Cíl studie: Znalost mobility uretry patří mezi základní parametry urogynekologického vyšetření u žen se stresovým typem inkontinence. Z předchozích studií víme, že zvýšená mobilita uretry zvyšuje úspěšnost následné léčby stresového typu inkontinence moči. Hraniční hodnoty pro hypermobilitu jsou stanoveny jen arbitrárně, a to pomocí rotace úhlu gama o více než 30 resp. 50 stupňů či poklesu uretry o 15 mm při Valsalvově manévru. V rutinní klinické praxi kategorizujeme mobilitu při vyšetření uretry ultrazvukem buď jako nízkou, střední, nebo vysokou. Jsme však dostatečně přesní při subjektivním odhadu? K ověření přesnosti subjektivního odhadu jsme provedli retrospektivní analýzu ultrazvukového introitálního vyšetření mobility uretry a porovnali toto hodnocení s naměřenými parametry popisujícími mobilitu uretry, a to úhlu gama a vzdálenosti H. Typ studie: Retrospektivní studie. Název a sídlo pracoviště: Gynekologicko-porodnická klinika 1. LF UK a VFN, Praha. Metodika: Provedli jsme retrospektivní analýzu mobility uretry u žen s urodynamickou stresovou inkontinencí (USI), které byly následně operovány pro tuto diagnózu suburetrální páskou. U každé pacientky bylo před operací provedeno při ultrazvukovém vyšetření subjektivní semikvantitativní hodnocení mobility uretry. Subjektivní hodnocení bylo porovnáno s měřením, které bylo provedeno ex post z uložených volumů 4D ultrazvukového vyšetření. Měřili jsme vertikální vzdálenost uretrovezikální junkce (UVJ) od horizontály vycházející z dolního okraje symfýzy (vzdálenost H) v klidové pozici a při Valsalvově manévru. Mobilita byla počítána jako rozdíl obou naměřených hodnot. Stejným způsobem se počítala mobilita při použití druhého parametru – úhlu gama. Úhel gama svírá osa symfýzy a přímka spojující dolní okraj symfýzy a uretrovezikální junkci. Následně jsme porovnali subjektivní hodnocení a objektivně naměřené hodnoty. Dále jsme provedli vzájemnou korelaci úhlu gama a vzdálenosti H. Výsledky: V období 1/2009 až 10/2016 bylo operováno suburetrální páskou 427 žen s diagnózou USI. U celkem 393 byla dostupná všechna data. Průměrný věk ve skupině byl 56,5 let (min. 29 – max. 87, SD 7,9), průměrný BMI 27,4 (min. 18,3 – max. 39,6, SD 7,9) a průměrná parita 2,14. V celé skupině žen byla průměrná mobilita UVJ podle parametru H-vzdálenosti – 11,8 mm (min. -1; max. 37; SD 6,9) a průměrná mobilita, resp. rotace podle úhlu gama – 38,8 stupňů (min. -5; max. 118, SD 20,0). Při rozdělení do skupin podle subjektivního hodnocení mobility (nízká-, střední-, hypermobilita) byl podle obou objektivních parametrů pro hodnocení mobility rozdíl mezi skupinami signifikantní. Jako subjektivně nízká byla hodnocena průměrná mobilita 23,9 ° resp. 7,3 mm, střední 34,8 ° resp. 10,4 mm, a jako vysoká 48,6 ° resp. 15 mm. Při porovnání korelace obou objektivních parametrů jsme zjistili dobrou korelaci mezi hodnocením pomocí vzdálenosti H a úhlu gama, r = 0,693. Závěr: Subjektivní semikvantitativní hodnocení mobility uretry se ukazuje poměrně přesné pro rozdělení žen s tzv. nízkou, či naopak s hypermobilní uretrou.

