bluuebunny/crossref_metadata_2025 huggingface dataset:
Meiotic behavior of rye heterozygous for ditelocentric substitutions
Authors: Benavente E. , Santos J.L.
Published: May 2008

Meiotic behavior has been investigated in rye plants carrying a heterozygous ditelocentric substitution for chromosomes 1R and 3R. Such a chromosomal rearrangement replaces a biarmed chromosome with two telocentrics, a situation karyotypically comparable to a Robertsonian translocation. As in Robertsonian translocation heterozygotes, delayed pairing characterizes the centromeric region in the rye pachytene heteromorphic trivalents. In addition, telocentric axes appear to be frequently engaged in nonhomologous synapsis (e.g., fold-back loops and two-by-two heterosynapsis) at their centromeric ends. Unlike those in Robertsonian translocation heterozygotes, the synaptic abnormalities detected here cannot be due to lack of partner homology. Rather, the chromosomal rearrangement itself appears to interfere with normal synaptonemal complex (SC) progression. The synaptic irregularities, which often result in complex pachytene multivalents, seem to have no effect on chiasma formation, since the rearranged pairs behave properly at metaphase I. Finally, differences in both SC length and progression patterns between plants and animals could explain the trans configurations of telocentric centromeres in plants, whereas only cis configurations of acrocentric kinetochores have been described in animals.
Localization of the human cystatin D gene (CST5) to chromosome 20p11.21 by in situ hybridization
Authors: Freije J.P. , Pendás A.M. , Velasco G. , Roca A. , Abrahamson M. , López-Otín C.
Published: May 2008

The gene coding for cystatin D (CST5), a cysteine proteinase inhibitor, was mapped by fluorescent in situ hybridization to human chromosome 20p11.21. This assignment, together with previous data on mapping of members of the cystatin gene family, indicates that cystatin family II genes are all clustered on the short arm of chromosome 20, whereas cystatin family I and III genes are located on the long arm of chromosome 3.
Localization of the human oncostatin M gene (OSM) to chromosome 22q12, distal to the Ewing’s sarcoma breakpoint
Authors: Giovannini M. , Selleri L. , Hermanson G.G. , Evans G.A.
Published: May 2008

Using fluorescence in situ hybridization, a cosmid clone containing the gene for oncostatin M (OSM) was mapped to human chromosome 22q12, placing the OSM gene in the same chromosome band as the leukemia-inhibitory factor gene (LIF). The location of the OSM gene was determined relative to the t(11;22)(q24;q12) of Ewing’s sarcoma and found to be distal to the translocation breakpoint on chromosome 22. Analysis of physical distances by pulsed-field gel electrophoresis demonstrated further that the two genes lie within 500 kb of each other.
Clonal expansion of cells with trisomy of chromosomes 12 and X in an EBV-transformed lymphoblastoid cell line and establishment of a tumorigenic monoclonal cell line (48, XX,+X,+12)
Authors: Risin S. , Fujimaki T. , Mestriner C.A. , Brown N.M. , Hopwood V.L. , Fidler I.J. , Pathak S.
Published: May 2008

Clonal expansion of cells with a certain abnormal chromosome constitution has been demonstrated in EBV-transformed lymphoblastoid cell lines, and a new monoclonal tumorigenic cell line (48, XX,+X,+12) has been established. Based on these data and our previous hypothesis that predisposition to tumor development could be associated with inherited susceptibility of certain chromosomes and chromosome regions to breakage, we emphasize that cytogenetic analysis of immortalized cell populations established from normal individuals and cancer patients might be of great importance for predicting development of different primary and secondary malignancies.
An irradiation-reduced hybrid panel for fine-structure mapping of the Xq28 region in the human genome
Authors: Peterlin B. , Smahi A. , Holvoet-Vermaut L. , Heitz D. , Dahl N. , Hors-Cayla M.C.
Published: May 2008

Irradiation-reduced somatic cell hybrids containing fragments of the human X chromosome were constructed. Analysis of 16 hybrids that retained the Xq28 region with 12 Xq28-specific markers identified at least six different breakpoints, supporting the order cen-DXS304-DXS374-(DXS33, DXS134, DXS52, DXS15)-RCP-(DXS254, G6PD, F8C)-(DXS115, DXYS64)-qter. The generated panel of hybrids provides a useful tool for fine mapping of probes in the Xq28 region.
A balanced autosomal reciprocal translocation in an azoospermic bull
Authors: Ansari H.A. , Jung H.R. , Hediger R. , Fries R. , König H. , Stranzinger G.
Published: May 2008

This appears to be the first reported case of a bull with a balanced autosomal reciprocal translocation associated with azoospermia. The analysis includes somatic chromosome banding, conventional meiotic analysis, and electron microscopy of synaptonemal complexes (SCs). The karyotype of the bull was found to be 60, XY, rcp(8;13)(ql l;q24). Electron microscopy of SCs in microspread pachytene spermatocytes revealed a high incidence of terminal asynapsis of the smallest arm of the quadrivalent. Most quadrivalents with such asynapsis and few with nonhomologous synapsis showed associations with the XY sex bivalent, leading to complete meiotic arrest at late pachynema. Except for one diakinetic cell, no diplotene and subsequent stages were encountered in air-dried meiotic preparations. The presence of degenerating primary spermatocytes in SC preparations, as well as in testicular sections, and the absence of spermatozoa in ejaculates confirm the chromosomally derived male sterility of the bull. X-chromosome reactivation, evidenced by the cytomorphological reversal of associated sex bivalents, appeared to be the initial step in the degeneration of spermatocytes. Consequently, the formation of a separate, fully developed XY body, which was previously demonstrated on the periphery of spermatocyte nuclei in fertile bulls, could not be attained in this case. Extensive end-to-end association of autosomal bivalents in meiotically arrested, as well as degenerating, spermatocytes was a consistent and unique observation of this study. Such associations may lead to enhanced reactivation of the X chromosome.
Synaptonemal complex analysis of a reciprocal translocation, rcp(20;24) (q17;q25), in a subfertile bull
Authors: Villagómez D.A.F. , Andersson M. , Gustavsson I. , Plöen L.
Published: May 2008

