FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2948469013> ?p ?o ?g. }

• W2948469013 abstract "Background: The aim of the presented thesis was to analyze the activation of the monomeric G protein RhoA in adult cardiomyocytes and further to unravel the role of the guanine nucleotide exchange factor p63RhoGEF in this context. Previous data demonstrated that RhoA is activated in neonatal cardiomyocytes by GPCR ligands like endothelin-1 (ET-1), phenylephrine (PE) and angiotensin II (AngII). So far, a contribution of the Gq/11 protein-regulated p63RhoGEF was demonstrated in the ET-1-mediated RhoA activation in neonatal cardiomyocytes. Moreover, RhoA was described to be permanently activated in pressure-overloaded hearts and p63RhoGEF to be higher expressed in the same system. This led to the hypothesis that p63RhoGEF is involved in the GPCR-dependent RhoA regulation in adult cardiomyocytes. Methods: The obtained data result from combined analysis of isolated wildtype (WT), healthy (sham) and diseased (transverse aortic constriction, TAC) adult mouse cardiomyocytes (AMCM) as well as of cholesterol-depleted AMCM (MβCD-AMCM) and of heterozygous (HET) and homozygous (KO) p63RhoGEF-knockout AMCM. p63RhoGEF-constructs were adenovirally overexpressed in WT-AMCM. The activity of RhoA was studied with the help of an antibody which specifically recognizes the conformation of active RhoA (RhoA-GTP) by immunofluorescence stainings and confocal imaging. Changes in RhoA activity were exemplarily confirmed by a biochemical binding assay. Protein distribution in AMCM was also analyzed by immunofluorescence stainings and confocal imaging, and protein expression was detected by immunoblot analysis. Quantitative analyses of protein co-localization and distribution were performed with ImageJ. Results: Independent of the investigated condition, active RhoA was predominately localized at the sarcolemma in AMCM. Application of ET-1, PE and AngII to WT- or sham-AMCM resulted in an increased level of active RhoA and a redistribution towards the costameric regions of the sarcolemma. Neither in TAC-, MβCD-, HET- nor in KO-AMCM an activation occurred in response to the GPCR ligands. In contrast, in all four types of AMCM, ET-1 and PE reduced the RhoA activity compared to control. Interestingly, in TAC-, MβCD- and KO-AMCM the basal RhoA activity was increased indicating that the tight regulation of RhoA in its signal context is disturbed when i) the sarcolemma is disorganized as after TAC, ii) cholesterol is depleted after MβCD treatment, or iii) p63RhoGEF is fully absent. This prompted the idea that p63RhoGEF is not only involved in the activation of RhoA in response to GPCRs, but also important for the membrane homeostasis in AMCM. By overexpression of p63RhoGEF, we could confirm its formerly described localization at the sarcolemma and found in addition a perinuclear distribution. This led amongst others to an increase in the number of cis-Golgi apparatus particles similar as observed in TAC-AMCM. In contrast, the loss of p63RhoGEF reduced particle number arguing that p63RhoGEF is involved in Golgi apparatus regulation. Interestingly, p63RhoGEF also affected the size of the AMCM nuclei. Overexpression of a dominant negative p63RhoGEF construct (p63ΔN) reduced the area of the nuclei, comparable as seen in isolated AMCM and intact myocardium after p63RhoGEF depletion. Opposing to this, in TAC-AMCM the mean nucleus area was increased. This was in accordance with the observed changes in cell size. Although there were differences in the type and height of the affected parameters, p63RhoGEF-overexpression and TAC increased cell size, whereas p63RhoGEF depletion reduced it. Summary: The presented data show for the first time that RhoA is activated in healthy adult cardiomyocytes in response to important cardiovascular GPCR ligands. Disturbances of the sarcolemmal organization ultimately lead to uncoupling of RhoA from its physiological signaling context. p63RhoGEF, verified as a direct mediator of the Gq/11 protein-dependent RhoA activation, plays an additional role in the regulation of RhoA and of membranous compartments in adult cardiomyocytes. Most importantly, its depletion leads to an uncoupling of RhoA from its signaling context." @default.
• W2948469013 created "2019-06-14" @default.
• W2948469013 creator A5029280685 @default.
• W2948469013 date "2022-02-21" @default.
• W2948469013 modified "2023-10-18" @default.
• W2948469013 title "Analysis of the GPCR-induced RhoA signaling in healthy and diseased adult cardiomyocytes" @default.
• W2948469013 cites W1236053900 @default.
