Abstract
BackgroundDifferent 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.ResultsThe 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.ConclusionHere, 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.
Highlights
Different moderrn methodologies are presently available to analyze meiotic chromosomes
We demonstrate that the new methodproposed here is effective for species of several groups and even for use in combination with other methods, such as immunodetection of the SYCP3 protein with a specific antibody and co-detection of DNA sequences by fluorescence in situ hybridization (FISH)
Meiotic preparations from different species of fishes such as Oreochromis niloticus, Eigenmannia sp2, Astyanax paranae, and Characidium gomesi were obtained with application of the surface-spreading protocol proposed in this study (Figure 1), highlighting the replicability of the method
Summary
Different moderrn methodologies are presently available to analyze meiotic chromosomes. Specific events are required to promote genetic diversity and ensure the segregation of homologous chromosomes. These events, including synapsis and meiotic recombination between homologous chromosomes, cause sequential structural changes in chromosomes [1]. Studies on these structural elements can provide important information about pairing and synapsis processes involving homologous chromosomes [2], as well as about the evolution of protein components of the synaptonemal complex (SC) among vertebrates [3]. Seven proteins are described as part of the SC: SYCP2 and SYCP3, forming the lateral elements [7,8]; SYCP1, which is the unique protein that joins the transversal elements [9] and four small proteins, SYCE1, SYCE2, SYCE3 and Tex, which are specific to the central element [10,11,12]
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