Objective: The knowledge of the mobility of urethra plays important role in patients with stress urinary incontinence and its assessment is a part of standard urogynecological examination. It has been assumed that increased mobility is associated with higher likelihood of successful treatment. There is arbitrary defined cut-off for hypermobile urethra - descend of more than 15 mm or 30-degree resp. 50-degree rotation or more during Valsalva manoeuvre. Clinically we routinely categorize mobility of the urethra as high mobile or hypermobile urethras, low mobile urethra and the situation in-between as mobile urethras. But how accurate are we with such a subjective assessment? We have provided retrospective analysis of mobility of the urethra assessed during the clinical examination by transperineal ultrasound (US) with subjective scoring of the mobility (low, norm, hyper) and compared this assessment with detailed measurement of descent and rotation of the urethra. Design: Retrospective cohort study. Setting: Ob/Gyn department First Faculty of Medicine, Charles University and General University Hospital, Prague. Methods: This is a retrospective analysis of urethral mobility of women diagnosed with urodynamic stress incontinence (USI) and treated with tension free vaginal slings during the period 01/2009 – 10/2016. For each patient, there was available description of mobility at the time of preoperative assessment (low-, norm- or hyper-mobile) and we compared this assessment with measured parameters of bladder neck mobility analysed later from stored 4D US volumes. We have measured dorsocaudal movement of the bladder neck (BN) (H-distance). This is the distance of the BN from the horizontal line at the level of lower margin of the symphysis and we compared the position at rest and at Valsalva. Secondly, we measured rotation of the urethra using the gama angle - angle between the line connecting BN to lower margin of symphysis and axis of symphysis at rest and during the Valsalva manoeuvre. Mobility of the BN is the difference between the rest and Valsalva position. We compared the objective parameters of mobility with subjective assessment. We have provided correlation of both objective parameters. Results: 427 patients were treated during the analysed period, 393 had available stored 4D US volume for analysis. Mean age 56.5 years (min 29, max 87, SD 7.9), mean BMI 27.4 (min 18.3, max 39.6, SD 7.9), mean parity 2.14. Mean descent of the bladder neck was 11.8 mm (min -1, max 37; SD 6.9) Mean rotation of the bladder neck was 38.8 degree (min -5, max 118, SD 20.0). When we subjectively assessed the mobility as low the mean mobility was 23,9 ° - resp. 7.3 mm, for normal 34.8 ° resp. 10.4 mm and 48.6 ° resp. 15 mm for hypermobile uretras. We have found good correlation of both parameters - H-distance, gama angle - r = 0.693. Conclusion: Subjective assessment of mobility of the urethra seems to be reasonably accurate for distinguishing between low and hypermobile uretra.

• Klíčová slova
• mobilita uretrovezikální junkce, uretrovezikální junkce,
• MeSH
• diagnostické techniky urologické MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• předoperační vyšetření MeSH
• retrospektivní studie MeSH
• senioři MeSH
• stresová inkontinence moči * diagnóza   chirurgie   MeSH
• ultrasonografie metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• personální management MeSH
• profesní mobilita MeSH
• veřejné zdravotnictví MeSH
• zdravotní politika MeSH
• zdravotnický personál MeSH
• Geografické názvy
• Evropa MeSH

• Konspekt
• Veřejné zdraví a hygiena
• NLK Obory
• veřejné zdravotnictví
• NLK Publikační typ
• publikace WHO

