The X1X2Y sex chromosome system in the fish Hoplias malabaricus. I. G-, C- and chromosome replication banding

Hoplias malabaricus, a widely distributed neotropical fish (Central America to Argentina), may represent a group of distinct species showing diversified cytotypes with respect to chromosome number, morphology and sex systems. One of these karyotypic forms is characterized by an X₁X₁X₂X₂/X₁X₂Y sex chromosome system, with 2n= 40 and 39 chromosomes in females and males respectively. Analyses with G-, C- and chromosome replication banding permitted a better characterization of the sex chromosomes in this cytotype. The Y chromosome, unique in males, resulted from a translocation event between two biarmed chromosomes: one similar to chromosome 6 (X₁) and the other one similar to chromosome 20 (X₂), the latter corresponding to a probable identification. On the basis of the observed banding patterns, the Y chromosome may represent a stable dicentric, with an inactive centromere interstitially located on its long arm. The results are also related to a specific satellite DNA subfamily, previously characterized in Hoplias malabaricus, which appears to be associated with the X₁ chromosome.This revised version was published online in November 2005 with corrections to the Cover Date.     

A biodiversity approach in the neotropical Erythrinidae fish, Hoplias malabaricus. Karyotypic survey, geographic distribution of cytotypes and cytotaxonomic considerations

Hoplias malabaricus, a widely distributed neotropical freshwater fish, shows a conspicuous karyotypic diversification. An overview of this diversity is presented here comprising several Brazilian populations, and some others from Argentina, Uruguay and Surinam. Seven general cytotypes are clearly identified on the basis of their diploid number (2n=39 to 2n=42), chromosomal morphology and sex chromosome systems, which can be clustered into two major karyotypic groups. This clustering suggests that karyotype structure would be more informative than the diploid number regarding cytotype relationships in this fish group. While some cytotypes show a wide geographical distribution, some others appear to be endemic to specific hydrographic basins. Sympatric cytotypes can occur without detection of hybrid forms; this situation points to a lack of gene flow, a fact that is also reinforced by studies with genomic markers. The karyotypic data support the view that the nominal taxon H. malabaricus corresponds to a species complex comprising distinct evolutionary units, each with well-established chromosomal differences.     

An XX/XY sex microchromosome system in a freshwater turtle, Chelodina longicollis (Testudines: Chelidae) with genetic sex determination

Heteromorphic sex chromosomes are rare in turtles, having been described in only four species. Like many turtle species, the Australian freshwater turtle Chelodina longicollis has genetic sex determination, but no distinguishable (heteromorphic) sex chromosomes were identified in a previous karyotyping study. We used comparative genomic hybridization (CGH) to show that C. longicollis has an XX/XY system of chromosomal sex determination, involving a pair of microchromosomes. C-banding and reverse fluorescent staining also distinguished microchromosomes with different banding patterns in males and females in ∼70% cells examined. GTG-banding did not reveal any heteromorphic chromosomes, and no replication asynchrony on the X or Y microchromosomes was observed using replication banding. We conclude that there is a very small sequence difference between X and Y chromosomes in this species, a difference that is consistently detectable only by high-resolution molecular cytogenetic techniques, such as CGH. This is the first time a pair of microchromosomes has been identified as the sex chromosomes in a turtle species.     

Sex chromosome evolution in fish. II. Second occurrence of an X1X2Y sex chromosome system in Gymnotiformes

A multiple sex chromosome system of the X₁X₁X₂X₂:X₁X₂Y type is reported to occur in the fish species Brachyhypopomus pinnicaudatus (Gymnotiformes, Hypopomidae), being the second occurrence of this sex chromosome system in Gymnotiformes and the fifth among Neotropical freshwater fish. The possible origin of this system was hypothesized to be a centric fusion, which occurred in an ancestral form, of two medium-sized acrocentrics, giving origin to the metacentric neo-Y. Heterochromatic DAPI-positive regions were visualized in the pericentromeric region of all the chromosomes, including the Y-chromosome. In-situ hybridization with (TTAGGG) _n (all-human-telomeres probe) did not detect any telomeric interstitial regions (ITS), indicating a possible loss of terminal segments of the chromosomes involved in the neo-Y formation. This revised version was published online in July 2006 with corrections to the Cover Date.     

