Evolutionary dynamics of repetitive DNA in semaprochilodus (characiformes, prochilodontidae): a fish model for sex chromosome differentiation.

Distribution of 6 microsatellites and 5 transposable elements on the chromosomes of Semaprochilodus taeniurus and S. insignis, commonly referred to as Jaraqui, was performed using their physical mapping with fluorescence in situ hybridization. In this study, we aim to understand the evolutionary dynamics in genomes of S. taeniurus and S. insignis by comparing the position, abundance and contribution of the repetitive sequences in the origins and differentiation of a ZZ/ZW sex chromosome system in S. taeniurus. Results revealed that distribution patterns of repetitive DNAs along the chromosomes varied considerably. Hybridization signals were observed on several autosomes in both species; however, in S. taeniurus genome, the repetitive sequences were more abundant. In addition, large clusters of known repetitive sequences were detected in sex chromosomes of S. taeniurus. This observation is notable because the accumulation of repetitive DNAs could reflect the degradation of genetic activities and the differentiation of protosex chromosomes, evolving into the heteromorphic ZW pair observed in S. taeniurus.

Cryptic diversity in the mtDNA of the ornamental fish Carnegiella strigata.

Mitochondrial DNA ( mtDNA) sequences of the marbled hatchetfish Carnegiella strigata, an ornamental fish exported from the Negro River, was examined to determine its genetic diversity and population structure in blackwater rivers (Negro and Uatumã Rivers) in the central Amazon. Analyses of a 646 bp fragment of the ATPase 6/8 mtDNA gene revealed two monophyletic lineages of C. strigata with considerable genetic distance between them (10-12%), suggesting that these lineages should not be considered a single stock. Furthermore, there were strong differences in the geographical distribution of the lineages. These results indicate a past association between drainages of the Negro and Uatumã Rivers. They also suggest that, in the Negro River, its main tributary, the Branco River, may act as a geographical barrier and potentially an ecological barrier between populations of the middle and lower portions of the river.

Cytogenetics of Synbranchiformes: a comparative analysis of two Synbranchus Bloch, 1795 species from the Amazon.

Synbranchidae belongs to the Synbranchiformes and occurs in Africa, Asia, Australia, Mexico, and Central and South America. This family comprises four genera: Synbranchus, Ophisternon, Monopterus, and Macrotrema. Only two are known from the neotropical region, Ophisternon and Synbranchus. According to current classification, Synbranchus has three valid species: S. marmoratus (Bloch 1795), S. madeirae (Rosen and Rumney 1972), and S. lampreia (Favorito, Zanata and Assumpção 2005). Thus the present research is aimed to cytogenetically characterize (by classical and molecular methods) two syntopic species-S. aff. lampreia and S. madeirae-from the central Amazon basin to validate the taxonomy of both species and provide a revisionary discussion on the cytogenetics of Synbranchiformes. Synbranchus aff. lampreia was found to possess 2n = 44 chromosomes (6 m + 2st + 36a, NF = 50), while S. madeirae had 2n = 46 chromosomes (6 m + 2st + 38a, NF = 52). Constitutive heterochromatin was dominant in the centromeric and terminal regions of most of the chromosomes in both species, although the precise distribution patterns were species-specific. The nucleolar organizing region was single in S. aff. lampreia and multiple in S. madeirae, as indicated by both AgNO(3) and hybridization using 18S rDNA probes. The 5S rDNA sites were located interstitially on the long arms of an acrocentric pair in both species, and the telomeric probe did not show any interstitial sites in either species. These data indicate the occurrence of interspecific karyotypic variability in Synbranchus and suggest that taxonomic review for this genus is necessary.

Comparative analysis of skin surface lipids of the labia majora, inner thigh, and forearm.

The purpose of this study was to develop a procedure for the collection of skin surface corneocyte lipids from the semioccluded and intimate regions of the labia majora and inner thigh of women, to evaluate the polar and nonpolar composition, and to compare the distribution of the lipid classes relative to a collection of lipids from the forearm. The solvent system of ethanol-cyclohexane was well tolerated across all sites. While the yield of polar lipids was similar across all 3 sites, there were only marginal differences in the relative abundance of ceramides, a class of lipids closely associated with skin barrier activity. The yield of neutral lipids was significantly less for the labia majora and was associated with a reduced yield of wax esters, triglycerides and free fatty acids, likely associated with reduced sebaceous gland activity. Factors that may contribute to an inferior skin barrier activity for the labia majora are discussed and suggest a possible deficiency of ω-6 fatty acid linked to the sphingosine base of ceramide EOS.

CompCytogen of two of the smallest Amazonian fishes: Fluviphylax simplex Costa, 1996 and Fluviphylax zonatus Costa, 1996 (Cyprinodontiformes, Poeciliidae).