PURPOSE OF STUDY Based on the selected criteria the study presents the evaluation of data of patients who underwent a surgery for proximal femoral fracture at the Department of Orthopaedic Trauma of the 1 st Faculty of Medicine and Military University Hospital Prague in the period 2013-2016. The paper aimed to determine whether there are any differences in proximal femoral fractures depending on the selected criteria. MATERIAL AND METHODS The study comprised 805 patients (562 women and 243 men) older than 21 years with proximal femoral fractures who were treated between January 1, 2013 and December 31, 2016. In the study, the patient s data on gender, age, a type of fracture (according to the AO classification), a level of mobility, time of injury and month of injury were analysed. The data was evaluated using the methods of descriptive statistics, one-way analysis of variance, two-way analysis by ANOVA and by following select POST-HOC test (Fisher LSD test) were utilized, 95% confidence interval was selected . RESULTS In the group of treated patients, the men were outnumbered by women (70% vs. 30%). In women the fractures occurred at a significantly (p < 0.001) older age (82 years) compared to men (77 years). Until the 8 th decade the fractures in men dominated, while from the 9th decade the fractures were more frequent in women. A sharp increase in the number of fractures in women was reported exactly in the 9th decade (56% of the total number of fractures in women). The group of women showed a significant difference (p = 0.027) between the age of femoral neck fractures and the age of trochanteric femoral fractures (80 years vs. 84 years). In both genders intracapsular and pertrochanteric fractures prevailed. In women and men with a limited mobility, fractures were sustained at the highest mean age (85 years in women, 84 years in men) compared to the other levels of mobility. In mobile patients, a significant difference (p < 0.001) was found between the age of women and men at which the fracture was sustained (79 years vs. 71 years). The largest significant difference (p < 0.001) in the age of fracture is seen in night time (84 years in women, 71 years in men). The highest number of injuries was suffered in the months of October to January, whereas the lowest number was reported in the period from June to July. DISCUSSION The epidemiological studies underline a notably higher incidence of proximal femoral fractures in women than in men, which was also confirmed by our study. The mean age of women with proximal femoral fracture was significantly higher compared to men. Similar conclusions have been drawn also by other studies. The incidence of fractures in dependence on the age group was the highest in women and men in their 9th decade, which compared to the previous studies was at a later age. This finding can be justified by aging of the Czech population, with the growing number of persons in the 9 th decade age group between 2012 and 2017 by 40,000. Nonetheless, men prevailed in fractures until the 9 th decade, while from the 9 th decade there is a marked growth in the number of these fractures sustained by women. This corresponds with the findings of other authors. As to the individual types of fractures (neck, trochanteric fracture) no difference was revealed in the percentage of women and men. In both genders intracapsular and petrochanteric fractures prevailed. Neither the level of pre-fracture mobility, nor the time of injury have been sufficiently monitored and evaluated with regard to the age of patients, therefore there is no adequate data to compare our results with. The lowest mean age at the time of injury was achieved by fully mobile men (71 years), which can be attributable to their sports and social activities performed at this age. The highest number of proximal femoral fractures was reported in the period from October to January, whereas their lowest number was reported in the period from June to July, which corresponds with the majority of other conducted studies in this area of research. CONCLUSIONS A considerably higher incidence of proximal femoral fractures in women was confirmed. The mean age of women with proximal femoral fracture was significantly higher than in men (82 vs. 77 years). Men prevailed in the number of fractures up to the 9 th decade, whereas from the 9 th decade the number of proximal femoral fractures in women significantly grew. In individual types of fractures (neck, trochanteric fractures), no difference in percentage of women and men was found. In both the genders intracapsular and pertrochanteric fractures significantly prevailed. The lowest mean age (71 years) at the time of injury was achieved by fully mobile men. The same age was identified in men also with respect to injuries sustained at night. The highest number of proximal femoral fractures was reported in the period from October to January, the lowest in the period from June to July. The knowledge of these seasonal variations can help plan the health care in the medical facility concerned. Key words: proximal femur, fracture, gender, mobility, day-time, seasonality.

• MeSH
• dospělí MeSH
• fraktury femuru * epidemiologie   chirurgie   MeSH
• fraktury krčku femuru * epidemiologie   chirurgie   MeSH
• fraktury kyčle * epidemiologie   chirurgie   MeSH
• incidence MeSH
• lidé středního věku MeSH
• lidé MeSH
• omezení pohyblivosti MeSH
• rizikové faktory MeSH
• roční období MeSH
• senioři MeSH
• sexuální faktory MeSH
• věkové faktory MeSH
• vnitřní fixace fraktury * metody   statistika a číselné údaje   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Amyotrofická laterálna skleróza (ALS) je pomerne zriedkavé neuromuskulárne ochorenie charakterizované degeneráciou dolného a horného motorického neurónu. ALS zahŕňa širokú škálu symptómov, vrátane svalovej slabosti, kŕčov, únavy, spasticity, dysfágie, dysartrie, respiračného zlyhania, a tiež kognitívne zmeny a zmeny nálady. Rehabilitácia tvorí dôležitú časť terapie. Hlavným cieľom rehabilitácie je zabránenie atrofie svalov a zvýšenie svalovej sily. Zachovanie dostatočnej mobility pomocou cvičení tvorí ďalšiu podstatnú časť pohybovej liečby. Cvičenia zamerané na zvýšenie rozsahu pohyblivosti tvoria dôležitú časť liečby v prevencii a liečbe kontraktúr. Potenciálnu úlohu plní tiež v zlepšení spasticity. Pleiotropný efekt cvičenia sa uplatňuje aj pri zlepšení kvality života pacientov. Benefitom pohybovej liečby je aj ovplyvnenie únavy. Respiračná rehabilitácia môže pomôcť predchádzať respiračným komplikáciám. Celkovo, rehabilitačná liečba môže napomôcť v zlepšení funkčného statusu pacientov, svalovej sily, a tiež zlepšiť kvalitu života pacientov.

