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Author | Mireia Sole; Joan Blanco; Debora Gil; G. Fonseka; Richard Frodsham; Francesca Vidal; Zaida Sarrate | ||||
Title | Noves perspectives en l estudi de la territorialitat cromosomica de cel·lules germinals masculines: estudis tridimensionals | Type | Journal | ||
Year | 2017 | Publication | Biologia de la Reproduccio | Abbreviated Journal | JBR |
Volume | 15 | Issue | Pages | 73-78 | |
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Abstract | In somatic cells, chromosomes occupy specific nuclear regions called chromosome territories which are involved in the
maintenance and regulation of the genome. Preliminary data in male germ cells also suggest the importance of chromosome territoriality in cell functionality. Nevertheless, the specific characteristics of testicular tissue (presence of different cell types with different morphological characteristics, in different stages of development and with different ploidy) makes difficult to achieve conclusive results. In this study we have developed a methodology to approach the threedimensional study of all chromosome territories in male germ cells from C57BL/6J mice (Mus musculus). The method includes the following steps: i) Optimized cell fixation to obtain an optimal preservation of the three-dimensionality cell morphology, ii) Chromosome identification by FISH (Chromoprobe Multiprobe® OctoChrome™ Murine System; Cytocell) and confocal microscopy (TCS-SP5, Leica Microsystems), iii) Cell type identification by immunofluorescence iv) Image analysis using Matlab scripts, v) Numerical data extraction related to chromosome features, chromosome radial position and chromosome relative position. This methodology allows the unequivocally identification and the analysis of the chromosome territories of all spermatogenic stages. Results will provide information about the features that determine chromosomal position, preferred associations between chromosomes, and the relationship between chromosome positioning and genome regulation. |
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ISSN | ISBN | 978-84-697-3767-5 | Medium | ||
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Notes | IAM; 600.096; 600.145 | Approved | no | ||
Call Number | Admin @ si @ SBG2017c | Serial | 2961 | ||
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