cells and their further specification into distinct subtypes (arterial, venous, hemogenic) that are needed to generate a blood circulatory network. Hemogenic endothelial cells give rise to hematopoietic stem and progenitor cells that generate all blood cells in the body during embryogenesis and postnatally. We focus our discussion on the regulation of endothelial cell differentiation, and subsequent hemogenic specification, and highlight many of the signaling pathways involved in these processes, which are conserved across vertebrates. Gaining a better understanding of the regulation of these processes will yield insights needed to optimize the treatment of vascular and hematopoietic disease and generate human stem cell-derived vascular and hematopoietic cells for tissue engineering and regenerative medicine.
MeSH term(s)	Animals ; Cell Differentiation/physiology ; Hemangioblasts ; Hematopoiesis/physiology ; Hematopoietic Stem Cells/metabolism ; Humans ; Signal Transduction/physiology
Language	English
Publishing date	2022-07-29
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural
ISSN	2157-1422
ISSN (online)	2157-1422
DOI	10.1101/cshperspect.a041164
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Article: AtCCX3 Is an Arabidopsis Endomembrane H⁺-Dependent K⁺ Transporter

Morris, Jay / Tian, Hui / Park, Sunghun / Sreevidya, Coimbatore S / Ward, John M / Hirschi, Kendal D

Plant physiology. 2008 Nov., v. 148, no. 3

2008

Abstract: ... that Arabidopsis AtCCX3 (At3g14070) and AtCCX4 (At1g54115) can suppress yeast mutants defective in Na⁺, K⁺, and ... K⁺, and Mn²⁺. Functional epitope-tagged AtCCX3 fusion proteins were localized to endomembranes ... localized H⁺-dependent K⁺ transporter with apparent Na⁺ and Mn²⁺ transport properties distinct ...

Abstract	The Arabidopsis (Arabidopsis thaliana) cation calcium exchangers (CCXs) were recently identified as a subfamily of cation transporters; however, no plant CCXs have been functionally characterized. Here, we show that Arabidopsis AtCCX3 (At3g14070) and AtCCX4 (At1g54115) can suppress yeast mutants defective in Na⁺, K⁺, and Mn²⁺ transport. We also report high-capacity uptake of ⁸⁶Rb⁺ in tonoplast-enriched vesicles from yeast expressing AtCCX3. Cation competition studies showed inhibition of ⁸⁶Rb⁺ uptake in AtCCX3 cells by excess Na⁺, K⁺, and Mn²⁺. Functional epitope-tagged AtCCX3 fusion proteins were localized to endomembranes in plants and yeast. In Arabidopsis, AtCCX3 is primarily expressed in flowers, while AtCCX4 is expressed throughout the plant. Quantitative polymerase chain reaction showed that expression of AtCCX3 increased in plants treated with NaCl, KCl, and MnCl₂. Insertional mutant lines of AtCCX3 and AtCCX4 displayed no apparent growth defects; however, overexpression of AtCCX3 caused increased Na⁺ accumulation and increased ⁸⁶Rb⁺ transport. Uptake of ⁸⁶Rb⁺ increased in tonoplast-enriched membranes isolated from Arabidopsis lines expressing CCX3 driven by the cauliflower mosaic virus 35S promoter. Overexpression of AtCCX3 in tobacco (Nicotiana tabacum) produced lesions in the leaves, stunted growth, and resulted in the accumulation of higher levels of numerous cations. In summary, these findings suggest that AtCCX3 is an endomembrane-localized H⁺-dependent K⁺ transporter with apparent Na⁺ and Mn²⁺ transport properties distinct from those of previously characterized plant transporters.
Keywords	Arabidopsis thaliana ; antiporters ; potassium ; gene expression ; yeasts ; gene overexpression ; Nicotiana tabacum ; tobacco ; transgenic plants ; plant growth
Language	English
Dates of publication	2008-11
Size	p. 1474-1486.
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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Article ; Online: Understanding neural stem cell regulation

Genet, Nafiisha / Hirschi, Karen K

Regenerative medicine

2021 Volume 16, Issue 9, Page(s) 861–870

Abstract: The use of neural stem cell (NSC) therapy for the treatment of stroke patients is successfully paving its way into advanced phases of large-scale clinical trials. To understand how to optimize NSC therapeutic approaches, it is fundamental to decipher the ...

Abstract	The use of neural stem cell (NSC) therapy for the treatment of stroke patients is successfully paving its way into advanced phases of large-scale clinical trials. To understand how to optimize NSC therapeutic approaches, it is fundamental to decipher the crosstalk between NSC and other cells that comprise the NSC microenvironment (niche) and regulate their function,
MeSH term(s)	Endothelial Cells ; Humans ; Neural Stem Cells ; Neurogenesis ; Stem Cell Niche ; Stem Cell Transplantation ; Stroke/therapy
Language	English
Publishing date	2021-09-09
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2274500-2
ISSN	1746-076X ; 1746-0751
ISSN (online)	1746-076X
ISSN	1746-0751
DOI	10.2217/rme-2021-0022
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Article: Tissue-Resident Macrophage Development and Function.

Wu, Yinyu / Hirschi, Karen K

Frontiers in cell and developmental biology

2021 Volume 8, Page(s) 617879

Abstract: Tissue-resident macrophages have been associated with important and diverse biological processes such as native immunity, tissue homeostasis and angiogenesis during development and postnatally. Thus, it is critical to understand the origins and functions ...

Abstract	Tissue-resident macrophages have been associated with important and diverse biological processes such as native immunity, tissue homeostasis and angiogenesis during development and postnatally. Thus, it is critical to understand the origins and functions of tissue-resident macrophages, as well as mechanisms underlying their regulation. It is now well accepted that murine macrophages are produced during three consecutive waves of hematopoietic development. The first wave of macrophage formation takes place during primitive hematopoiesis, which occurs in the yolk sac, and gives rise to primitive erythroid, megakaryocyte and macrophage progenitors. These "primitive" macrophage progenitors ultimately give rise to microglia in the adult brain. The second wave, which also occurs in the yolk sac, generates multipotent erythro-myeloid progenitors (EMP), which give rise to tissue-resident macrophages. Tissue-resident macrophages derived from EMP reside in diverse niches of different tissues except the brain, and demonstrate tissue-specific functions therein. The third wave of macrophages derives from hematopoietic stem cells (HSC) that are formed in the aorta-gonad-mesonephros (AGM) region of the embryo and migrate to, and colonize, the fetal liver. These HSC-derived macrophages are a long-lived pool that will last throughout adulthood. In this review, we discuss the developmental origins of tissue-resident macrophages, their molecular regulation in specific tissues, and their impact on embryonic development and postnatal homeostasis.
Language	English
Publishing date	2021-01-08
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2020.617879
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