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  1. Article ; Online: Using Fluorescence Recovery After Photobleaching to Study Gap Junctional Communication In Vitro.

    Kuzma-Kuzniarska, Maria / Yapp, Clarence / Hulley, Philippa A

    Methods in molecular biology (Clifton, N.J.)

    2016  Volume 1437, Page(s) 171–179

    Abstract: Fluorescence recovery after photobleaching (FRAP) is a microscopy-based technique to study the movement of fluorescent molecules inside a cell. Although initially developed to investigate intracellular mobility, FRAP can be also used to measure ... ...

    Abstract Fluorescence recovery after photobleaching (FRAP) is a microscopy-based technique to study the movement of fluorescent molecules inside a cell. Although initially developed to investigate intracellular mobility, FRAP can be also used to measure intercellular dynamics. This chapter describes how to perform FRAP experiment to study gap junctional communication in living cells. The procedures described here can be carried out with a laser-scanning confocal microscope and any in vitro cultured cells known to communicate via gap junctions. In addition, the method can be easily adjusted to measure gap junction function in 3D cell cultures as well as ex vivo tissue.
    Language English
    Publishing date 2016
    Publishing country United States
    Document type Journal Article
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-3664-9_12
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  2. Article ; Online: Hnf1b haploinsufficiency differentially affects developmental target genes in a new renal cysts and diabetes mouse model.

    Niborski, Leticia L / Paces-Fessy, Mélanie / Ricci, Pierbruno / Bourgeois, Adeline / Magalhães, Pedro / Kuzma-Kuzniarska, Maria / Lesaulnier, Celine / Reczko, Martin / Declercq, Edwige / Zürbig, Petra / Doucet, Alain / Umbhauer, Muriel / Cereghini, Silvia

    Disease models & mechanisms

    2021  Volume 14, Issue 5

    Abstract: Heterozygous mutations in HNF1B cause the complex syndrome renal cysts and diabetes (RCAD), characterized by developmental abnormalities of the kidneys, genital tracts and pancreas, and a variety of renal, pancreas and liver dysfunctions. The ... ...

    Abstract Heterozygous mutations in HNF1B cause the complex syndrome renal cysts and diabetes (RCAD), characterized by developmental abnormalities of the kidneys, genital tracts and pancreas, and a variety of renal, pancreas and liver dysfunctions. The pathogenesis underlying this syndrome remains unclear as mice with heterozygous null mutations have no phenotype, while constitutive/conditional Hnf1b ablation leads to more severe phenotypes. We generated a novel mouse model carrying an identified human mutation at the intron-2 splice donor site. Unlike heterozygous mice previously characterized, mice heterozygous for the splicing mutation exhibited decreased HNF1B protein levels and bilateral renal cysts from embryonic day 15, originated from glomeruli, early proximal tubules (PTs) and intermediate nephron segments, concurrently with delayed PT differentiation, hydronephrosis and rare genital tract anomalies. Consistently, mRNA sequencing showed that most downregulated genes in embryonic kidneys were primarily expressed in early PTs and the loop of Henle and involved in ion/drug transport, organic acid and lipid metabolic processes, while the expression of previously identified targets upon Hnf1b ablation, including cystic disease genes, was weakly or not affected. Postnatal analyses revealed renal abnormalities, ranging from glomerular cysts to hydronephrosis and, rarely, multicystic dysplasia. Urinary proteomics uncovered a particular profile predictive of progressive decline in kidney function and fibrosis, and displayed common features with a recently reported urine proteome in an RCAD pediatric cohort. Altogether, our results show that reduced HNF1B levels lead to developmental disease phenotypes associated with the deregulation of a subset of HNF1B targets. They further suggest that this model represents a unique clinical/pathological viable model of the RCAD disease.
    MeSH term(s) Animals ; Animals, Newborn ; Cell Polarity ; Central Nervous System Diseases/genetics ; Central Nervous System Diseases/pathology ; Cilia/pathology ; Dental Enamel/abnormalities ; Dental Enamel/pathology ; Diabetes Mellitus, Experimental/genetics ; Diabetes Mellitus, Experimental/pathology ; Diabetes Mellitus, Type 2/genetics ; Diabetes Mellitus, Type 2/pathology ; Disease Models, Animal ; Embryo, Mammalian/pathology ; Gene Dosage ; Gene Expression Profiling ; Genes, Developmental ; Haploinsufficiency/genetics ; Hepatocyte Nuclear Factor 1-beta/genetics ; Heterozygote ; Humans ; Hydronephrosis/complications ; Kidney Diseases, Cystic/genetics ; Kidney Diseases, Cystic/pathology ; Kidney Glomerulus/pathology ; Kidney Tubules/pathology ; Mice, Inbred C57BL ; Mutation/genetics ; Nephrons/pathology ; RNA Splicing/genetics ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Severity of Illness Index ; Mice
    Chemical Substances HNF1B protein, human ; Hnf1b protein, mouse ; RNA, Messenger ; Hepatocyte Nuclear Factor 1-beta (138674-15-4)
    Language English
    Publishing date 2021-05-04
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2451104-3
    ISSN 1754-8411 ; 1754-8403
    ISSN (online) 1754-8411
    ISSN 1754-8403
    DOI 10.1242/dmm.047498
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Lovastatin-Mediated Changes in Human Tendon Cells.

