Cell polarity is a key feature in the development of multicellular organisms. For instance, asymmetrically localized plasma-membrane-integral PIN-FORMED (PIN) proteins direct transcellular fluxes of the phytohormone auxin that govern plant development. Fine-tuned auxin flux is important for root protophloem sieve element differentiation and requires the interacting plasma-membrane-associated BREVIS RADIX (BRX) and PROTEIN KINASE ASSOCIATED WITH BRX (PAX) proteins. We observed "donut-like" polar PIN localization in developing sieve elements that depends on complementary, "muffin-like" polar localization of BRX and PAX. Plasma membrane association and polarity of PAX, and indirectly BRX, largely depends on phosphatidylinositol-4,5-bisphosphate. Consistently, mutants in phosphatidylinositol-4-phosphate 5-kinases (PIP5Ks) display protophloem differentiation defects similar to brx mutants. The same PIP5Ks are in complex with BRX and display "muffin-like" polar localization. Our data suggest that the BRX-PAX module recruits PIP5Ks to reinforce PAX polarity and thereby the polarity of all three proteins, which is required to maintain a local PIN minimum.
- MeSH
- Arabidopsis genetika růst a vývoj metabolismus MeSH
- buněčná diferenciace * MeSH
- buněčná membrána metabolismus MeSH
- fosfatasy genetika metabolismus MeSH
- fosfotransferasy s alkoholovou skupinou jako akceptorem metabolismus MeSH
- kořeny rostlin genetika růst a vývoj metabolismus MeSH
- mutace MeSH
- polarita buněk * MeSH
- proteiny huseníčku genetika metabolismus MeSH
- regulace genové exprese u rostlin * MeSH
- transkripční faktory paired box genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Biphenotypic sinonasal sarcoma (BSNS) is a recently recognized type of sarcoma arising exclusively in the sinonasal tract displaying unique clinical course, histopathology, and genetics. Due to its rarity, only case series and case reports are available. In order to provide an overview of the current understanding of this disease, we present a comprehensive review of the literature and present three previously unreported cases of BSNS. A total of 55 genetically characterized and 41 cases without molecular data were identified in the literature. Two-thirds of patients were female and the peak incidence was in the fifth decade. Fatal outcome was rare (two cases with intracranial extension) and local recurrence occurred in 31.6%, all occurring within 5 years after initial treatment. Histologically, BSNS is highly cellular in the majority of cases and composed of fascicles of spindle cells, with entrapped hyperplastic surface epithelium being a frequent finding. The immunohistochemical profile is characteristic due to the biphasic nature of this lesion, with shared features of both myogenic and neural origin. Rhabdomyoblastic differentiation is apparent in a subset of cases. The most common genetic event is the PAX3-MAML3 fusion (58.6%) but isolated PAX3 rearrangement (19.2%), absence of rearrangements (9.1%), PAX3-FOXO1 (8.1%), PAX3-NCOA1 (4%), and isolated MAML3 rearrangement (2%) have also been reported. In conclusion, the recognition of BSNS is crucial due to its relatively indolent clinical course. A selected immunohistochemical panel and/or molecular confirmation can be used to aid in appropriate diagnosis and consequently in prognostication and to avoid overtreatment with chemotherapy regimens used in its mimics.