An XX/XY heteromorphic sex chromosome system in the Australian chelid turtle <Emphasis Type="Italic">Emydura macquarii</Emphasis>: A new piece in the puzzle of sex chromosome evolution in turtles

Chromosomal sex determination is the prevalent system found in animals but is rare among turtles. In fact, heteromorphic sex chromosomes are known in only seven of the turtles possessing genotypic sex determination (GSD), two of which correspond to cryptic sex microchromosomes detectable only with high-resolution cytogenetic techniques. Sex chromosomes were undetected in previous studies of Emydura macquarii, a GSD side-necked turtle. Using comparative genomic hybridization (CGH) and GTG-banding, a heteromorphic XX/XY sex chromosome system was detected in E. macquarii. The Y chromosome appears submetacentric and somewhat larger than the metacentric X, the first such report for turtles. CGH revealed a male-specific chromosomal region, which appeared heteromorphic using GTG-banding, and was restricted to the telomeric region of the p arm. Based on our observations and the current phylogeny of chelid turtles, we hypothesize that the sex chromosomes of E. macquarii might be the result of a translocation of an ancestral Y microchromosome as found in a turtle belonging to a sister clade, Chelodina longicollis, onto the tip of an autosome. However, in the absence of data from an outgroup, the opposite (fission of a large XY into an autosome and a micro-XY) is theoretically equally likely. Alternatively, the sex chromosome systems of E. macquarii and C. longicollis may have evolved independently. We discuss the potential causes and consequences of such putative chromosome rearrangements in the evolution of sex chromosomes and sex-determining systems of turtles in general.     

Ac-like transposons in populations of wild diploid Triticeae species: comparative analysis of chromosomal distribution

Data are presented on the intra- and interspecific differences/similarities in chromosomal patterns of Ac-like elements (hAT family) in ecologically contrasted populations of three Triticeae species - Aegilops speltoides, Triticum urartu, and Hordeum spontaneum. Application of original computer software made it possible to precisely map transposon clusters and to link them to known chromosomal markers (rDNA sites, centromeres, and heterochromatin regions). From our data we can specify the most visible features of Ac-like elements chromosomal distribution: preferential concentration in chromosomal proximal regions; high percentage of clusters on the border between euchromatin and heterochromatin; complementary chromosomal arrangement towards En/Spm transposons (CACTA); population-specific insertions into centromeres; more differences in total cluster numbers between populations of self-pollinated species than between populations of cross-pollinated species. The application of statistical simulation (Resampling) method to analysis of data indicates that ecology may play a certain role in dynamics of Ac-like elements. Comparison of real Ayala distances, as well as real chromosomal distribution of Ac-like elements in populations of two species with different mating systems with the same but randomly simulated parameters, revealed that non-random population structure in the Mediterranean floral zone suffers and becomes chaotic in the Irano-Turanian zone.     

Additional locus of rDNA sequence specific to the X chromosome of the liverwort, Marchantia polymorpha

The molecular cytogenetic organization of 17S ribosomal RNA genes (17S rDNA), a part of the 45S rDNA repeat, was investigated on the chromosomes of the liverwort Marchantia polymorpha using fluorescence in-situ hybridization (FISH). The numbers of 17S rDNA loci visualized in female and male chromosomes were ten and nine, respectively. This heterogeneous localization was due to the presence of an additional 17S rDNA locus on the X chromosome and its absence on the Y chromosome. The signal on the X chromosome covered almost the entire region of its long arm. The other nine signals were observed on the same loci of respective autosomes in both sexes. Southern hybridization analysis revealed an additional band including 17S rDNA exclusively on EcoRI digested female genomic DNA supporting the existence of an additional 17S rDNA locus on the X chromosome.     