A reciprocal translocation was identified in a subfertile artificial insemination bull. Somatic chromosome investigation of G-banded metaphases demonstrated a 60, XY, rcp(20;24)(q17;q25) karyotype for the carrier. Synaptonemal complex analysis of the translocation by electron microscopy revealed an irregular pairing behavior of the chromosome axes involved, which resulted in a variety of configurations at pachytene. Not only was the expected quadrivalent configuration present, but also a trivalent plus univalent and two heteromorphic bivalents. Most common was an incompletely or completely paired quadrivalent configuration, which was non-homologously paired. XY association with the multivalent was seen only rarely. Histological analysis of testicular tissue showed meiotic arrest in some tubules. However, the semen picture was normal.
Comparative analyses of heterochromatin in Microtus: sequence heterogeneity and localized expansion and contraction of satellite DNA arrays
Authors: Modi W.S.
Published: May 2008

Southern blotting, C-banding, base-specific fluorochrome staining, and fluorescence in situ hybridization were used to analyze the constitutive heterochromatin in eight species and subspecies of arvicolid rodents (genus Microtus). Autosomal centromeric regions portrayed considerable variability between species in the amount of C-band-positive material present; e.g., M. chrotorrhinus showed relatively little, whereas M. cabrerae exhibited extensive centromeric staining. Autoso-mal interstitial C-bands were noted in M. guentheri and two subspecies of M. ochrogaster. All Y chromosomes examined were predominately or completely heterochromatic, as were substantial portions of the giant X chromosomes in three species (M. agrestis, M. cabrerae, and M. chrotorrhinus). Hoechst 33258 staining (with its affinity for AT binding sites) showed bright fluorescence on the heterochromatin of the sex chromosomes of M. agrestis and moderate fluorescence on those of M. cabrerae and M. chrotorrhinus; however, only the heterochromatin of M. cabrerae and M. chrotorrhinus hybridized with an AT-rich satellite DNA probe (MSAT-160) isolated from M. chrotorrhinus. Hoechst 33258-bright autosomal centromeres of M. arvalis and M. cabrerae also hybridized to the probe, whereas the Hoechst 33258-bright Y chromosomes of M. arvalis and M. guentheri did not. Two pairs of autosomes in M. guentheri are comprised of six distinct regions, based upon C-banding, Hoechst 33258 staining, chromomycin A3/distamy-cin A staining, and in situ hybridization. The centromeric regions of acrocentric autosomes known to retain conserved G-banding patterns may exhibit variable hybridization intensity when different species or subspecies are compared. M. ochrogaster portrays considerable intersubspecific variability in the size and location of autosomal telomeric and interstitial C-bands that are also sites of hybridization. These latter two findings illustrate that dramatic differences in copy number of the tandem satellite array can exist at homologous chromosomal positions both within and between species.
New markers, D16FC1 and Tp12, differentiate between rat chromosomes 16 and 17
Authors: Yeung R.S. , Taguchi T. , Patriotis C. , Makris A. , Tsichlis P.N. , Levan K.K. , Levan G. , Tartof K. , Hino O. , Knudson A.G. , Testa J.R.
Published: May 2008

Problems in differentiating rat chromosomes 16 and 17 cytogenetically can be resolved with unique probes mapped to these chromosomes. Using somatic cell hybridization and nonisotopic in situ hybridization, probes D16FC1 and Tp12 were localized to 16p16→p15 and 17q12.1→q12.2, respectively. The locations of these probes can serve as reference points to facilitate mapping of future probes to rat chromosomes 16 and 17.
Gene amplification and chromosome rearrangements: a study of a single cell lineage selected for amplification and deamplification of the UMP synthase gene
Authors: Qumsiyeh M.B. , Goorha S. , Suttle D.P.
Published: May 2008

The UMP synthase gene is stably amplified in Chinese hamster lung cells selected for resistance to pyrazofurin (PF) and 6-azauridine (6AUR), inhibitors of the decarboxylase activity of the bifunctional UMP synthase enzyme. The amplified DNA is located intrachromosomally as expanded chromosomal regions (ECRs). Growth of these cells in 5-fluorouracil enables rapid selection of cells that have undergone deamplification and consequently lost resistance to PF + 6AUR. Detailed cytogenetic analyses and fluorescence in situ hybridization on three consecutive amplification-deamplification cycles in descendants of the same cloned cell showed a unique position and structure for the ECR. In the first cycle of amplification, the ECR forms a homogeneously staining region on a small marker chromosome (M3). In the second cycle of amplification, a chromosomal break was noted at the site of the endogenous UMP synthase gene on another derivative chromosome, M2, with amplification resulting in an abnormally banded region on a third marker (M3). The third cycle of amplification produced a cell line with an ECR on the distal portion of M2. This ECR was unstable, showing variations in size as well as translocations and other chromosome rearrangements. Our data, taken in its entirety, suggest a relationship between amplification as an ECR and chromosome rearrangements in Chinese hamster cells.
Regional sublocalization of the human thyroid peroxidase gene (TPO) by tritium and fluorescence in situ hybridization to chromosome 2p25→p24
Authors: Barnett P.S. , Jones T.A. , McGregor A.M. , Banga J.P. , Sheer D.
Published: May 2008