• W2948469013 cites W1498975317 @default.
• W2948469013 cites W1500337621 @default.
• W2948469013 cites W1564750685 @default.
• W2948469013 cites W1568955905 @default.
• W2948469013 cites W1578175998 @default.
• W2948469013 cites W1583486328 @default.
• W2948469013 cites W1623913395 @default.
• W2948469013 cites W1730284679 @default.
• W2948469013 cites W1763096406 @default.
• W2948469013 cites W1764574939 @default.
• W2948469013 cites W1897848314 @default.
• W2948469013 cites W1901823973 @default.
• W2948469013 cites W1915765253 @default.
• W2948469013 cites W192394687 @default.
• W2948469013 cites W1933877372 @default.
• W2948469013 cites W1946469214 @default.
• W2948469013 cites W1963623014 @default.
• W2948469013 cites W1964236959 @default.
• W2948469013 cites W1965449074 @default.
• W2948469013 cites W1967135700 @default.
• W2948469013 cites W1967340614 @default.
• W2948469013 cites W1969566303 @default.
• W2948469013 cites W1970019262 @default.
• W2948469013 cites W1971687277 @default.
• W2948469013 cites W1972795887 @default.
• W2948469013 cites W1974279158 @default.
• W2948469013 cites W1976429225 @default.
• W2948469013 cites W1976843785 @default.
• W2948469013 cites W1977598234 @default.
• W2948469013 cites W1978018445 @default.
• W2948469013 cites W1978821056 @default.
• W2948469013 cites W1980549233 @default.
• W2948469013 cites W1983913693 @default.
• W2948469013 cites W1986587815 @default.
• W2948469013 cites W1987383823 @default.
• W2948469013 cites W1988097700 @default.
• W2948469013 cites W1989113874 @default.
• W2948469013 cites W1989434611 @default.
• W2948469013 cites W1990337343 @default.
• W2948469013 cites W1990392771 @default.
• W2948469013 cites W1991092434 @default.
• W2948469013 cites W1991426892 @default.
• W2948469013 cites W1992035790 @default.
• W2948469013 cites W1993722677 @default.
• W2948469013 cites W1994139415 @default.
• W2948469013 cites W1999538252 @default.
• W2948469013 cites W2000504183 @default.
• W2948469013 cites W2001442551 @default.
• W2948469013 cites W2003046582 @default.
• W2948469013 cites W2006828368 @default.
• W2948469013 cites W2007168414 @default.
• W2948469013 cites W2007366243 @default.
• W2948469013 cites W2007660746 @default.
• W2948469013 cites W2011751979 @default.
• W2948469013 cites W2019742616 @default.
• W2948469013 cites W2021075508 @default.
• W2948469013 cites W2021693681 @default.
• W2948469013 cites W2023758830 @default.
• W2948469013 cites W2023918318 @default.
• W2948469013 cites W2024732013 @default.
• W2948469013 cites W2025289160 @default.
• W2948469013 cites W2026339935 @default.
• W2948469013 cites W2026862467 @default.
• W2948469013 cites W2027501143 @default.
• W2948469013 cites W2030192558 @default.
• W2948469013 cites W2030485468 @default.
• W2948469013 cites W2030727312 @default.
• W2948469013 cites W2032295238 @default.
• W2948469013 cites W2033565692 @default.
• W2948469013 cites W2033841689 @default.
• W2948469013 cites W2034544118 @default.
• W2948469013 cites W2034941411 @default.
• W2948469013 cites W2036127937 @default.
• W2948469013 cites W2039919165 @default.
• W2948469013 cites W2042601656 @default.
• W2948469013 cites W2046610430 @default.
• W2948469013 cites W2047253447 @default.
• W2948469013 cites W2048777704 @default.
• W2948469013 cites W2050824263 @default.
• W2948469013 cites W2051673088 @default.
• W2948469013 cites W2051820912 @default.
• W2948469013 cites W2052780003 @default.
• W2948469013 cites W2052796543 @default.
• W2948469013 cites W2052926928 @default.
• W2948469013 cites W2056259311 @default.
• W2948469013 cites W2056801995 @default.
• W2948469013 cites W2056982707 @default.
• W2948469013 cites W2057937236 @default.
• W2948469013 cites W2058763646 @default.
• W2948469013 cites W2060071257 @default.
• W2948469013 cites W2060422904 @default.
• W2948469013 cites W2061508222 @default.
• W2948469013 cites W2065972506 @default.

• next