    Kuzma-Kuzniarska, Maria / Cornell, Hannah R / Moneke, Michael C / Carr, Andrew J / Hulley, Philippa A

    Journal of cellular physiology

    2015  Volume 230, Issue 10, Page(s) 2543–2551

    Abstract: Statins are among the most widely prescribed drugs worldwide. Numerous studies have shown their beneficial effects in prevention of cardiovascular disease through cholesterol-lowering and anti-atherosclerotic properties. Although some statin patients may ...

    Abstract Statins are among the most widely prescribed drugs worldwide. Numerous studies have shown their beneficial effects in prevention of cardiovascular disease through cholesterol-lowering and anti-atherosclerotic properties. Although some statin patients may experience muscle-related symptoms, severe side effects of statin therapy are rare, primarily due to extensive first-pass metabolism in the liver. Skeletal muscles appear to be the main site of side effects; however, recently some statin-related adverse effects have been described in tendon. The mechanism behind these side effects remains unknown. This is the first study that explores tendon-specific effects of statins in human primary tenocytes. The cells were cultured with different concentrations of lovastatin for up to 1 week. No changes in cell viability or morphology were observed in tenocytes incubated with therapeutic doses. Short-term exposure to lovastatin concentrations outside the therapeutic range had no effect on tenocyte viability; however, cell migration was reduced. Simvastatin and atorvastatin, two other drug family members, also reduced the migratory properties of the cells. Prolonged exposure to high concentrations of lovastatin induced changes in cytoskeleton leading to cell rounding and decreased levels of mRNA for matrix proteins, but increased BMP-2 expression. Gap junctional communication was impaired but due to cell shape change and separation rather than direct gap junction inhibition. These effects were accompanied by inhibition of prenylation of Rap1a small GTPase. Collectively, we showed that statins in a dose-dependent manner decrease migration of human tendon cells, alter their expression profile and impair the functional network, but do not inhibit gap junction function.
    MeSH term(s) Atorvastatin ; Cholesterol/metabolism ; Cytoskeleton/drug effects ; Cytoskeleton/metabolism ; Gap Junctions/drug effects ; Heptanoic Acids/pharmacology ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology ; Lovastatin/pharmacology ; Pyrroles/pharmacology ; Simvastatin/pharmacology ; Tendons/drug effects ; Tendons/metabolism
    Chemical Substances Heptanoic Acids ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; Pyrroles ; Cholesterol (97C5T2UQ7J) ; Lovastatin (9LHU78OQFD) ; Atorvastatin (A0JWA85V8F) ; Simvastatin (AGG2FN16EV)
    Language English
    Publishing date 2015-04-28
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3116-1
    ISSN 1097-4652 ; 0021-9541
    ISSN (online) 1097-4652
    ISSN 0021-9541
    DOI 10.1002/jcp.25010
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    In stock of ZB MED Cologne/Königswinter

    Uc I Zs.212: Show issues Location:
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    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

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  4. Article ; Online: Hnf1b haploinsufficiency differentially affects developmental target genes in a new renal cysts and diabetes mouse model

    Leticia L. Niborski / Mélanie Paces-Fessy / Pierbruno Ricci / Adeline Bourgeois / Pedro Magalhães / Maria Kuzma-Kuzniarska / Celine Lesaulnier / Martin Reczko / Edwige Declercq / Petra Zürbig / Alain Doucet / Muriel Umbhauer / Silvia Cereghini

    Disease Models & Mechanisms, Vol 14, Iss

    2021  Volume 5

    Abstract: Heterozygous mutations in HNF1B cause the complex syndrome renal cysts and diabetes (RCAD), characterized by developmental abnormalities of the kidneys, genital tracts and pancreas, and a variety of renal, pancreas and liver dysfunctions. The ... ...