Search for the sex-determining switch in monotremes: Mapping WT1 , SF1 , LHX1 , LHX2 , FGF9 , WNT4 , RSPO1 and GATA4 in platypus

The duck-billed platypus has five pairs of sex chromosomes, but there is no information about the primary sex-determining switch in this species. As there is no apparent SRY orthologue in platypus, another gene must acquire the function of a key regulator of the gonadal male or female fate. SOX9 was ruled out from being this key regulator as it maps to an autosome in platypus. To check whether other genes in mammalian gonadogenesis could be the primary switch in monotremes, we have mapped a number of candidates in platypus. We report here the autosomal location of WT1, SF1, LHX1, LHX9, FGF9, WNT4 and RSPO1 in platypus, thus excluding these from being key regulators of sex determination in this species. We found that GATA4 maps to sex chromosomes Y₁ and X₂; however, it lies in the pairing region shown by chromosome painting to be homologous, so is unlikely to be either male-specific or differentially dosed in male and female.     

Activity of the En/Spm-like transposons in meiosis as a base for chromosome repatterning in a small, isolated, peripheral population of Aegilops speltoides Tausch.

Chromosomal repatterning is considered to be one of the main mechanisms for plant genome evolution. Here, we report the first cytogenetic evidence for the involvement of En/Spm transposons in ongoing chromosomal repatterning leading to the rise of new fertile genomic forms in a small, isolated, peripheral plant population. Cytogenetical screening of original individual plants of Aegilops speltoides Tausch. with different phenotypes revealed a wide spectrum of chromosomal abnormalities including extra chromosomes, chromosomal rearrangements, and variability in chromosomal position/number of 45S and 5S rDNA sites. Cytogenetic analysis of the dynamics of En/Spm transposons in meiosis indicates that: (i) this type of transposon is active during male gametogenesis; (ii) separately or in conjunction with rDNA they form clusters in the hot spots of large chromosomal rearrangements; (iii) appearance of at least part of the mobile rDNA sites in genome of Ae. speltoides are connected with meiotic activity of En/Spm transposons. PCR screening for the site-selected transposon insertions confirm the presence of combined fragments that consist partly of the sequence of En/Spm transposon and partly of 5S rDNA sequence.     

Chromosomal Location and Nucleotide Sequences of 5S Ribosomal DNA of Two Cyprinid Species (Osteichthyes, Pisces)

5S ribosomal DNAs (rDNAs) from two cyprinid species, Acheilognathus tabira subsp. 1 and Cyprinus carpio, were isolated and sequenced. Tandemly arranged rDNAs were 179 bp in A. tabira and 204 bp in C. carpio. The non-transcribed spacer region elucidates the size difference of 5S rDNA between the two species. Fluorescence in-situ hybridization (FISH) localized 5S rDNAs to the short arms of two pairs of chromosomes in A. tabira and two to four pairs in C. carpio. Subsequent analysis demonstrated NORs in one pair of chromosomes in both species. Both the NOR and 5S rDNA are carried by a chromosome pair in A. tabira, but they are located on different chromosomes separately in C. carpio. Karyotype evolution by tetraploidy seems complex in cyprinid species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Ullrich-Turner phenotype with unusual manifestation in a patient with mosaicism 45,X/47,XX,+18

We report on a girl with Ullrich-Turner phenotype and 45,X/47,XX,+18 chromosomal mosaicism. Only two other patients with similar mosaicism have been reported, both girls with XY sex chromosome constitution. The face of the patient was highly asymmetric, the right side being almost normal, the left showing a typical Ullrich-Turner syndrome appearance. This clinical impression was strengthened by photographic doubling of both hemifaces. The patient had normal intelligence and did not show any stigmata of trisomy 18. © 1996 Wiley-Liss, Inc.     

Cell line segregation in a 45,X/46,XY mosaic child with asymmetric leg growth

Clinical evaluation of a 13 1/2-year-old male revealed a 4.4-cm leg length discrepancy and a small penis with a normal endocrine evaluation. Cytogenetic analysis of peripheral blood lymphocytes and skin fibroblasts derived from the back showed 45,X/46,XY mosaicism with similar percentages of 45,X cells, 36% and 30% respectively. However, two separate skin fibroblast cultures derived from the thigh and calf of the short (right) leg showed significant lack of Y-bearing cells with 100% and 80% 45,X, respectively. In contrast, skin biopsies of the thigh and calf of the normal (left) leg both showed 100% 46,XY. Similar evidence for differences in the percentages of Y-bearing cells in the left versus right leg fibroblast cultures was obtained using densitometric scanning of dot blots following DNA hybridization with a Y-specific probe at the DYZ4 locus. Asymmetric limb growth in cases of X/XY lymphocyte mosaicism warrants further cytogenetic investigation to substantiate possible genotype-phenotype correlations which may help uncover the fundamental growth deficiency in Turner syndrome.     

