Abstract
DAPI staining of the metaphase chromosomes pretreated with barium hydroxide generates a C-like banding pattern. In this work a protocol for visualizing similar pattern at the synaptonemal complex (SC) spreads after immunostaining is suggested. This method was used to visualize centromeric and sex heterochromatin at the SC spreads of guppy fish (Poecilia reticulata Peters, 1859). The efficiency of this method was further confirmed at SC spreads of the northern red-backed vole (Myodes rutilus (Pallas, 1779)), the guinea pig (Cavia porcellus (Linnaeus, 1758)), and the pigmy shrew (Sorex minutus Linnaeus, 1766).
Highlights
Immunofluorescent analysis of synaptonemal complexes (SC) is widely performed in medical and comparative cytogenetics to study chromosome synapsis and recombination in humans with chromosomal abnormalities (Solari 1999) as well as in various plant and animal species (Anderson et al 1999; Basheva et al 2010; Borodin et al 2008)
Guinea pig and pigmy shrew, the centromeres were marked by antibodies and DAPI signals
DAPI-stained pachytene cells untreated with barium hydroxide produced a unique banding pattern for each chromosome. This banding pattern is visible even without image contrast adjustment, and bright bands correspond to centromere antibody signals (Fig. 1a)
Summary
Immunofluorescent analysis of synaptonemal complexes (SC) is widely performed in medical and comparative cytogenetics to study chromosome synapsis and recombination in humans with chromosomal abnormalities (Solari 1999) as well as in various plant and animal species (Anderson et al 1999; Basheva et al 2010; Borodin et al 2008). In this study a simple protocol was designed to obtain C-like banding on SC spreads and visualize centromeres without the use of antibodies. The DAPI staining after barium hydroxide treatment was shown to visualize C-like banding on the metaphase chromosomes of fish (Russo, Rocco et al 1999), plants (Barros e Silva and Guerra 2010), insects (Bella and Gosálvez 1991) and human (Heng and Tsui 1993).
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