    Abstract Heterozygous mutations in HNF1B cause the complex syndrome renal cysts and diabetes (RCAD), characterized by developmental abnormalities of the kidneys, genital tracts and pancreas, and a variety of renal, pancreas and liver dysfunctions. The pathogenesis underlying this syndrome remains unclear as mice with heterozygous null mutations have no phenotype, while constitutive/conditional Hnf1b ablation leads to more severe phenotypes. We generated a novel mouse model carrying an identified human mutation at the intron-2 splice donor site. Unlike heterozygous mice previously characterized, mice heterozygous for the splicing mutation exhibited decreased HNF1B protein levels and bilateral renal cysts from embryonic day 15, originated from glomeruli, early proximal tubules (PTs) and intermediate nephron segments, concurrently with delayed PT differentiation, hydronephrosis and rare genital tract anomalies. Consistently, mRNA sequencing showed that most downregulated genes in embryonic kidneys were primarily expressed in early PTs and the loop of Henle and involved in ion/drug transport, organic acid and lipid metabolic processes, while the expression of previously identified targets upon Hnf1b ablation, including cystic disease genes, was weakly or not affected. Postnatal analyses revealed renal abnormalities, ranging from glomerular cysts to hydronephrosis and, rarely, multicystic dysplasia. Urinary proteomics uncovered a particular profile predictive of progressive decline in kidney function and fibrosis, and displayed common features with a recently reported urine proteome in an RCAD pediatric cohort. Altogether, our results show that reduced HNF1B levels lead to developmental disease phenotypes associated with the deregulation of a subset of HNF1B targets. They further suggest that this model represents a unique clinical/pathological viable model of the RCAD disease.
    Keywords hnf1b transcription factor ; rcad syndrome ; gene dosage ; glomerular and proximal tubule cysts ; mouse models ; transcriptomics ; Medicine ; R ; Pathology ; RB1-214
    Subject code 616
    Language English
    Publishing date 2021-05-01T00:00:00Z
    Publisher The Company of Biologists
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching.

    Kuzma-Kuzniarska, Maria / Yapp, Clarence / Pearson-Jones, Thomas W / Jones, Andrew K / Hulley, Philippa A

    Journal of biomedical optics

    2014  Volume 19, Issue 1, Page(s) 15001

    Abstract: Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. ... ...

    Abstract Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both 18 β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures.
    MeSH term(s) Adult ; Carbenoxolone/chemistry ; Cell Communication ; Cell Culture Techniques ; Cells, Cultured ; Collagen/chemistry ; Diffusion ; Female ; Fluorescence Recovery After Photobleaching/methods ; Gap Junctions/pathology ; Glycyrrhetinic Acid/analogs & derivatives ; Glycyrrhetinic Acid/chemistry ; HeLa Cells ; Humans ; Imaging, Three-Dimensional ; Immunohistochemistry ; Male ; Middle Aged ; Tendons/cytology ; Tendons/pathology
    Chemical Substances 18alpha-glycyrrhetinic acid (1449-05-4) ; Collagen (9007-34-5) ; Carbenoxolone (MM6384NG73) ; Glycyrrhetinic Acid (P540XA09DR)
    Language English
    Publishing date 2014-01-05
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1309154-2
    ISSN 1560-2281 ; 1083-3668
    ISSN (online) 1560-2281
    ISSN 1083-3668
    DOI 10.1117/1.JBO.19.1.015001
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 4699: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (2.OG)
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  6. Article ; Online: Integration potential of mouse and human bone marrow-derived mesenchymal stem cells.

