title: Surface-spreading technique of meiotic cells and immunodetection of synaptonemal complex proteins in teleostean fishes.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors: Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, Distrito de Rubião Junior, s/n, 18618-970 Botucatu, SP Brazil.
authors: Cristian
authors: C
authors: Araya-Jaime
authors: Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, Distrito de Rubião Junior, s/n, 18618-970 Botucatu, SP Brazil.
authors: Érica Alves
authors: ÉA
authors: Serrano
authors: Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, Distrito de Rubião Junior, s/n, 18618-970 Botucatu, SP Brazil.
authors: Duílio Mazzoni Zerbinato
authors: DM
authors: de Andrade Silva
authors: Department of Biological Sciences, Faculty of Science Hokkaido University, Sapporo, 060-0810 Japan.
authors: Masakane
authors: M
authors: Yamashita
authors: South Ehime Fisheries Research Center, Ehime University, Matsuyama, Ehime 798-4292 Japan.
authors: Toshiharu
authors: T
authors: Iwai
authors: Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, Distrito de Rubião Junior, s/n, 18618-970 Botucatu, SP Brazil.
authors: Cláudio
authors: C
authors: Oliveira
authors: Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista, Distrito de Rubião Junior, s/n, 18618-970 Botucatu, SP Brazil.
authors: Fausto
authors: F
authors: Foresti
keywords: Co-detection; FISH; Meiosis; Method; SYCP3
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