Thyroid peroxidase (TPO), the enzyme responsible for thyroid hormone synthesis, is a key autoantigen in autoimmune thyroid disease. Based on the results of a study using both isotopic and nonisotopic in situ hybridization, we assign the gene coding for human TPO to chromosome region 2p25→p24.
Localization of the human vitamin D 24-hydroxylase gene (CYP24) to chromosome 20q13.2→q13.3
Authors: Hahn C.N. , Baker E. , Laslo P. , May B.K. , Omdahl J.L. , Sutherland G.R.
Published: May 2008

The gene encoding human vitamin D 24-hydroxylase (P45024) has been localized by fluorescence in situ hybridization to 20q13.2→q13.3.
In humans all U3 genes map to chromosome 17p12→p11, but in mouse the U3A and U3B genes are located on different chromosomes
Authors: Mazan S. , Mattéi M.-G. , Roeckel N. , Qu L.-H. , Bachellerie J.-P.
Published: May 2008

U3 small nucleolar RNA, which participates in eukaryotic rRNA processing, is encoded by a small multigene family in mammals. In humans, the four to six gene copies code for an identical U3 RNA molecule; rodents, however, have two variant forms of these genes, U3A and U3B. We show that all U3 genes in humans map to a single chromosomal locus, 17p12→p11, which corresponds exactly to the region of mouse Chromosome 11 where the four U3B genes are clustered. By contrast, in mouse the unique U3A gene copy is not linked to the U3B gene cluster but maps to another chromosome (the B2-B4 region of Chromosome 10).
Newly acquired peri-telomeric heterochromatin in a transgenic mouse lineage
Authors: Simon D. , Knowles B.B.
Published: May 2008

Karyotypes of an embryonic stem cell line and normal bone marrow and peripheral blood cells from a transgenic mouse lineage, strain 83, exhibit a novel region of constitutive heterochromatin in the peri-telomeric region of chromosome 3. This heterochromatic region co-localizes with the integration site of the transgene, an 11-Mb tandem insertion of a plasmid containing human β-globin gene sequences, and is inherited in Mendelian fashion. The significance of this finding is discussed, and a nomenclature for such newly acquired heterochromatic regions is suggested.
Novel RFLPs at protooncogene and cancer-related gene loci on mouse chromosomes
Authors: Santos J. , Pellicer A.
Published: May 2008

DNA probes for the NRAS, HRAS, KRAS2, LCK, RAF1, MET, MYCL1, MYCN, MYB, ERBB2, FOS, CSFIR, and SRC protooncogene loci; the retinoblastoma gene locus (RBI); the tumor virus integration sites INT2, PVT1, and MLVI2; and the locus of the tumor-specific antigen T1A were used to screen mouse genomic DNAs from RF/J, CAST/Ei, MOLF/Ei, Mus musculus musculus, M. m. poschiavinus, and M. spretus. Polymorphic DNA fragments for the 18 DNA probes have been identified using Southern blot hybridization and restriction fragment length polymorphism (RFLP) analysis.
Characterization of the karyotype of the tench (Tinca tinca L.) and analysis of its chromosomal heterochromatic regions by C-banding, Ag-staining, and restriction endonuclease banding
Authors: Padilla J.A. , Fernández-García J.L. , Rabasco A. , Martínez-Trancón M. , Rodriguez de Ledesma I. , Pérez-Regadera J.J.
Published: May 2008

Beginning with a description of the conventional Giemsa-stained karyotype of the tench (Tinea tinea L.), the structure and variability of the chromosomal heterochromatic regions in this cyprinid species were analyzed by means of C-, silver-, and restriction endonuclease banding. Silver staining revealed active nucleolus organizer regions (NORs) on the secondary constriction of chromosome pair 3. Constitutive heterochromatin was associated with NOR regions detected by C-banding. Restriction endonuclease digestion with AluI, TaqI, and HaeIII induced specific banding patterns that allowed identification of homologous chromosome pairs and revealed features about the sequence composition of several chromosomal heterochromatic regions and of the NOR-associated heterochromatin.
A simple G-banding technique following nonradioactive in situ hybridization
Authors: Duncan L.M. , Dietzsch E. , Rudduck C. , Garson O.M.
Published: May 2008

A simple G-banding protocol has been found to be compatible with a nonisotopic in situ hybridization technique utilizing a biotin-streptavidin-polyalkaline phosphatase complex. The chromosomes are banded following detection of the hybridization signal, the procedure requires no pretreatment of cultures or special optical equipment, and it produces permanent preparations.
Detection of a new submicroscopic Norrie disease deletion interval with a novel DNA probe isolated by differential Alu PCR fingerprint cloning
Authors: Bergen A.A.B. , Wapenaar M.C. , Schuurman E.J.M. , Diergaarde P.J. , Lerach H. , Monaco A.P. , Bakker E. , Bleeker-Wagemakers E.M. , van Ommen G.J.B.
Published: May 2008

Differential Alu PCR fingerprint cloning was used to isolate a DNA probe from the Xp 11.4→p11.21 region of the human X chromosome. This novel sequence, cpXr318 (DXS742), detects a new submicroscopic deletion interval at the Norrie disease locus (NDP). Combining our data with the consensus genetic map of the proximal short arm of the X chromosome, we propose the physical order Xcen – DXS14 – DXS255 – (DXS426, TIMP) – (DXS742 – ([MAOB – MAOA -DXS7], NDP) – DXS77 – DXS228) – DXS209 – DXS148 -DXS196 – Xpter. The cpXr318 probe and a subclone from a cosmid corresponding to the DXS7 locus were converted into sequence-tagged sites. Finally, DXS742, DXS7, DXS77, and MAOA were integrated into a physical map spanning the Norrie disease locus.
Linkage analysis of the monoamine A and B genes using newly-defined polymorphisms
Authors: Harris B.D. , Kurth J.H. , Barnes R.I. , Bowcock A.M. , Kurth M.C.
Published: May 2008