Organization of ribosomal RNA genes in Alternaria alternate Japanese pear pathotype,a host-selective AK-toxin-producing fungus

Summary DNA encoding ribosomal RNA (rRNA) of Alternaria alternata Japanese pear pathotype has been cloned in , replacement vector, , Fix. Restriction endonuclease mapping and Southern hybridization with the 18S and 28S rRNAs of Saccharomyces cerevisiae revealed the A. alternata rDNA to be tandemly repeating 8.15-kilobase pair unit. The restriction fragments of the unit were then subcloned in the plasmid vector Bluescribe M13- and partially sequenced. The determined sequences were compared with previously reported sequences of S. cerevisiae rRNAs and their genes. The locations of DNA sequences encoding the 5.8S, 18S, and 28S rRNAs were determined by homology search using reported sequences. The complete DNA sequence for 5.8S rRNA of the fungus was found to be highly conserved at more than 90 % homology in the fungi analyzed. However, sequence diversities were observed in limited regions involved in a helix structure, the helix (e), found at position 116–137.Deceased     

Differentiation and the Polymorphic Nature of the Y Chromosomes Revealed by Repetitive Sequences in the Dioecious Plant, Rumex Acetosa

The dioecious plant Rumex acetosa has a multiple sex chromosome system: females are 2n = XX + 12, males are 2n = XY₁Y₂ + 12, and the two Y chromosomes are heterochromatic. A DNA sequence abounded in the male genome was isolated and analyzed. The sequence (RAE180) was a 180-bp-long tandemly arranged repetitive sequence, distributed in chromosomes Y₁ and Y₂, and two pairs of autosomes. Both Y chromosomes contained large amounts of RAE180 and the sequence formed many DAPI bands, while, on the two pairs of autosomes, RAE180 did not form DAPI bands. The internal structure and morphological changes of the Y chromosomes were analyzed by FISH, using RAE180 and the Y-chromosome-specific sequence RAYSI as probes. The pattern of the FISH signals caused by the accumulation of RAE180 and RAYSI suggested the structural change in the Y chromosomes during the process of sex chromosome evolution, and the morphological change in the Y chromosomes was explained by reciprocal translocation and inversion. This revised version was published online in August 2006 with corrections to the Cover Date.     

Novel homozygous mutations in Desert hedgehog gene in patients with 46,XY complete gonadal dysgenesis and prediction of its structural and functional implications by computational methods

Male to female sex reversal in patients with 46,XY karyotype results from the failure of development of testis which may be due to mutations in the SRY gene. Only 10–15% of cases of 46,XY gonadal dysgenesis are accounted for by different types of mutations in the SRY gene. Hence, majority of such patients may have mutations in other genes involved in the testicular differentiation pathway. Besides SRY, other autosomal and X-linked genes are also involved in sexual development during embryogenesis. We describe here the first report from India wherein, two cases of 46,XY complete gonadal dysgenesis that could be attributable to mutations in the Desert hedgehog (DHH) gene. The mutations found in these two patients were a homozygous deletion (c.271_273delGAG) that resulted in deletion of one amino acid (p.D90del) and a homozygous duplication (c.57–60dupAGCC) that resulted in premature termination resulting in non-functional DHH protein. The structure–function implications of the p.D90del mutation were predicted using computational tools. Structural studies on the p.D90del mutant revealed that the mutation could seriously perturb the interaction of DHH with its binding partners. This is the second report in literature showing homozygous mutation in cases with 46,XY complete gonadal dysgenesis.     