Amyotrophic lateral sclerosis (ALS) is a relatively rare neuromuscular disease characterized by degeneration of the lower and upper motor neurons. ALS includes a wide range of symptoms, including muscle weakness, cramps, fatigue, spasticity, dysphagia, dysarthria, respiratory failure, and also cognitive and mood changes. Rehabilitation is an important part of therapy. The main goal of rehabilitation is to prevent muscle atrophy and increase muscle strength. Maintaining adequate mobility through exercise is another essential part of exercise therapy. Exercises aimed at increasing the range of motion are an important part of treatment in the prevention and treatment of contractures. They also play a potential role in improving spasticity. Quality of patients' life is expected to improve also due to pleiotropic effect of exercise. Exercise therapy may also improve fatigue. Respiratory rehabilitation can help prevent respiratory complications. Overall, rehabilitation therapy can help improve patients functional status, muscle strength and patients quality of life as well.

• MeSH
• amyotrofická laterální skleróza * rehabilitace   MeSH
• cvičení MeSH
• lidé MeSH
• neurorehabilitace metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

There are some indications that biased μ-opioid ligands may diversely affect μ-opioid receptor (MOR) properties. Here, we used confocal fluorescence recovery after photobleaching (FRAP) to study the regulation by different MOR agonists of receptor movement within the plasma membrane of HEK293 cells stably expressing a functional yellow fluorescent protein (YFP)-tagged μ-opioid receptor (MOR-YFP). We found that the lateral mobility of MOR-YFP was increased by (D-Ala(2),N-MePhe(4),Gly(5)-ol)-enkephalin (DAMGO) and to a lesser extent also by morphine but decreased by endomorphin-2. Interestingly, cholesterol depletion strongly enhanced the ability of morphine to elevate receptor mobility but significantly reduced or even eliminated the effect of DAMGO and endomorphin-2, respectively. Moreover, the ability of DAMGO and endomorphin-2 to influence MOR-YFP movement was diminished by pertussis toxin treatment. The results obtained by agonist-stimulated [(35)S]GTPγS binding assays indicated that DAMGO exhibited higher efficacy than morphine and endomorphin-2 did and that the efficacy of DAMGO, contrary to the latter agonists, was enhanced by cholesterol depletion. Overall, our study provides clear evidence that biased MOR agonists diversely affect receptor mobility in plasma membranes as well as MOR/G protein coupling and that the regulatory effect of different ligands depends on the membrane cholesterol content. These findings help to delineate the fundamental properties of MOR regarding their interaction with biased MOR ligands and cognate G proteins.

• MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• cholesterol nedostatek   MeSH
• enkefalin, Ala(2)-MePhe(4)-Gly(5)- metabolismus   farmakologie   MeSH
• FRAP MeSH
• guanosin 5'-O-(3-thiotrifosfát) metabolismus   MeSH
• HEK293 buňky MeSH
• konfokální mikroskopie MeSH
• lidé MeSH
• ligandy MeSH
• luminescentní proteiny genetika   metabolismus   MeSH
• morfin metabolismus   farmakologie   MeSH
• narkotika - antagonisté farmakologie   MeSH
• oligopeptidy metabolismus   farmakologie   MeSH
• pertusový toxin farmakologie   MeSH
• proteiny vázající GTP - alfa-podjednotky Gi-Go metabolismus   MeSH
• receptory opiátové mu agonisté   genetika   metabolismus   MeSH
• rekombinantní fúzní proteiny metabolismus   MeSH
• transfekce MeSH
• transport proteinů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Increased sedentism during the Holocene has been proposed as a major cause of decreased skeletal robusticity (bone strength relative to body size) in modern humans. When and why declining mobility occurred has profound implications for reconstructing past population history and health, but it has proven difficult to characterize archaeologically. In this study we evaluate temporal trends in relative strength of the upper and lower limb bones in a sample of 1,842 individuals from across Europe extending from the Upper Paleolithic [11,000-33,000 calibrated years (Cal y) B.P.] through the 20th century. A large decline in anteroposterior bending strength of the femur and tibia occurs beginning in the Neolithic (∼ 4,000-7,000 Cal y B.P.) and continues through the Iron/Roman period (∼ 2,000 Cal y B.P.), with no subsequent directional change. Declines in mediolateral bending strength of the lower limb bones and strength of the humerus are much smaller and less consistent. Together these results strongly implicate declining mobility as the specific behavioral factor underlying these changes. Mobility levels first declined at the onset of food production, but the transition to a more sedentary lifestyle was gradual, extending through later agricultural intensification. This finding only partially supports models that tie increased sedentism to a relatively abrupt Neolithic Demographic Transition in Europe. The lack of subsequent change in relative bone strength indicates that increasing mechanization and urbanization had only relatively small effects on skeletal robusticity, suggesting that moderate changes in activity level are not sufficient stimuli for bone deposition or resorption.