The monoamine oxidase A (MAOA) and B (MAOB) genes have been localized to chromosome Xp11.3. Recently-defined polymorphisms and linkage analysis have shown tight linkage between MAOA and MAOB with a distance of approximately 2.7 cM between them.
Genome size and qualitative and quantitative characteristics of C-heterochromatic DNA in Eulemur species and in a viable hybrid
Authors: Ronchetti E. , Crovella S. , Rumpler Y. , Pellicciari C. , Manfredi Romanini M.G.
Published: May 2008

The amounts of nuclear DNA and the AT and GC content of four Eulemur (Prosimii, Lemuridae) species and of an E. coronatus × E. macaco hybrid were measured by flow cytometry in peripheral blood leukocytes, following propidium iodide, Hoechst 33258, and mithramycin staining. Hoechst 33258 and mithramycin were also used to evaluate the base composition of genomic DNA in the chromosomes. The amount of DNA resisting C-banding pretreatment (C-heterochromatic DNA) was measured in metaphase chromosomes by static fiuorometry. The genome of E. coronatus was significantly larger than the genomes of all other species examined, due to a higher content of pericentromeric, mainly GC-rich, heterochromatic DNA. The restriction banding patterns produced by BamRl digestion and ethidium bromide staining on extracted DNA were studied in the hybrid and its parental species (E. coronatus and E. macaco). The restriction banding pattern of the sole E. coronatus individual showed two bands which were repeated in the restriction banding pattern of the hybrid. The qualitative and quantitative differences of C-heterochromatic DNA in E. coronatus confirm the “splitting” processes and the phylogenetic relationships in the genus Eulemur suggested by Jung et al. (1992) on the basis of the restriction banding patterns produced by endonuclease digestion.
Analysis of chiasma frequency and first meiotic segregation in a human male reciprocal translocation heterozygote, t(1;11) (p36.3;q13.1), using fluorescence in situ hybridisation
Authors: Goldman A.S.H. , Hultén M.A.
Published: May 2008

In this study we have used a testicular biopsy from a human male with a 46, XY, t(1;11)(p36.3;q13.1) karyotype. Fluorescence in situ hybridisation with whole chromosome libraries and paracentromeric probes were applied to identify normal and derived chromosomes 1 and 11 in both first metaphase (MI) and second metaphase (MII) cells. The chiasma frequency distribution was established in the quadrivalent. A large proportion of MI cells was found to have at least one interstitial chiasma, resulting at MII in dimorphic chromosomes bearing one normal and one translocated chromatid. Alternate, adjacent I, adjacent II, and 3:1 products were all identified at MII. More than half of the cells analysed could not be assigned to a single segregation category because of the presence of interstitial chiasmata. Such MII cells could have arisen from either alternate or adjacent I segregation. We also calculated the proportion of sperm expected to be normal, balanced, and unbalanced. The latter data are in agreement with the results reported by Spriggs et al. (1992), who karyotyped sperm from the same individual.
Localization of the genes encoding the catalytic subunits of protein phosphatase 2A to human chromosome bands 5q23→q31 and 8p12→p11.2, respectively
Authors: Jones T.A. , Barker H.M. , da Cruz e Silva E.F. , Mayer-Jaekel R.E. , Hemmings B.A. , Spurr N.K. , Sheer D. , Cohen P.T.W.
Published: May 2008

The gene for the α isoform of the catalytic subunit of human protein phosphatase 2A (PPP2CA) was localized to chromosome 5 using somatic cell hybrids, and then more finely mapped to chromosome region 5q23→q31 by in situ hybridization using a tritiated cDNA probe. The gene for the β isoform of the catalytic subunit of this enzyme (PPP2CB) was mapped by the polymerase chain reaction to human chromosome 8 using somatic cell hybrids. Fluorescence in situ hybridization was then used to localize the PPP2CB gene to 8p12→p11.2, using a mixture of three genomic probes that ranged from 3.5 to 8 kb in size. Finally, Southern blot analysis of somatic cell hybrid DNA suggested that a PPP2CB catalytic subunit pseudogene (PPP2CBP) is on chromosome 16.
The probe BMS1271 identifies a new polymorphic locus (D3S1207) and maps to 3p26-pter
Authors: Heppell-Parton A.C. , Daly M.C. , Drabkin H.A. , Rabbitts P.H.
Published: May 2008

We report here the characterization of a new polymorphic locus (D3S1207), shown by fluorescence in situ hybridization to map to 3p26-pter. The distal region of chromosome 3 has been implicated in both von Hippel-Lindau and 3p deletion syndrome, lung, uterine, breast, testicular, and ovarian cancers, renal cell and nasopharyngeal carcinomas, mesotheliomas, and various haematological malignancies, yet relatively few polymorphic marker probes are well mapped within this region. The identification of this new probe will therefore be particularly useful for the study of these diseases.
Multiple and sensitive fluorescence in situ hybridization with rhodamine-, fluorescein-, and coumarin-labeled DNAs
Authors: Wiegant J. , Wiesmeijer C.C. , Hoovers J.M.N. , Schuuring E. , d’Azzo A. , Vrolijk J. , Tanke H.J. , Raap A.K.
Published: May 2008