- MeSH
- DNA vazebné proteiny genetika MeSH
- dospělí MeSH
- fenotyp MeSH
- forkhead box protein O1 genetika MeSH
- fúze genů MeSH
- fúzní onkogenní proteiny genetika MeSH
- genová přestavba MeSH
- imunohistochemie MeSH
- jaderné proteiny genetika MeSH
- koaktivátor 1 jaderných receptorů genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové biomarkery genetika MeSH
- nádory vedlejších dutin nosních diagnóza genetika patologie MeSH
- prognóza MeSH
- sarkom diagnóza genetika patologie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- transkripční faktor PAX3 genetika MeSH
- transkripční faktory paired box genetika MeSH
- transkripční faktory genetika MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- přehledy MeSH
Wnt/β-catenin signaling plays an essential role in the retinal pigment epithelium (RPE) determination. Since activity of Pax6 (together with Pax2) is also required for the RPE determination, we investigated a possible genetic interaction between Pax6 and Wnt/β-catenin signaling pathway by analyzing Pax6, β-catenin, and Pax6/β-catenin conditional knockout mice. Although Pax6 inactivation alone had no impact on initial specification determined by the expression of Mitf and Otx2, melanin pigmentation was reduced in the RPE. This suggests that along with Mitf and Otx2, Pax6 is required for the full differentiation of RPE. Reporter gene assays in vitro suggest that hypopigmentation is at least in part due to the direct regulation of genes encoding enzymes involved in melanin synthesis by Pax6, Mitf, and β-catenin. The RPE of a β-catenin/Pax6 double mutant was differentiated into the neural retina; however, the tissue was thinner than that of the conditional β-catenin mutant due to reduced proliferation. Together, our data demonstrate that Pax6 is required for the RPE differentiation by regulating pigmentation and accountable for hyperproliferation in the transdifferentiated RPE. In this context, Pax6 appears to function as a pleiotropic regulator, directing development of ocular tissues in concert with the signaling pathway and, at the same time, regulating expression of structural component of the eye, such as shielding pigment.
- MeSH
- beta-katenin genetika metabolismus MeSH
- homeodoménové proteiny genetika metabolismus MeSH
- myši transgenní MeSH
- myši MeSH
- oční proteiny genetika metabolismus MeSH
- represorové proteiny genetika metabolismus MeSH
- retina cytologie metabolismus MeSH
- retinální pigmentový epitel metabolismus MeSH
- signální dráha Wnt * MeSH
- transdiferenciace buněk MeSH
- transkripční faktory paired box genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvod: vrozená izolovaná aniridie je vzácné postižení struktur obou očí projevující se především chyběním duhovky, fotofobií a sníženým viděním. V klinickém obrazu může být přítomný nystagmus, strabismus, ptóza víčka, amblyopie, vyšší refrakční vada, anisometropie, rohovkové změny, patologie čočky, dysgeneze struktur komorového úhlu, hypoplazie makuly a zrakového nervu, kongenitální nebo sekundární glaukom. Tento typ dysgeneze oka je nejčastěji podmíněn mutací PAX6. Cílem naší práce bylo popsat oftalmologické nálezy členů rodiny s výskytem dědičné aniridie (MIM #106210), diskutovat jejich závažnost, prognózu a možnosti terapie. Materiál a metodika: zhodnocení předchozích klinických záznamů a oftalomogické vyšetření proběhlo u 4 osob z tří generací rodiny s výskytem izolované vrozené aniridie. Dle možností a compliance byla u pacientů provedena tato vyšetření: vízus, nitrooční tenze, refrakce, vyšetření štěrbinovou lampou, biomikroskopické vyšetření, pachymetrie, OCT. Dále bylo provedeno klinicko genetické vyšetření a následně sekvenace PAX6. Výsledky: u všech vyšetřených osob byla zjištěna plně vyjádřená závažná symptomatologie, byla pozorována přítomnost aniridie a fotofobie. Pozitivní korelaci s věkem vykazoval progresivní pokles vizu k praktické slepotě v důsledku keratopatie asociované s aniridií, sekundárním glaukomem a kataraktou. DNA analýza u všech postižených osob odhalila přítomnost mutace p.Gln180X v PAX6. Tato mutace vede k tvorbě zkráceného atak nefunkčního PAX6 proteinu. Závěr: mutace v genu PAX6, které vedou k předčasné terminaci tvorby bílkoviny, jsou spojené často s těžkým průběhem onemocnění as malou variabilitou závažnosti postižení intrafamiliárně. Tento fakt dokumentuje irodina popsaná v této práci, kdy projevy upacientů jsou závažné aprogredují s věkem. Terapie je svízelná, často jen parciálně úspěšná, jako je tomu iv případě sekundárního glaukomu mladé pacientky. Každý operační zákrok je nutno individuálně vždy velmi pečlivě uvážit. Navíc utéto nemoci dochází kneúprosné progresi očního nálezu spostupným zhoršováním itak již většinou chabé zrakové ostrosti až do praktické slepoty. Klíčová slova: aniridia, PAX6, hypoplazie makuly, glaukom
Background: inborn isolated aniridia is rare bilateral impairment of several eye structures manifesting mainly by absence of iris, photophobia and decreased visual acuity. There are also others ocular symptoms associated with aniridia such as nystagmus, strabismus, eyelid ptosis, amblyopia, serious refractive errors, anisometropia, corneal changes, impairment of the lens, chamber angle dysgenesis, optic nerve and macular hypoplasia and congenital or secondary glaucoma. The most frequent aetiology of this eye dysgenesis is mutation in PAX6. Aim of this report is to describe ocular findings in the family with familial aniridia (MIM #106210), to debate their severity, prognosis and therapy options. Material and methods: assessment of previous medical history and actual ophthalmological findings in 4 persons of 3 generation family with aniridia. According to the compliance, the patients underwent these tests: assessment of the visual acuity, intraocular pressure, refraction test, slit-lamp examination and biomicroscopy, pachymetry test and OCT examination. The genetic counselling was performed with subsequent PAX6 mutation analysis. Results: all of the examined aniridia family members showed severe symptoms of the disease, the aniridia and photophobia were present. Positive age related correlation showed progressive visual acuity decrease to the practical blindness due to aniridia-associated keratopathy, secondary glaucoma and cataract. DNA analysis revealed presence of p.Gln180X PAX6 mutation in all of the affected persons. The mutation leads to shortened and therefore non-functional protein. Conclusions: PAX6 mutations leading to premature termination of protein translation are frequently associated with severe symptoms of aniridia and small intrafamilial variability of ocular impairment. This fact is also well demonstrated in members of family described by this report, the symptoms are severe and progressing with age. Therapy is difficult and often with partial success, such in case of secondary glaucoma in young girl from this family. Any eye surgery must be individually judged due to risk of several post-operative complications. And more, the poor vision in aniridia patients is progressively worsening in time to practical blindness. Key words: aniridia, PAX6, macular hypoplasia, glaucoma
- Klíčová slova
- hypoplazie makuly,
- MeSH
- aniridie * diagnóza genetika patofyziologie terapie MeSH
- diagnostické techniky oftalmologické MeSH
- dítě MeSH
- dospělí MeSH
- fotofobie etiologie MeSH
- glaukom etnologie farmakoterapie MeSH
- homeodoménové proteiny genetika MeSH
- katarakta etiologie MeSH
- lidé MeSH
- mutace MeSH
- nemoci rohovky etiologie MeSH
- oční proteiny * genetika MeSH
- patologický nystagmus etiologie MeSH
- progrese nemoci MeSH
- represorové proteiny genetika MeSH
- retina patologie MeSH
- rodokmen MeSH
- rohovka patologie MeSH
- sekvenční analýza DNA MeSH
- senioři MeSH
- strabismus etiologie MeSH
- transkripční faktory paired box genetika MeSH
- zraková ostrost MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
The physical contact of optic vesicle with head surface ectoderm is an initial event triggering eye morphogenesis. This interaction leads to lens specification followed by coordinated invagination of the lens placode and optic vesicle, resulting in formation of the lens, retina and retinal pigmented epithelium. Although the role of Pax6 in early lens development has been well documented, its role in optic vesicle neuroepithelium and early retinal progenitors is poorly understood. Here we show that conditional inactivation of Pax6 at distinct time points of mouse neuroretina development has a different impact on early eye morphogenesis. When Pax6 is eliminated in the retina at E10.5 using an mRx-Cre transgene, after a sufficient contact between the optic vesicle and surface ectoderm has occurred, the lens develops normally but the pool of retinal progenitor cells gradually fails to expand. Furthermore, a normal differentiation program is not initiated, leading to almost complete disappearance of the retina after birth. By contrast, when Pax6 was inactivated at the onset of contact between the optic vesicle and surface ectoderm in Pax6(Sey/flox) embryos, expression of lens-specific genes was not initiated and neither the lens nor the retina formed. Our data show that Pax6 in the optic vesicle is important not only for proper retina development, but also for lens formation in a non-cell-autonomous manner.
- MeSH
- buněčná diferenciace genetika fyziologie MeSH
- embryonální kmenové buňky cytologie metabolismus MeSH
- genový knockdown MeSH
- homeodoménové proteiny antagonisté a inhibitory genetika metabolismus MeSH
- kontrolní body buněčného cyklu genetika fyziologie MeSH
- myši knockoutované MeSH
- myši transgenní MeSH
- myši MeSH
- nervové kmenové buňky cytologie metabolismus MeSH
- oční čočka embryologie metabolismus MeSH
- oční proteiny antagonisté a inhibitory genetika metabolismus MeSH
- represorové proteiny antagonisté a inhibitory genetika metabolismus MeSH
- retina cytologie embryologie metabolismus MeSH
- těhotenství MeSH
- trans-aktivátory genetika metabolismus MeSH
- transkripční faktory paired box antagonisté a inhibitory genetika metabolismus MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- hypoplazie makuly,
- MeSH
- aniridie * diagnóza genetika patofyziologie terapie MeSH
- diagnostické techniky oftalmologické MeSH
- dítě MeSH
- dospělí MeSH
- fotofobie etiologie MeSH
- glaukom etnologie farmakoterapie MeSH
- homeodoménové proteiny genetika MeSH
- katarakta etiologie MeSH
- lidé MeSH
- mutace MeSH
- nemoci rohovky etiologie MeSH
- oční proteiny * genetika MeSH
- patologický nystagmus etiologie MeSH
- progrese nemoci MeSH
- represorové proteiny genetika MeSH
- retina patologie MeSH
- rodokmen MeSH
- rohovka patologie MeSH
- sekvenční analýza DNA MeSH
- senioři MeSH
- strabismus etiologie MeSH
- transkripční faktory paired box genetika MeSH
- zraková ostrost MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Tumor suppressor genes (TSGs) are commonly inactivated by somatic mutation and/or promoter methylation; yet, recent high-throughput genomic studies have not identified key TSGs inactivated by both mechanisms. We pursued an integrated molecular analysis based on methylation binding domain sequencing (MBD-seq), 450K Methylation arrays, whole exome sequencing, and whole genome gene expression arrays in primary head and neck squamous cell carcinoma (HNSCC) tumors and matched uvulopalatopharyngoplasty tissue samples (UPPPs). We uncovered 186 downregulated genes harboring cancer specific promoter methylation including PAX1 and PAX5 and we identified 10 key tumor suppressor genes (GABRB3, HOXC12, PARP15, SLCO4C1, CDKN2A, PAX1, PIK3AP1, HOXC6, PLCB1, and ZIC4) inactivated by both promoter methylation and/or somatic mutation. Among the novel tumor suppressor genes discovered with dual mechanisms of inactivation, we found a high frequency of genomic and epigenomic alterations in the PAX gene family of transcription factors, which selectively impact canonical NOTCH and TP53 pathways to determine cell fate, cell survival, and genome maintenance. Our results highlight the importance of assessing TSGs at the genomic and epigenomic level to identify key pathways in HNSCC, deregulated by simultaneous promoter methylation and somatic mutations.
- MeSH
- aktivátorový protein specifický pro B-buňky genetika metabolismus MeSH
- CpG ostrůvky MeSH
- kohortové studie MeSH
- lidé MeSH
- metylace DNA * MeSH
- mutace MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 genetika metabolismus MeSH
- nádory hlavy a krku genetika metabolismus MeSH
- promotorové oblasti (genetika) * MeSH
- receptor Notch1 genetika metabolismus MeSH
- spinocelulární karcinom genetika metabolismus MeSH
- transkripční faktory paired box genetika metabolismus MeSH
- tumor supresorové geny * MeSH
- umlčování genů * MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Lens formation in mouse is critically dependent on proper development of the retinal neuroectoderm that is located close beneath the head surface ectoderm. Signaling from the prospective retina triggers lens-specific gene expression in the surface-ectoderm. Supression of canonical Wnt/beta-catenin signaling in the surface ectoderm is one of the prerequisites for lens development because, as we show here, ectopic Wnt activation in the retina and lens abrogates lens formation. Wnt inhibiton is mediated by signals coming from the retina but its exact mechanism is unknown. We show that Pax6 directly controls expression of several Wnt inhibitors such as Sfrp1, Sfrp2, and Dkk1 in the presumptive lens. In accordance, absence of Pax6 function leads to aberrant canonical Wnt activity in the presumptive lens that subsequently impairs lens development. Thus Pax6 is required for down-regulation of canonical Wnt signaling in the presumptive lens ectoderm.
- MeSH
- beta-katenin genetika metabolismus MeSH
- ektoderm metabolismus MeSH
- embryo savčí metabolismus MeSH
- homeodoménové proteiny genetika metabolismus MeSH
- morfogeneze genetika MeSH
- myši transgenní MeSH
- myši MeSH
- oční čočka embryologie metabolismus MeSH
- oční proteiny genetika metabolismus MeSH
- proteiny Wnt metabolismus MeSH
- represorové proteiny genetika metabolismus MeSH
- retina metabolismus MeSH
- signální transdukce genetika MeSH
- transkripční faktory paired box genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of our study was to characterize mouse embryonal carcinoma (EC) cells P19 in different stages of retinoic acid induced neurodifferentiation by two methods, immunocytochemistry and RT qPCR. The characterization of the cells is crucial before any transplantation into any model, e.g. in our case into the mouse brain with the aim to treat a neurodegenerative disease. Specific protein markers (MAP-2, OCT-4, FORSE-1) were detected by immunocytochemistry in the cell cultures. The mRNA expression levels of PAX-6, MASH-1, Brachyury, GATA-4 and AFP were determined by RT qPCR method. HPRT was used as a housekeeping gene. The degree of differentiation can be characterized by expression of analyzed genes. The presence of OCT-4 and FORSE-1 proteins in undifferentiated pluripotent cells and the presence of dendrite specific MAP-2 in neuroprogenitors was detected. The expression levels of PAX-6 and MASH-1 increased and expression of Brachyury decreased during the neurodifferentiation process. The expression levels of GATA-4 and AFP were the highest after induction of differentiation with retinoic acid. Detailed characterization of cells before transplantation experiments can contribute to better understanding of their effect.
- MeSH
- antigeny povrchové genetika metabolismus MeSH
- buněčná diferenciace genetika účinky léků MeSH
- exprese genu MeSH
- financování organizované MeSH
- homeodoménové proteiny genetika metabolismus MeSH
- imunohistochemie MeSH
- kmenové buňky embryonálního karcinomu fyziologie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádorové buňky kultivované MeSH
- oční proteiny genetika metabolismus MeSH
- oktamerní transkripční faktor 3 genetika metabolismus MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proteiny asociované s mikrotubuly genetika metabolismus MeSH
- represorové proteiny genetika metabolismus MeSH
- transkripční faktory bHLH metabolismus MeSH
- transkripční faktory paired box genetika metabolismus MeSH
- tretinoin farmakologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
Anatomically widely different designs of animal eyes have long been thought to arise independently multiple times during evolution. This morphology-based view has been challenged by the identification of a highly conserved transcription factor Pax6 that plays a key role in eye development in both flies and mammals. The origin of Pax genes predates the origin of eyes and the nervous system since a PaxB-like gene, belonging to the Pax2/5/8 gene subfamily, was identified in sponge lacking nervous system. Structurally similar PaxB gene is implicated in visual system development in jellyfish, the most basal organism possessing complex eyes. The widespread use of Pax genes in the genetic program underlying eye formation throughout the animal kingdom raises a question why certain transcription factors have been frequently redeployed to build eyes. A model is proposed that provides a plausible explanation for the apparently ancient role of Pax genes in eye evolution.