• MeSH
• biomechanika MeSH
• dějiny starověku MeSH
• lidé MeSH
• sedavý životní styl * MeSH
• zásobování potravinami * MeSH
• zemědělství MeSH
• zkameněliny MeSH
• Check Tag
• dějiny starověku MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• historické články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Geografické názvy
• Evropa MeSH

Kontinuálna laterálna rotačná terapia, dnes už rozšírená v západných krajinách sveta, je overenou prídavnou intervenciou u pacientov hospitalizovaných na pracoviskách intenzívnej starostlivosti. Je využívaná u intubovaných alebo tracheostomovaných pacientov na umelej ventilácii pľúc, v prevencii komplikácií z imobility. Najvýznamnejšiu úlohu má pri prevencii ventilátorových pneumónií, ako aj iných pľúcnych komplikácií umelej ventilácie pľúc, či ich liečbe. Ďalšou oblasťou jej využitia môže byť napríklad včasná mobilizácia pacienta, či eventuálne aj prevencia dekubitov na cielených predilekčných miestach u imobilných pacientov. Lôžka s funkciou laterálneho náklonu sa však postupne stávajú súčasťou aj našich pracovísk a teda aj zručností sestier, pracujúcich v intenzívnej starostlivosti. Preto je potrebné vzdelávať sa v oblasti správneho výberu vhodných pacientov pre túto formu terapie, jej zásadami a obmedzeniami, ako aj rizikami, či kontraindikáciamiContinuous lateral rotational therapy

Continuous lateral rotational therapy nowadays widely spread in western countries of the world is one verified additional intervention in patients hospitalized at the intensive care departments. It is used in intubated patients or patient with tracheostomy on mechanical ventilation, in the prevention of immobility complications. The most important role plays the lateral rotational therapy in prevention of ventilator associated pneumonia and other lung complications of mechanical ventilation or their treatment. Next area of its use can be for example early mobilization of patient or eventually decubitus prevention in aimed predilection places in immobile patients. Beds with the lateral tilting function are progressively becoming a part of our workplaces also the part of the skills of nurses working in intensive care. Therefore, it is to educate in the area of right selection of the appropriate patient for this form therapy, its principles and limitations, also the risks or contraindications needed.

• Klíčová slova
• laterální náklon,
• MeSH
• dekubity prevence a kontrola   MeSH
• lůžka trendy   MeSH
• muskuloskeletální manipulace metody   přístrojové vybavení   MeSH
• péče o pacienty v kritickém stavu MeSH
• polohování pacienta MeSH
• ventilátorová pneumonie prevence a kontrola   MeSH

The dual-affinity nitrate transceptor NITRATE TRANSPORTER1.1 (NRT1.1) has two modes of transport and signaling, governed by Thr-101 (T101) phosphorylation. NRT1.1 regulates lateral root (LR) development by modulating nitrate-dependent basipetal auxin export and nitrate-mediated signal transduction. Here, using the Arabidopsis (Arabidopsis thaliana) NRT1.1T101D phosphomimetic and NRT1.1T101A nonphosphorylatable mutants, we found that the phosphorylation state of NRT1.1 plays a key role in NRT1.1 function during LR development. Single-particle tracking revealed that phosphorylation affected NRT1.1 spatiotemporal dynamics. The phosphomimetic NRT1.1T101D form showed fast lateral mobility and membrane partitioning that facilitated auxin flux under low-nitrate conditions. By contrast, nonphosphorylatable NRT1.1T101A showed low lateral mobility and oligomerized at the plasma membrane (PM), where it induced endocytosis via the clathrin-mediated endocytosis and microdomain-mediated endocytosis pathways under high-nitrate conditions. These behaviors promoted LR development by suppressing NRT1.1-controlled auxin transport on the PM and stimulating Ca2+-ARABIDOPSIS NITRATE REGULATED1 signaling from the endosome.