We have tested the use of several newly developed red, green, and blue fluorescent dUTPs in direct, multiple, and sensitive fluorescence in situ hybridization procedures. Among the ones tested, the tetramethylrhodamine-dUTP proved to give the best sensitivity; using conventional epifiuorescence microscopy, cosmids could be visualized with a hybridization efficiency of 90%. Fluorescein-dUTP permitted visual cosmid detection with 50% efficiency, and, to the human eye, the green signals appeared less bright. With a blue fluorescent coumarin-labeled dUTP, only highly repetitive target sequences could be visualized directly. Imaging with a cooled CCD (charged coupled device) camera for prolonged integration times permitted localization of small targets using probes labeled with red and green fluorochromes. Visualization of a 1.8-kb single copy sequence with a rhodamine-labeled cDNA proved feasible. The strategy of identifying hybridization sites with multiple colors was successfully applied for multiple hybridizations to repetitive and unique targets, using the red and green labels directly and the biotin label indirectly with one layer of avidin-coumarin.
Deleted and normal chromosome 10 homologs from a patient with Hirschsprung disease isolated in two cell hybrids through enrichment by immunomagnetic selection
Authors: Puliti A. , Covone A.E. , Bicocchi M.P. , Bolino A. , Lerone M. , Martucciello G. , Jasonni V. , Romeo G.
Published: May 2008

A cytogenetically detectable deletion, del(10) (q11.2→q21.2), was observed in a patient with total colonic aganglionosis with small bowel involvement (TCSA), a variant of Hirschsprung disease (HSCR). A similar deletion is present in another TCSA patient (S.M. Huson, personal communication). To reveal cytogenetically undetectable deletions of chromosome 10 in further patients, we developed a strategy for mapping chromosome 10 DNA markers with respect to the observed deletions. To this end, the two chromosome 10 homologs (deleted and normal) were segregated in two distinct somatic cell hybrids obtained after fusion of the patient’s fibroblasts with a Chinese hamster ovary cell line (YH21). Hybrid cells containing chromosome 10 were selected for the expression of the gene coding for the β subunit of the fibronectin receptor (FNRB), which maps to lOpl 1.2, using a monoclonal antibody against FNRB. Hybrid 185.0 contains the deleted chromosome, whereas hybrid 179.Q contains the nondeleted one. Southern blot and PCR analysis of DNA from these two hybrids mapped the markers RBP3H4, RET, D10S15, D10S5, D10S22, and D10S88 inside the deletion and D10S170, CDC2, EGR2, and D10S19 outside the deletion. MEN2A and MEN2B have recently been mapped within the centromeric region closely linked to RBP3 and D10S15 (which are located inside the deletion) and cosegregate with HSCR in at least two different pedigrees. Since HSCR, MEN2A, and MEN2B represent defects of neural crest cell development, we hypothesize that they originate from mutations in different genes clustered in the centromeric region of 1 Oq.
Rapid immunophenotypic characterization of chromosomally aberrant cells by the new FICTION method
Authors: Weber-Matthiesen K. , Deerberg J. , Müller-Hermelink A. , Schlegelberger B. , Grote W.
Published: May 2008

Recently, we have presented a new technique for immunophenotyping cells that have numerical chromosome aberrations. We referred to this method as “Fluorescence-Immunophenotyping and Interphase Cytogenetics as a Tool for Investigation of Neoplasms” (FICTION). We present here an advanced FICTION method with three-color staining and improved sensitivity.
Comparison of highly repeated DNA sequences in some Lemuridae and taxonomic implications
Authors: Montagnon D. , Crovella S. , Rumpler Y.
Published: May 2008

Highly repeated DNA sequences of Eulemur fulvus mayottensis, E. coronatus, Lemur catta, and Hapalemur griseus griseus have been identified and compared. Sequence analysis of highly repeated DNA fragments isolated from L. catta and Hapalemur showed a high percentage of similarity (nearly 95 %), as did fragments isolated from the two very close Eulemur species, whereas comparison of the DNA fragments isolated from the two Eulemur species and the L. catta/Hapalemur group showed a very low percentage (∼40%) of identity, as might be expected for distant species. These results confirm our previous data, obtained by Southern blot hybridization techniques on the same species, and strongly support the existence of a common trunk between L. catta and Hapalemur, but different from that leading to the Eulemur species.
Chromosomal evidence for a hybrid origin of diploid parthenogenetic females from the unisexual-bisexual Lepidodactylus lugubris complex (Reptilia, Gekkonidae)
Authors: Volobouev V. , Pasteur G. , Ineich I. , Dutrillaux B.
Published: May 2008

Chromosome banding analysis of seven diploid parthenogenetic females from Polynesian and Southeast Asian populations of the unisexual-bisexual Lepidodactylus lugubris complex (Reptilia, Gekkonidae) showed that the unisexual karyotype consists of two different haploid sets. Similar analysis of an L. lugubris male from a bisexually reproducing population revealed two identical chromosome complements, the banding pattern generally matching that of the females, although many chromosomes were obviously different. These observations suggest that the gonochoristic males belong to a taxon related to the parthenogenetic female taxon but are not a direct ancestor of the latter. The data also offer strong evidence for a hybrid origin of diploid parthenogenetic females in L. lugubris and suggest that a high degree of chromosomal heterogeneity may be the leading reason for the evolutionary selection of a modified meiotic process in this species, allowing production of unreduced oocytes and, consequently, of a unisexual mode of reproduction.
The OLFR1 gene encoding the HGMP07E putative olfactory receptor maps to the 17p13→p12 region of the human genome and reveals an MspI restriction fragment length polymorphism
Authors: Schurmans S. , Muscatelli F. , Miot F. , Mattei M.-G. , Vassart G. , Parmentier M.
Published: May 2008