    Kuzma-Kuzniarska, Maria / Rak-Raszewska, Aleksandra / Kenny, Simon / Edgar, David / Wilm, Bettina / Fuente Mora, Cristina / Davies, Jamie A / Murray, Patricia

    Differentiation; research in biological diversity

    2012  Volume 83, Issue 3, Page(s) 128–137

    Abstract: Mesenchymal stem cells (MSCs) are a multipotent cell population which has been described to exert renoprotective and regenerative effects in experimental models of kidney injury. Several lines of evidence indicate that MSCs also have the ability to ... ...

    Abstract Mesenchymal stem cells (MSCs) are a multipotent cell population which has been described to exert renoprotective and regenerative effects in experimental models of kidney injury. Several lines of evidence indicate that MSCs also have the ability to contribute to nephrogenesis, suggesting that the cells can be employed in stem cell-based applications aimed at de novo renal tissue generation. In this study we re-evaluate the capacity of mouse and human bone marrow-derived MSCs to contribute to the development of renal tissue using a novel method of embryonic kidney culture. Although MSCs show expression of some genes involved in renal development, their contribution to nephrogenesis is very limited in comparison to other stem cell types tested. Furthermore, we found that both mouse and human MSCs have a detrimental effect on the ex vivo development of mouse embryonic kidney, this effect being mediated through a paracrine action. Stimulation with conditioned medium from a mouse renal progenitor population increases the ability of mouse MSCs to integrate into developing renal tissue and prevents the negative effects on kidney development, but does not appear to enhance their ability to undergo nephrogenesis.
    MeSH term(s) Animals ; Bone Marrow Cells/cytology ; Cell Differentiation ; Cells, Cultured ; Culture Media, Conditioned ; Embryonic Development ; Gene Expression Regulation, Developmental ; Humans ; Kidney/cytology ; Kidney/growth & development ; Mesenchymal Stromal Cells/cytology ; Mesenchymal Stromal Cells/metabolism ; Mice ; Paracrine Communication
    Chemical Substances Culture Media, Conditioned
    Language English
    Publishing date 2012-03
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 184540-8
    ISSN 1432-0436 ; 0301-4681
    ISSN (online) 1432-0436
    ISSN 0301-4681
    DOI 10.1016/j.diff.2011.11.004
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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1170: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
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  7. Article: Integration potential of mouse and human bone marrow-derived mesenchymal stem cells

    Kuzma-Kuzniarska, Maria / Rak-Raszewska, Aleksandra / Kenny, Simon / Edgar, David / Wilm, Bettina / Fuente Mora, Cristina / Davies, Jamie A / Murray, Patricia

    Differentiation. 2012 Mar., v. 83, no. 3

    2012  

    Abstract: Mesenchymal stem cells (MSCs) are a multipotent cell population which has been described to exert renoprotective and regenerative effects in experimental models of kidney injury. Several lines of evidence indicate that MSCs also have the ability to ... ...

    Abstract Mesenchymal stem cells (MSCs) are a multipotent cell population which has been described to exert renoprotective and regenerative effects in experimental models of kidney injury. Several lines of evidence indicate that MSCs also have the ability to contribute to nephrogenesis, suggesting that the cells can be employed in stem cell-based applications aimed at de novo renal tissue generation. In this study we re-evaluate the capacity of mouse and human bone marrow-derived MSCs to contribute to the development of renal tissue using a novel method of embryonic kidney culture. Although MSCs show expression of some genes involved in renal development, their contribution to nephrogenesis is very limited in comparison to other stem cell types tested. Furthermore, we found that both mouse and human MSCs have a detrimental effect on the ex vivo development of mouse embryonic kidney, this effect being mediated through a paracrine action. Stimulation with conditioned medium from a mouse renal progenitor population increases the ability of mouse MSCs to integrate into developing renal tissue and prevents the negative effects on kidney development, but does not appear to enhance their ability to undergo nephrogenesis.
    Keywords genes ; humans ; kidneys ; methodology ; mice ; models ; population growth ; stem cells
    Language English
    Dates of publication 2012-03
    Size p. 128-137.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 184540-8
    ISSN 1432-0436 ; 0301-4681
    ISSN (online) 1432-0436
    ISSN 0301-4681
    DOI 10.1016/j.diff.2011.11.004
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Bonn / Germany

    Z 75/707: Show issues

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