Use of chromosome microdissection in fish molecular cytogenetics

Henning, Frederico; Трифонов, Владимир; Almeida-Toledo, Lurdes Foresti de

doi:10.1590/s1415-47572008000200022

Cited by 16 publications

(17 citation statements)

References 22 publications

(19 reference statements)

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“…from the region of Bahia and the eastern Amazon suggest the occurrence of the XX/ XY 1 Y 2 and ZZ/ZW systems, respectively Milhomem et al, 2008]. This indicates a likely independent origin, as occurs in species of Eigenmannia (Gymnotiformes) distributed in 3 different hydrographic basins in South America [Henning et al, 2008]. The meiotic analysis should be considered fundamental for a precise characterization of sex chromosomes, their behavior during cell division and formation of gametes.…”

Section: Discussionmentioning

confidence: 96%

Repetitive DNA and Meiotic Behavior of Sex Chromosomes in Gymnotus pantanal (Gymnotiformes, Gymnotidae)

Silva

Matoso²,

Vicari³

et al. 2011

Cytogenet Genome Res

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Neotropical fishes have a low rate of chromosome differentiation between sexes. The present study characterizes the first meiotic analysis of sex chromosomes in the order Gymnotiformes. Gymnotus pantanal – females had 40 chromosomes (14m/sm, 26st/a) and males had 39 chromosomes (15m/sm, 24st/a), with a fundamental number of 54 – showed a multiple sexual determination chromosome system of the type X₁X₁X₂X₂/X₁X₂Y. The heterochromatin is restricted to centromeres of all chromosomes of the karyotype. The meiotic behavior of sex chromosomes involved in this system in males is from a trivalent totally pared in the pachytene stage, with a high degree of similarity. The cells of metaphase II exhibit 19 and 20 chromosomes, normal disjunction of sex chromosomes and the formation of balanced gametes with 18 + Y and 18 + X₁X₂ chromosomes, respectively. The small amount of heterochromatin and repetitive DNA involved in this system and the high degree of chromosome similarity indicated a recent origin of the X₁X₁X₂X₂/X₁X₂Y system in G. pantanal and suggests the existence of a simple ancestral system with morphologically undifferentiated chromosomes.

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Section: Discussionmentioning

confidence: 96%

Repetitive DNA and Meiotic Behavior of Sex Chromosomes in Gymnotus pantanal (Gymnotiformes, Gymnotidae)

Silva

Matoso²,

Vicari³

et al. 2011

Cytogenet Genome Res

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“…However, little is known about the molecular composition of the DNA specifically present in fish B chromosomes [Jesus and Moreira-Filho, 2003]. Indeed, probes derived from chromosome microdissection have been successfully used in studies of fish chromosomes, as in the analysis of homologous chromosomes in Eigenmannia [Henning et al, 2008] and Cyprinid fish [Ráb et al, 2008], of sex chromosomes in Triportheus [Diniz et al, 2008] and Characidium [Machado et al, 2011;Pazian et al, 2013], and supernumerary chromosomes in Prochilodus lineatus [Voltolin et al, 2010]. However, still little is known about the molecular composition of the B chromosomes among fishes as a whole.…”

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confidence: 99%

Common Descent of B Chromosomes in Two Species of the Fish Genus Prochilodus (Characiformes, Prochilodontidae)

Voltolin

Alves²,

Senhorini

et al. 2013

Cytogenet Genome Res

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To ascertain the origin of B chromosomes in 2 fish species of the genus Prochilodus, i.e. P. lineatus and P. nigricans, we microdissected them and generated B-specific DNA probes. These probes were used to perform chromosome painting in both species and in 3 further ones belonging to the same genus (P. argenteus, P. brevis and P. costatus). Both probes hybridized with the B chromosomes in P. lineatus and P. nigricans, but with none of the chromosomes in the 5 species. This indicates that the B chromosomes have low similarity with DNAs located in the A chromosomes and suggests the possibility that the B chromosomes in the 2 species have a common origin. The most parsimonious explanation would imply intergeneric hybridization in an ancestor of P. lineatus and P. nigricans yielding the B chromosome as a byproduct, which remained in these 2 species after their phylogenetic origin, but was perhaps lost in other Prochilodus species. This hypothesis predicts that B chromosomes are old genomic elements in this genus, and this could be tested once a species from a relative genus would be found showing homology of its A chromosomes with the B-probes employed here, through a comparison of B chromosome DNA sequences with those in the A chromosomes of this other species.

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“…Studies have reported the need for large numbers of micromanipulated chromosomes to construct a probe (approximately 20). However, some authors have pointed out that the construction of probes with a lower number of chromosomes is possible (Guan et al 1994, Engelen et al 1998, Henning et al 2008. Christian et al (1999) described a protocol to generate probes from a single micromanipulated chromosome.…”

Section: Discussionmentioning

confidence: 99%

“…In this sense, it is necessary to improve the time and temperature of annealing and extension for the enzymes. As the latter are highly processive at high temperatures, it is recommended to use the ramp rate between the annealing and extension temperatures (Henning et al 2008). Another relevant factor in the amplification step is the primer concentration, as limiting primer levels may decrease the amplification rate in the last cycles of the PCR procedure (Czerny 1996).…”

Section: Discussionmentioning

confidence: 99%

Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques

Passamani

Carvalho

Soares

2017

An. Acad. Bras. Ciênc.

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Chromosome-specific probes have been widely used in molecular cytogenetics, being obtained with different methods. In this study, a reproducible protocol for construction of chromosome-specific probes is proposed which associates in situ amplification (PRINS), micromanipulation and degenerate oligonucleotide-primed PCR (DOP-PCR). Human lymphocyte cultures were used to obtain metaphases from male and female individuals. The chromosomes were amplified via PRINS, and subcentromeric fragments of the X chromosome were microdissected using microneedles coupled to a phase contrast microscope. The fragments were amplified by DOP-PCR and labeled with tetramethyl-rhodamine-5-dUTP. The probes were used in fluorescent in situ hybridization (FISH) procedure to highlight these specific regions in the metaphases. The results show one fluorescent red spot in male and two in female X chromosomes and interphase nuclei.

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Use of chromosome microdissection in fish molecular cytogenetics

Cited by 16 publications

References 22 publications

Repetitive DNA and Meiotic Behavior of Sex Chromosomes in <i>Gymnotus pantanal</i> (Gymnotiformes, Gymnotidae)

Repetitive DNA and Meiotic Behavior of Sex Chromosomes in <i>Gymnotus pantanal</i> (Gymnotiformes, Gymnotidae)

Common Descent of B Chromosomes in Two Species of the Fish Genus <b><i>Prochilodus</i></b> (Characiformes, Prochilodontidae)

Protocol for chromosome-specific probe construction using PRINS, micromanipulation and DOP-PCR techniques

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