Olfactory receptors are believed to be encoded by an extremely large subfamily of G protein-coupled receptors. A human gene (OLFRl) encoding a member of this family was cloned from a genomic library by cross-hybridization with a gene fragment obtained by the polymerase chain reaction. The nucleotide sequence of a 3.4-kb EcoRl fragment was determined, and the protein sequence was deduced from the single open reading frame. The gene was assigned by in situ hybridization of metaphase chromosomes to the 17p13→p12 region of the human genome, in proximity to the tumor-suppressor gene encoding p53. When used as a probe on Southern blots under moderately stringent conditions, it hybridizes to at least three closely related genes. A restriction fragment length polymorphism was detected after MspI digestion. Mendelian segregation of the gene was assessed in three CEPH families, and linkage analysis confirmed the localization of the locus.
Assignment of genes for interleukin-8 receptors (IL8R) A and B to human chromosome band 2q35
Authors: Lloyd A. , Modi W. , Sprenger H. , Cevario S. , Oppenheim J. , Kelvin D.
Published: May 2008

The human genes encoding the interleukin 8 receptors type A and B were assigned to chromosome 2 by polymerase chain reaction amplification and by Southern analysis of a panel of human × rodent somatic cell hybrid DNAs. The IL8R genes were further localized by in situ hybridization to band 2q35.
Sex chromatin in hair roots — 25 years later: fluorescence in situ hybridization of hair root cells for detection of numerical chromosome aberrations
Authors: Lampel S. , Steilen H. , Zang K.D. , Wullich B.
Published: May 2008

The applicability of fluorescence in situ DNA hybridization to hair root preparations for the detection of numerical chromosome anomalies is demonstrated. This simple method yields a high number of scorable nuclei, allowing rapid identification of epithelial cell chromosome composition. In clinical genetics, this technique may become useful in cases where other types of cells are difficult to obtain.
Isolation of new probes in the region of the Wilson disease locus, 13q14.2→q14.3
Authors: Bull P.C. , Barwell J.A. , Hannah H.T.-L. , Pautler S.E. , Higgins M.J. , Lalande M. , Cox DW.
Published: May 2008

A hybrid panel was used to refine the localizations of eight established markers useful for the diagnosis of Wilson disease. Two of the markers, D13S59 and D13S31 that map very close to the Wilson disease locus (WND) were localized to the region 13q14.2 → q14.3. We report the isolation of seven new probes from this region, using two different approaches. First, 16 clones from a chromosome 13-specific library were mapped using the hybrid panel. Three of the clones mapped to 13q14.2→q14.3. As a second approach, Alu element-mediated PCR (Alu-PCR) was used to generate clones from a hybrid (ICD) that contains the proximal half of chromosome 13 as the only human component. To select for those that potentially mapped within the region 13q14.2→q14.3, the clones were screened by differential hybridization using the labeled Alu-PCRproducts from a hybrid (KSF39) that is similar to ICD but has a deletion in the region 13q14.2→q14.3. The procedure was successful even though KSF39 contains five additional human chromosomes. Six independent clones were selected. Five of these were found to be nonrepetitive, and four were found to map correctly tol3q14.2→q14.3 when localized using the hybrid panel.
Localization of the gene encoding proteinase-3 (the Wegener’s granulomatosis autoantigen) to human chromosome band 19p13.3
Authors: Sturrock A.B. , Espinosa III R. , Hoidal J.R. , Le Beau M.M.
Published: May 2008

Proteinase-3 (PR3) is a polymorphonuclear leukocyte serine proteinase that is a potent inducer of emphysema in experimental models. PR3 is also the target antigen associated with Wegener’s granulomatosis. By analysis of a panel of somatic cell hybrids, the gene encoding PR3 was localized previously to chromosome 19; we have used fluorescence in situ hybridization to sublocalize this gene to 19p13.3.
Cytogenetic characterization of the AS cell line derived from the Atlantic salmon (Salmo salar L.)
Authors: Sánchez L. , Abuín M. , Amaro R.
Published: May 2008

The chromosome complement of the fish cell line AS, derived from the Atlantic salmon (Salmo sαlαr L., 2N = 58), was investigated by C-, Ag-NOR, restriction endonuclease, and BrdU-replication banding. The cell line has a modal chromosome number of 52 with a fixed number of chromosomal rearrangements. The variety of banding patterns obtained allowed us to identify three chromosome markers in the cell line. The identification of their possible origin and significance in karyotype evolution are discussed.
Linkage mapping detects two secondary microdeletions in cell hybrid HHW1064, used to isolate DNA probes from within 5q11.2→q13.3
Authors: Bernard L.E. , McPherson J.D. , Wasmuth J.J. , Kreklywich C.N. , Brzustowicz L.M. , Wood S.
Published: May 2008

The somatic cell hybrid HHW1064 contains a single human chromosome 5 with an interstitial deletion for 5q11.2→q13.3. Twenty human clones were isolated by Alu-PCRdifferential hybridization. Mapping these clones indicates that HHW1064 contains two additional secondary microdeletions.
Fine mapping of the human MHC class II region within chromosome band 6p21 and evaluation of probe ordering using interphase fluorescence in situ hybridization
Authors: Senger G. , Ragoussis J. , Trowsdale J. , Sheer D.
Published: May 2008

Eight previously well-characterized and mapped probes derived from the human major histocompatibility complex (MHC) class II region were used to investigate the advantages and limitations of fluorescence in situ hybridization (FISH) techniques for fine mapping. The class II region of the MHC was localized within subband 6p21.31 by in situ hybridization on metaphase chromosomes that were banded by immunofluorescence staining with an antibody against 5-bromodeoxyuridine (BrdU). Ordering of probes that were separated by up to 900 kb was achieved by simultaneous hybridization of two or three probes on interphase nuclei. Three-color FISH proved to be an excellent method for ordering of probes within distances of 200–1,000 kb. Under certain conditions, closer probes could be ordered by comparing measured distances between their hybridization signals in interphase nuclei. A linear correlation between measured interphase distances and kilobase distances was observed up to 500 kb. With increasing distances, the measurements become more inaccurate due to chromatin folding.
Assignment of the β subunit of the muscle nicotinic acetylcholine receptor gene (CHRNB1) to sheep chromosome 11
Authors: Pearce P.D. , Ansari H.A. , Maher D.W. , Malcolm A.A. , Wood N.J. , Phua S.H. , Broad T.E.
Published: May 2008