The genus Fluviphylax Whitley, 1965is comprized of five valid species (Fluviphylax pygmaeus Myers et Carvalho, 1955, Fluviphylax zonatus, Fluviphylax simplex, Fluviphylax obscurus Costa, 1996,and Fluviphylax palikur Costa et Le Bail, 1999), which are endemic to the Amazon region. These fishes are the smallest known South American vertebrates and among the smallest know vertebrates on Earth. All species but the type Fluviphylax pygmaeus have been described in late 1990's, and much remains unknown about the biology, taxonomy and systematics of this group of fishes. The aims of the present study were to establish the diploid and haploid number of Fluviphylax zonatus and Fluviphylax simplex, and to find species-specific markers for the discrimination of taxa. The diploid number for both species was 48 chromosomes, with no sex chromosome heteromorphism. Fluviphylax zonatus exhibited the karyotypic formula 4m+8sm+22st+14a and FN=82, and Fluviphylax simplex exhibited 4m+16sm+18st+10a and FN=86. The determination of the total mean length of the chromosomes and their grouping into five size classes demonstrated different chromosome composition of the two species. This difference was further supported by the distribution of constitutive heterochromatin. The meiotic analysis revealed 24 bivalents in both species, but Fluviphylax zonatus exhibited chromosomes with late pairing of the telomeric portions in the pachytene. These data reveal that cytogenetic characterization is useful and important for the discrimination of these species. Our study further indicates that this method could be employed in the analysis of other species of small fishes that are difficult to distinguish using traditional morphological traits or are morphologically cryptic.

Variability of 18S rDNA locus among Symphysodon fishes: chromosomal rearrangements.

Three species of cichlids belonging to the genus Symphysodon have demonstrated interspecific and intraspecific variation in nucleolus organizer regions (NOR) detected with silver nitrate. In order to understand the evolution of this marker in the genus, the structural variability of these sequences in mitotic chromosomes from Symphysodon aequifasciatus, Symphysodon discus and Symphysodon haraldi was investigated using both silver nitrate impregnation and hybridization of the 18S rRNA gene probe. For the three species, the two markers were intraspecifically and interspecifically variable both in the number and in the size of the sites. This polymorphism may stem from duplications and translocations, which suggests that structural chromosome rearrangements effectively act in the karyoevolution of wild Symphysodon species and may have favoured the adaptability of these fishes to diverse aquatic environments in the Amazon.

Intriguing evidence of translocations in Discus fish (Symphysodon, Cichlidae) and a report of the largest meiotic chromosomal chain observed in vertebrates.

As part of a program to understand the genetics of Amazonian ornamental fish, classical cytogenetics was used to analyze Symphysodon aequifasciatus, S. discus and S. haraldi, popular and expensive aquarium fishes that are endemic to the Amazon basin. Mitotic analyses in Symphysodon have shown some odd patterns compared with other Neotropical cichlids. We have confirmed that Symphysodon species are characterized by chromosomal diversity and meiotic complexity despite the fact that species share the same diploid number 2n=60. An intriguing meiotic chromosomal chain, with up to 20 elements during diplotene/diakinesis, was observed in S. aequifasciatus and S. haraldi, whereas S. discus only contains typical bivalent chromosomes. Such chromosomal chains with a high number of elements have not been observed in any other vertebrates. We showed that the meiotic chromosomal chain was not sex related. This observation is unusual and we propose that the origin of meiotic multiples in males and females is based on a series of translocations that involved heterochromatic regions after hybridization of ancestor wild Discus species.

Comparative cytogenetic analysis of the genus symphysodon (discus fishes, cichlidae): chromosomal characteristics of retrotransposons and minor ribosomal DNA.

As part of a genetic screening program for wild Discus fishes, we analyzed karyotypes and cytogenetic characteristics of Symphysodon aequifasciatus, S. discus and S. haraldi using C-banding and fluorescent in situ hybridization (FISH) with the Rex3 retrotransposon and 5S rDNA probes in mitotic and meiotic chromosomes. In the 3 species, diploid chromosome number was 2n = 60 and karyotypes contained predominantly meta-submetacentric chromosomes. C-banding showed blocks of constitutive heterochromatin mainly in the pericentromeric region. Physical mapping of repetitive 5S rDNA sequences and Rex3 retrotransposons in mitotic and meiotic chromosomes showed partial colocalization of constitutive heterochromatin and repetitive elements. Correlations among the accumulation of repetitive elements, heterochromatinization and chromosome rearrangements have been hypothesized to explain the karyotype differentiation in the Symphysodon genus. The role of repetitive elements in adaptation to highly diverse habitats, as well as in the generation of the phenotypic and genetic variability found in wild Discus populations, needs to be further investigated.

Cyclic AMP response to recombinant human relaxin by cultured human endometrial cells--a specific and high throughput in vitro bioassay.

A specific and high throughput 96-well format bioassay for recombinant human relaxin (rhRLX) has been developed using human endometrial cells (NHE cells). rhRLX caused a time- and dose-dependent stimulation of cyclic AMP (cAMP) with 1/2 maximal activity of 3.56 +/- 0.65 ng/ml (n = 30). The range of the standard curve was 0.39 to 25 ng/ml with interplate precision of 17 and 22% CV for high and low controls respectively. The cAMP response requires forskolin and 3-isobutyl-1-methylxanthine, and is enhanced by prostaglandin E2 and F2 alpha. The NHE cells do not respond to A or B chains of rhRLX, or a whole array of hormones. Preincubation of rhRLX with specific monoclonal antibody completely abolished the cAMP response. This bioassay has been used to determine the biological activity of several manufactured lots of recombinant human relaxin.