Cargando…

Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes

BACKGROUND: Different moderrn methodologies are presently available to analyze meiotic chromosomes. These methods permit investigation of the behavior of chromosomes in the normal complement and of sex and B chromosomes, two special types of chromosomes that are associated with the A complement and...

Descripción completa

Detalles Bibliográficos
Autores principales: Araya-Jaime, Cristian, Serrano, Érica Alves, de Andrade Silva, Duílio Mazzoni Zerbinato, Yamashita, Masakane, Iwai, Toshiharu, Oliveira, Cláudio, Foresti, Fausto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4312464/
https://www.ncbi.nlm.nih.gov/pubmed/25642289
http://dx.doi.org/10.1186/s13039-015-0108-9
_version_ 1782355125348597760
author Araya-Jaime, Cristian
Serrano, Érica Alves
de Andrade Silva, Duílio Mazzoni Zerbinato
Yamashita, Masakane
Iwai, Toshiharu
Oliveira, Cláudio
Foresti, Fausto
author_facet Araya-Jaime, Cristian
Serrano, Érica Alves
de Andrade Silva, Duílio Mazzoni Zerbinato
Yamashita, Masakane
Iwai, Toshiharu
Oliveira, Cláudio
Foresti, Fausto
author_sort Araya-Jaime, Cristian
collection PubMed
description BACKGROUND: Different moderrn methodologies are presently available to analyze meiotic chromosomes. These methods permit investigation of the behavior of chromosomes in the normal complement and of sex and B chromosomes, two special types of chromosomes that are associated with the A complement and are present in many organisms, including fishes. However, meiotic studies are still scarce in fishes, considering the wide number of species in this group.. Here, we describe a new protocol for the visualization of the synaptonemal complex in spermatocytes and oocytes of fishes and to the sequential use of the technique with other procedures and techniques such as immunodetection of the synaptonemal complex protein with a specific antibody and co-detection of DNA sequences by FISH. RESULTS: The meiotic surface-spreading protocol used in the present proposal worked well in representative species of four fish orders and was useful in obtaining good results even in small specimens. Fish-specific antibodies and commercial products worked similarly well to detect synaptonemal complex (SC) proteins. The sequential application of fluorescence in situ hybridization using specific probes showed clear signals associated with the SC structures identified by immunostaining. CONCLUSION: Here, we provide a useful and applicable immunofluorescent protocol for the visualization of synaptonemal complex proteins in the meiotic cells of fishes in surface-spreading preparations. Furthermore, this technique allows for the sequential application of other cytogenetic procedures.
format Online
Article
Text
id pubmed-4312464
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-43124642015-02-01 Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes Araya-Jaime, Cristian Serrano, Érica Alves de Andrade Silva, Duílio Mazzoni Zerbinato Yamashita, Masakane Iwai, Toshiharu Oliveira, Cláudio Foresti, Fausto Mol Cytogenet Methodology BACKGROUND: Different moderrn methodologies are presently available to analyze meiotic chromosomes. These methods permit investigation of the behavior of chromosomes in the normal complement and of sex and B chromosomes, two special types of chromosomes that are associated with the A complement and are present in many organisms, including fishes. However, meiotic studies are still scarce in fishes, considering the wide number of species in this group.. Here, we describe a new protocol for the visualization of the synaptonemal complex in spermatocytes and oocytes of fishes and to the sequential use of the technique with other procedures and techniques such as immunodetection of the synaptonemal complex protein with a specific antibody and co-detection of DNA sequences by FISH. RESULTS: The meiotic surface-spreading protocol used in the present proposal worked well in representative species of four fish orders and was useful in obtaining good results even in small specimens. Fish-specific antibodies and commercial products worked similarly well to detect synaptonemal complex (SC) proteins. The sequential application of fluorescence in situ hybridization using specific probes showed clear signals associated with the SC structures identified by immunostaining. CONCLUSION: Here, we provide a useful and applicable immunofluorescent protocol for the visualization of synaptonemal complex proteins in the meiotic cells of fishes in surface-spreading preparations. Furthermore, this technique allows for the sequential application of other cytogenetic procedures. BioMed Central 2015-01-27 /pmc/articles/PMC4312464/ /pubmed/25642289 http://dx.doi.org/10.1186/s13039-015-0108-9 Text en © Araya-Jaime et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology
Araya-Jaime, Cristian
Serrano, Érica Alves
de Andrade Silva, Duílio Mazzoni Zerbinato
Yamashita, Masakane
Iwai, Toshiharu
Oliveira, Cláudio
Foresti, Fausto
Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title_full Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title_fullStr Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title_full_unstemmed Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title_short Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
title_sort surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4312464/
https://www.ncbi.nlm.nih.gov/pubmed/25642289
http://dx.doi.org/10.1186/s13039-015-0108-9
work_keys_str_mv AT arayajaimecristian surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT serranoericaalves surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT deandradesilvaduiliomazzonizerbinato surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT yamashitamasakane surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT iwaitoshiharu surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT oliveiraclaudio surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes
AT forestifausto surfacespreadingtechniqueofmeioticcellsandimmunodetectionofsynaptonemalcomplexproteinsinteleosteanfishes