Tetrasomy 5p mosaicism in a boy with delayed growth,hypotonia, minor anomalies,and an additional isochromosome 5p [46,XY/47,XY, + i(5p)]

We describe a 1-year-old boy with a rare de novo 46,XY/47,XY,+ i(5p) mosaicism (ratios 28/3 in peripheral blood lymphocytes and 2/12 in skin fibroblasts). The boy, born after a pregnancy of 34 weeks, had lung hypoplasia, persistent hypotonia, and postnatal growth failure. Craniofacial anomalies were also present. His clinical manifestations correspond to those described in trisomy 5p patients. Prenatal diagnosis on maternal age indication had shown normal male chromosomes in 16 cells in the short term culture of a chorionic villus sampling. Retrospectively, 1 out of 217 cells in this culture showed the i(5p). Several mechanisms could have resulted in the formation of this 46/47, + i(5p) mosaic. Postzygotic local incorrect ligation during chromatid replication, followed by a second replication offers an attractive model on theoretical grounds since it needs only one step to explain both isochromosome formation and mosaicism. Differences between the various tissues in selection pressure on cells with the isochromosome might explain the different ratios of mosaicism found. © 1993 Wiley-Liss, Inc.     

Karyological and genetic variation in Middle Eastern lacertid lizards, Lacerta laevis and the Lacerta kulzeri complex: a case of chromosomal allopatric speciation

Karyological (standard and C, Ag-NOR and Alu-I banding methods) and mtDNA analyses (cytochrome b and 12S rRNA) were conducted on specimens from eight allopatric populations of the Lacerta kulzeri complex. Parallel analyses were performed for comparison on Lacerta laevis specimens. Karyological and molecular studies support the morphological and ethological evidence indicating the specific separation between Lacerta laevis and Lacerta kulzeri In the Lacerta kulzeri complex, chromosomal analysis substantiated an interpopulation differentiation roughly along a north–south trend, mainly regarding the sex chromosome morphology and heterochromatin.The cytochrome b and 12S rRNA gene analyses showed minor genetic differences that were considerably smaller than those commonly found in genetically isolated populations. The L. kulzeri populations from Barouk, Druze and Hermon show a mean genetic distance that, in other saurians, characterises subspecies.The conditions found in L. laevis and L. kulzeri are reminiscent of King's model of chromosomal primary allopatry and support the hypothesis that in these lacertid lizards chromosome variations can become fixed before the accumulation of the genetic mutations.     

A non-sex chromosome marker in a patient with an atypical Ullrich-Turner phenotype and mosaicism of 46,X,mar/46,XX

The absence of a sex chromosome in conjunction with the presence of a marker chromosome generally implicates a sex chromosome origin for such marker chromosomes. These types of findings are frequently associated with Ullrich-Turner syndrome. We report a patient that presented with an atypical Ullrich-Turner phenotype and a cytogenetic mosaicism of 46,X,mar/46,XX. The marker chromosome was derived from chromosome 20, not from the X or Y chromosome. The patient's clinical features are described and discussed relative to the cytogenetic findings. This case further demonstrates the necessity of marker chromosome identification for accurate phenotype-karyotype correlation.     

Accumulation of rare sex chromosome rearrangements in the African pygmy mouse, Mus (Nannomys) minutoides: a whole-arm reciprocal translocation (WART) involving an X-autosome fusion

Although sex chromosomes are generally the most conserved elements of the mammalian karyotype, those of African pygmy mice show three extraordinary deviations from the norm: (a) asynaptic sex chromosomes, (b) multiple sex–autosome fusions, and (c) modifications of sex determination in some populations/species. In this study we identified, in two sex-reversed females of Mus (Nannomys) minutoides, a fourth rare sex chromosome change: a spontaneous whole-arm reciprocal translocation (WART) between an autosomal Robertsonian pair Rb(13.16) and the sex–autosome fusion Rb(X.1). This represents one of the very few reported cases of WARTs in natura within mammals, and is the first one to involve sex chromosomes. Hence, this finding offers new insights into the mechanisms of chromosomal differentiation in African pygmy mice, as WARTs may have contributed to the extensive diversity not only of autosomal Robertsonian fusions, but also of sex–autosome translocations. More widely, these results provide additional support to previous studies on the house mouse and the common shrew which indirectly inferred the role of WARTs in their karyotypic evolution, and may even help to understand how the fascinating 10 sex chromosome chain of the platypus might have evolved. This accumulation of rare sex chromosome changes in single specimens is, to our knowledge, exceptional among mammals.