• MeSH
• Arabidopsis genetika   růst a vývoj   metabolismus   MeSH
• dusičnany metabolismus   MeSH
• fosforylace MeSH
• kořeny rostlin růst a vývoj   MeSH
• kyseliny indoloctové metabolismus   MeSH
• proteiny huseníčku metabolismus   MeSH
• proteiny přenášející anionty genetika   metabolismus   MeSH
• rostlinné proteiny genetika   metabolismus   MeSH
• transkripční faktory metabolismus   MeSH
• vápníková signalizace MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

β-Arrestins are known to play a crucial role in GPCR-mediated transmembrane signaling processes. However, there are still many unanswered questions, especially those concerning the presumed similarities and differences of β-arrestin isoforms. Here, we examined the roles of β-arrestin 1 and β-arrestin 2 at different levels of μ-opioid receptor (MOR)-regulated signaling, including MOR mobility, internalization of MORs, and adenylyl cyclase (AC) activity. For this purpose, naïve HEK293 cells or HEK293 cells stably expressing YFP-tagged MOR were transfected with appropriate siRNAs to block in a specific way the expression of β-arrestin 1 or β-arrestin 2. We did not find any significant differences in the ability of β-arrestin isoforms to influence the lateral mobility of MORs in the plasma membrane. Using FRAP and line-scan FCS, we observed that knockdown of both β-arrestins similarly increased MOR lateral mobility and diminished the ability of DAMGO and endomorphin-2, respectively, to enhance and slow down receptor diffusion kinetics. However, β-arrestin 1 and β-arrestin 2 diversely affected the process of agonist-induced MOR endocytosis and exhibited distinct modulatory effects on AC function. Knockdown of β-arrestin 1, in contrast to β-arrestin 2, more effectively suppressed forskolin-stimulated AC activity and prevented the ability of activated-MORs to inhibit the enzyme activity. Moreover, we have demonstrated for the first time that β-arrestin 1, and partially β-arrestin 2, may somehow interact with AC and that this interaction is strongly supported by the enzyme activation. These data provide new insights into the functioning of β-arrestin isoforms and their distinct roles in GPCR-mediated signaling.

• MeSH
• adenylátcyklasy * metabolismus   MeSH
• beta arrestin 1 metabolismus   MeSH
• beta arrestin 2 metabolismus   MeSH
• beta arrestiny metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• receptory opiátové mu * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The WNT receptors of the Frizzled family comprise ten mammalian isoforms, bind WNT proteins and mediate downstream signaling to regulate stem cell fate, neuronal differentiation, cell survival and more. WNT-induced signaling pathways are either β-catenin-dependent or -independent, thereby dividing the 19 mammalian WNT proteins into two groups. So far hardly any quantitative, pharmacological information is available about WNT-FZD interaction profiles, affinities or mechanisms of signaling specification through distinct WNT/FZD pairings. This lack of knowledge originates from difficulties with WNT purification and a lack of suitable assays, such as ligand binding assays and FZD activity readouts. In order to minimize this gap, we employ fluorescence recovery after photobleaching (FRAP) to investigate WNT effects on the lateral mobility of FZD6-GFP in living cells. Pharmacological uncoupling of heterotrimeric G proteins by pertussis toxin and N-ethylmaleimide argues that changes in FZD6 mobility are related to putative precoupling of heterotrimeric Gi/o proteins to FZD6. We show that recombinant WNT-1, -2, 3A, -4, -5A, -7A, -9B and -10B affect FZD6 surface mobility and thus act on this receptor. WNT-5B and WNT-11, on the other hand, have no effect on FZD6 mobility and we conclude that they do not act through FZD6. We introduce here a novel way to assess WNT-FZD interaction by live cell imaging allowing further mapping of WNT-FZD interactions and challenging previous experimental limitations. Increased understanding of WNT-FZD selectivity provides important insight into the biological function of this crucial signaling system with importance in developmental biology, stem cell regulation oncogenesis, and human disease.

• MeSH
• buněčná membrána metabolismus   MeSH
• ethylmaleimid farmakologie   MeSH
• FRAP * MeSH
• frizzled receptory agonisté   genetika   metabolismus   MeSH
• HEK293 buňky MeSH
• heterotrimerní G-proteiny metabolismus   MeSH
• lidé MeSH
• pertusový toxin toxicita   MeSH
• proteiny vázající GTP - alfa-podjednotky Gi-Go metabolismus   MeSH
• proteiny Wnt genetika   metabolismus   MeSH
• protoonkogenní proteiny genetika   metabolismus   MeSH
• rekombinantní proteiny genetika   metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH