ABSTRACT
Using the rabbit corneal epithelial cell line RCE1(5T5) as a model, we analyzed three differentiation stages, distinguished on basis to the growth state of cultured cells and after studying the expression of transcription factors such as Oct4, Pax6 and ΔNp63α, selected differentiation markers, and signaling or epigenetic markers such as Notch receptors and Prdm3. Namely, proliferative non-differentiated cells, committed cells, and cells that constitute a stratified epithelium with a limbal epithelial-like structure. RNAseq based transcriptome analysis showed that 4891 genes were differentially expressed among these stages displaying distinctive gene signatures: proliferative cells had 1278 genes as gene signature, and seem to be early epithelial progenitors with an Oct4+, KLF4+, Myc+, ΔNp63α+, ABCG2+, Vimentin+, Zeb1+, VANGL1+, Krt3-, Krt12- phenotype. Committed cells had a gene signature with 417 genes and displayed markers indicative of the beginning of corneal differentiation, and genes characteristic of proliferative cells; we found the possible participation of Six3 and Six4 transcription factors along this stage. The third stage matches with a stratified corneal epithelium (gene signature comprising 979 genes) and is typified by an increase in the expression of WNT10A and NOTCH 2 and 3 signaling and Cux1 transcription factor, besides Pax6, KLF4 or Sox9. The differentiated cells express about 50% of the genes that belong to the Epidermal Differentiation Complex (EDC). Analysis of the differences between corneal epithelium and epidermis could be crucial to understand the regulatory mechanisms that lead to the expression of the differentiated phenotype.
Subject(s)
Cell Differentiation/physiology , Epithelium, Corneal/cytology , Transcriptome/physiology , Animals , Cells, Cultured , Epithelium, Corneal/metabolism , Fluorescent Antibody Technique, Indirect , Gene Expression Regulation/physiology , Octamer Transcription Factor-3/genetics , PAX6 Transcription Factor/genetics , Protein Interaction Maps , Rabbits , Real-Time Polymerase Chain Reaction , Transcription Factors/genetics , Vimentin/geneticsABSTRACT
BACKGROUND: Mutations in the BRCA1 and BRCA2 genes constitute a risk factor for breast cancer development. BRCA mutation research has been an active field since the discovery of the genes, and new mutations in both genes are constantly described and classified according to several systems. AIM: We intend to provide an overview of the current state of BRCA1 and BRCA2 mutation description and classification. We wanted to know whether there was a trend towards a more frequently described mutation type and what the proportion of pathogenic mutations was. RESULTS: We found that, although new mutations are described each year as reflected in current database records, very few of them are reported in papers. Classification systems are highly heterogeneous and a consensus among them is still under development. Regarding their function, a large number of mutations are yet to be analyzed, a very complex task, due to the great number of possible variations and their diverse effect in the BRCA gene functions. After individual analysis, many variants of unknown significance turn out to be pathogenic, and many can disrupt interactions with other proteins involved in mechanisms such as DNA damage repair pathways. Recent data suggest that looking for mutation patterns or combinations would shed a wider light on BRCA-derived cancer susceptibility in the upcoming years.
Subject(s)
BRCA1 Protein/genetics , BRCA2 Protein/genetics , Mutation , Breast Neoplasms/genetics , DNA Damage/genetics , DNA Repair/genetics , Female , Genetic Predisposition to Disease , HumansABSTRACT
ResumenLos especímenes silvestres de Vanilla planifolia G. Jack forman parte del acervo genético primario, los cuales solo se han reportado en Oaxaca, México. Por ello se evaluó la distribución de esta especie con el objetivo de ubicar y describir características ecológicas en zonas potenciales de distribución. La metodología empleada consistió de cuatro etapas: 1) Elaboración de una base de datos con registros de herbario;2) Construcción de la distribución potencial basado en los registros históricos de herbario para la especie, mediante el modelo de máxima entropía (Maxent), con el uso de 22 variables bioclimáticas como predictoras; 3) Realización de búsquedas sistemáticas de individuos in situ con base en los registros de herbario y las áreas de distribución potencial en 24 municipios, para conocer la situación y la distribución del hábitat actual, y 4) Descripción mediante factores ambientales de los nichos ecológicos potenciales generados por MaxEnt. La revisión de las colecciones de herbarios reportó un total de 18 registros de V. planifolia, comprendidos entre 1939 y 1998.La búsqueda sistemática de individuos en campo ubicó 28 plantas distribuidas en 12 sitios sobre 95 364 Km2. Las variables que contribuyeron con mayor valor porcentual para determinar la estimación del modelo de distribución potencial en vainilla son precipitación del periodo más lluvioso (61.9 %), régimen de humedad del suelo (23.4 %) y precipitación del cuatrimestre más lluvioso (8.1 %). El hábitat potencial de la especie se distribuyó en cuatro zonas; trópico húmedo del golfo de México, templado húmedo, trópico húmedo y templado húmedo del pacifico. La precipitación anual osciló de 2 500 a 4 000 mm, con lluvias en verano y porcentaje de precipitación invernal de 5 a 10 %. El régimen de humedad y clima predominantes fueron údico tipo I (330 a 365 días de humedad) y cálido húmedo (Am/A(C) m). Las plantas se ubicaron en altitudes de 200 a 1 190 msnm, en laderas accidentadas, que por lo general están al pie de sistemas montañosos de 1 300 a 2 500 metros de altitud. En condiciones naturales la distribución de la especie no se limita a selva alta perennifolia, dado que se ubicó en bosque mesófilo de montaña y bosque tropical perennifolio. La ubicación de nuevos especímenes de V. planifolia en condiciones silvestres reduce un 66 % del área potencial de distribución, y la fragmenta, al pasar de ser una zona continua a convertirse en tres zonas geográficamente separadas. La reducción del hábitat se debió a un aumento en el número de plantas ubicadas, lo que define las condiciones ambientales a un nivel más exacto. Por lo anterior, se pueden emprender o diseñar acciones de conservación enfocadas a áreas más específicas dentro del estado de Oaxaca, México.
AbstractWild specimens of Vanilla planifolia represent a vital part of this resource primary gene pool, and some plants have only been reported in Oaxaca, Mexico. For this reason, we studied its geographical distribution within the state, to locate and describe the ecological characteristics of the areas where they have been found, in order to identify potential areas of establishment. The method comprised four stages: 1) the creation of a database with herbarium records, 2) the construction of the potential distribution based on historical herbarium records for the species, using the model of maximum entropy (MaxEnt) and 22 bioclimatic variables as predictors; 3) an in situ systematic search of individuals, based on herbarium records and areas of potential distribution in 24 municipalities, to determine the habitat current situation and distribution; 4) the description of the environmental factors of potential ecological niches generated by MaxEnt. A review of herbarium collections revealed a total of 18 records of V. planifolia between 1939 and 1998. The systematic search located 28 plants distributed in 12 sites in 95 364 Km2. The most important variables that determined the model of vanilla potential distribution were: precipitation in the rainy season (61.9 %), soil moisture regime (23.4 %) and precipitation during the four months of highest rainfall (8.1 %). The species potential habitat was found to be distributed in four zones: wet tropics of the Gulf of Mexico, humid temperate, humid tropical, and humid temperate in the Pacific. Precipitation oscillated within the annual ranges of 2 500 to 4 000 mm, with summer rains, and winter precipitation as 5 to 10 % of the total. The moisture regime and predominating climate were udic type I (330 to 365 days of moisture) and hot humid (Am/A(C) m). The plants were located at altitudes of 200 to 1 190 masl, on rough hillsides that generally make up the foothills of mountain systems, with altitudes of 1 300 to 2 500 masl. In natural conditions, distribution of the species is not limited to high evergreen forests, since it was also found in mountain mesophyll and tropical evergreen forests. The location of new specimens of V. planifolia in its wild condition reduces the potential distribution area by 66 %. This area is fragmented into three geographically separated areas. Habitat reduction was due to the increased number of located plants that define the environmental conditions into a more accurate level. Conservation actions can thus be designed and implemented, focusing on more specific areas within the state of Oaxaca, Mexico.
Subject(s)
Vanilla/classification , Seasons , Biodiversity , Geography , MexicoABSTRACT
Wild specimens of Vanilla planifolia represent a vital part of this resource primary gene pool, and some plants have only been reported in Oaxaca, Mexico. For this reason, we studied its geographical distribution within the state, to locate and describe the ecological characteristics of the areas where they have been found, in order to identify potential areas of establishment. The method comprised four stages: 1) the creation of a database with herbarium records, 2) the construction of the potential distribution based on historical herbarium records for the species, using the model of maximum entropy (MaxEnt) and 22 bioclimatic variables as predictors; 3) an in situ systematic search of individuals, based on herbarium records and areas of potential distribution in 24 municipalities, to determine the habitat current situation and distribution; 4) the description of the environmental factors of potential ecological niches generated by MaxEnt. A review of herbarium collections revealed a total of 18 records of V. planifolia between 1939 and 1998. The systematic search located 28 plants distributed in 12 sites in 95 364 Km(2). The most important variables that determined the model of vanilla potential distribution were: precipitation in the rainy season (61.9 %), soil moisture regime (23.4 %) and precipitation during the four months of highest rainfall (8.1 %). The species potential habitat was found to be distributed in four zones: wet tropics of the Gulf of Mexico, humid temperate, humid tropical, and humid temperate in the Pacific. Precipitation oscillated within the annual ranges of 2 500 to 4 000 mm, with summer rains, and winter precipitation as 5 to 10 % of the total. The moisture regime and predominating climate were udic type I (330 to 365 days of moisture) and hot humid (Am/A(C) m). The plants were located at altitudes of 200 to 1 190 masl, on rough hillsides that generally make up the foothills of mountain systems, with altitudes of 1 300 to 2 500 masl. In natural conditions, distribution of the species is not limited to high evergreen forests, since it was also found in mountain mesophyll and tropical evergreen forests. The location of new specimens of V. planifolia in its wild condition reduces the potential distribution area by 66 %. This area is fragmented into three geographically separated areas. Habitat reduction was due to the increased number of located plants that define the environmental conditions into a more accurate level. Conservation actions can thus be designed and implemented, focusing on more specific areas within the state of Oaxaca, Mexico.