The chromosomal assignment of the gene for the β subunit of the sheep muscle nicotinic acetylcholine receptor (CHRNB1) was determined by isotopic in situ hybridization. CHRNB1 was mapped to sheep chromosome 11 with a hybridization peak at 11q14→q22. This result is further evidence that the chromosomal segment that comprises human chromosome 17 has been highly conserved during evolution.
A mouse stock with 38 chromosomes derived from the reciprocal translocation T(7;15)33Ad
Authors: Schriever-Schwemmer G. , Adler I.-D.
Published: May 2008

A reciprocal translocation, T(7;15)33Ad, with presumed breakpoints in bands 7A1 and 15F3 was induced in late spermatids by injecting male (102/E1 × C3H/E1)F! mice five times with acrylamide (50 mg/kg body weight). Outcrosses of the original semisterile T(7; 15) female generated three males monosomic for the short marker 715 [Ms(715)] among a total of 15 males. The Ms(715) males sired small litters and had reduced testes weights. From inter se matings of Ms(715) animals, nullisomic progeny for chromosome 715 were obtained and mated to produce a breeding stock of mice with 38 chromosomes. For comparison, mice carrying the reciprocal translocation T(4;8), with similarly located breakpoints, were also analyzed. Fluorescent in situ hybridization (FISH) with major and minor satellite DNA probes and a telomeric DNA probe was utilized. The observed FISH signals suggest that in chromosomes 7 and 8 the breaks occurred within the pericentric heterochromatic block immediately below the centromere and in chromosomes 15 and 4 at a point near the distal telomeres. The long markers 157 and 48 are tandem fusion chromosomes. The short markers 715 and 84 also showed all appropriate FISH signals for intact chromosomes. The loss of the small chromosome 715 was compatible with survival, suggesting that no essential genes are located on the small reciprocal translocation product. The development of this tandem fusion stock is described as a laboratory example of one possible step in karyotypic evolution.
Localization of the transcription factor SP1 gene to human chromosome 12q12→q13.2
Authors: Gaynor R.B. , Shieh B.-H. , Klisak I. , Sparkes R.S. , Lusis A.J.
Published: May 2008

The cellular transcription factor SP 1 binds to critical regulatory elements in a variety of cellular and viral promoters. The gene encoding the ∼100-kDa SP 1 protein contains zinc finger DNA-binding domains and glutamine-activation domains. Since SP1 is involved in the regulation of a variety of cellular genes, we wished to determine its chromosomal localization. Southern blot analysis of genomic DNA from a panel of mouse × human somatic cell hybrids indicated that SP1 was localized to chromosome 12. In situ hybridization allowed the localization of the gene encoding SP 1 to human chromosome 12q12→q13.2, with 12q13.1 being the most probable location.
Detection of a male-specific sequence in nonhuman primates through use of the polymerase chain reaction
Authors: Reitsma M.J. , Harrison M.R. , Pallavicini M.G.
Published: May 2008

Sex-specific DNA sequences are useful for detecting and monitoring chimerism in transplant recipients that had received sex-mismatched donor cells. Nonhuman primates are often used as experimental transplant models because of their evolutionary proximity, and the similarity of their physical characteristics, to those of humans. Unfortunately, DNA-based molecular detection strategies to monitor engraftment in sex-mismatched transplants in monkeys and baboons have not been available. We describe development of a polymerase chain reaction-based assay to detect a 174-bp male-specific sequence present in the rhesus monkey (Macaca mulatta) and olive baboon (Papio cynocephalus). The assay is sufficiently sensitive to allow detection of 10 male cells against a background of 104 female cells. Human sequence is not amplified under the described assay conditions. The amplified DNA sequence is 82% homologous to a sequence located near the testis-determining factor locus in the human genome, suggesting a high degree of evolutionary conservation in this region.
Mapping the intron-containing human hsp90α (HSPCAL4) gene to chromosome band 14q32
Authors: Vamvakopoulos N.C. , Griffin C.A. , Hawkins A.L. , Lee C. , Chrousos G.P. , Wang Jabs E.
Published: May 2008

The human heat-shock protein (hsp) 90 gene superfamily consists of two member forms, Α and Β. We have mapped the intron-containing human hsp90Α expressed gene (HSPCAL4) to chromosome band 14q32.
Genome distribution, chromosomal allocation, and organization of the major and minor satellite DNAs in 11 species and subspecies of the genus Mus
Authors: Garagna S. , Redi C.A. , Capanna E. , Andayani N. , Alfano R.M. , Doi P. , Viale G.
Published: May 2008

We compared the genome distribution, chromosomal allocation, and organization of the major and minor satellite DNAs (satDNAs) in 11 species and subspecies of the genus Mus. Southern blot analysis of the major and minor satDNAs showed similar fragment profiles in all 11 species, with the exception of M. cervicolor and M. cookii for the major satDNAs and M. caroli, M. cervicolor, and M. cookii for the minor satDNAs, where these sequences could not be detected by the probes used. In situ hybridization of the major and minor satDNA probes revealed chromosome-specific allocations of these sequences with quantitative species-specific patterns. Fluorometric analysis of the organization of the satellite sequences suggested that in the M. domesticus genome satDNA sequences are clustered in tandem repeats that are longer than those present in other Mus genomes. When compared with the other Mus genomes so far studied, the domesticus genome shows the highest quantity of satDNA sequences with a long-range organization of satDNA sequences.
Chromosomal anomalies and disturbance of transcriptional activity at the pachytene stage of meiosis: relationship to male sterility
Authors: Jaafar H. , Gabriel-Robez O. , Rumpler Y.
Published: May 2008

Morphological analysis of pachytene spermatocytes obtained from male mice carrying three chromosomal rearrangements – a Robertsonian translocation, Rb(X-2)2Ad; an autosomal reciprocal translocation, T(16;17)43H; and a tertiary trisomic, Ts(113)70H – demonstrated frequent association between the XY bivalent and the T43H and T70H translocation chromosomes. Quantitative autoradiographic data revealed that the normal transcriptional inactivity of the XY bivalent was not significantly disturbed, in contrast to that of the 16;17 quadrivalent and the extra 113 marker chromosome. These results are interpreted as an extension of the XY inactivation process to the associated autosomes and discussed in relation to male sterility.
Localization of the α-S2-casein gene (CASAS2) to the homoeologous cattle, sheep, and goat chromosomes 4 by in situ hybridization
Authors: Hayes H. , Petit E. , Bouniol C. , Popescu P.
Published: May 2008

The Α-S2-casein gene (CASAS2) has been mapped to the homoeologous cattle, sheep, and goat chromosomes 4 and to the long arm of a bovine chromosome translocation, t(4;8), using nonradioactive in situ hybridization and simultaneous fluorescent R-banding. The t(4;8) has been characterized by GTG-, GBG-, and RBG-banding and by silver staining of nucleolus organizer regions. These results confirm the previous (ISCNDA, 1989) localization of the casein gene group to chromosome 4 of cattle, sheep, and goats. We propose that the discrepancy between our results and earlier assignments of the casein gene group to chromosome 6 of cattle and sheep can be explained by the fact that the chromosome identified as No. 6 in the Reading Conference (1976) report was renamed chromosome 4 in the ISCNDA (1989) standardized karyotype of both species.
Flow cytometric detection of unbalanced ram spermatozoa from heterozygous 1;20 translocation carriers
Authors: Lewalski H. , Otto F.J. , Kranert T. , Wassmuth R.
Published: May 2008

Flow cytometric analysis of enzymatically decon-densed, DAPI-stained spermatozoa was performed to confirm the suspected production of unbalanced spermatozoa in heterozygous rams carrying a 1;20 translocation. High-precision flow cytometry (coefficient of variation, 0.6–0.8 %) with a PAS II flow cytometer depicted Y- and X-chromosome-bearing spermatozoa from three cytogenetically normal rams as two distinct peaks. The difference in DNA fluorescence intensity between the gonosomes averaged 4.8 %. Analysis of sperm samples from three heterozygous 1;20 translocation carriers yielded histograms with five peak distributions. The individual peaks were attributed to spermatozoa with a normal, balanced, and unbalanced chromosomal status. Peaks within Y- and X-spermatozoa populations were distributed in a ratio of 1:2:1 and were almost completely separated, with a coefficient of variation of 0.5–0.6%. Owing to the relative size of the translocated chromosomal segment (2.4% of the total DNA content, as determined from the flow cytometric data), histograms with five instead of the expected six peaks were observed.
Fine mapping of the human receptor-like protein tyrosine phosphatase gene (PTPRM) to 18p11.2 by fluorescence in situ hybridization
Authors: Suijkerbuijk R.F. , Gebbink M.F.G.B. , Moolenaar W.H. , Geurts van Kessel A.
Published: May 2008

The isolation of a cDNA encoding a receptor-like protein tyrosine phosphatase, termed RPTPµ, and the assignment of the human gene to chromosome 18 has recently been reported (Gebbink et al., 1991). Using a genomic clone and fluorescence in situ hybridization, we now show that the human gene, designated PTPRM (formerly PTPRLl; LeBeau and Geurts van Kessel, 1991), maps in band 18p11.2.
Mapping of 18 probes on human chromosome 18 using single- and double-color FISH
Authors: Muleris M. , Apiou F. , Olschwang S. , Thomas G. , Dutrillaux B.
Published: May 2008

Fluorescence in situ hybridization (FISH) techniques were applied to the mapping of 18 probes from chromosome 18, permitting seven new assignments: D18S16 (18p11.31), D18S12 (18p11.1), D18S1 (18q12.2), D18S13 (18q21.31), D18S18 (18q21.31), D18S14 (18q21.33), and D18S17 (18q23). In addition, the localization of D18S3 in 18p11.3 was confirmed and that of D18S6, previously mapped in 18p11, was changed to 18q21.13. Finally, a more accurate mapping for nine probes was proposed: D18S7 (18q12.2), D18S10 (18q12.2), D18S24 (18q21.13), D18S8 (18q21.13), GRP(18q21.31), BCL2(18q21.33), D18S5(18q22.1), D18S19 (18q22.1), and D18S11 (18q22.3). The ordering of probes located in close proximity was made possible by combined use of single-color FISH with direct assignment on banded chromosomes, chromosomal length measurements, and double-color FISH.
Chromosome assignment of human brain expressed sequence tags (ESTs) by analyzing fluorescently labeled PCR products from hybrid cell panels
Authors: Durkin A.S. , Nierman W.C. , Zoghbi H. , Jones C. , Kozak C.A. , Maglott D.R.
Published: May 2008

Sixty-three human brain cDNA sequences were newly assigned to individual human chromosomes. Ten of these were subregionally localized, and one was also mapped in the mouse genome. Four previously reported assignments were refined. PCR primers were designed from expressed sequence tags (ESTs) and tested for specific amplification from human genomic DNA. DNA was then amplified, often in multiplexed PCR reactions, using DNA from somatic cell hybrid mapping panels as templates. The amplification products were identified using an automated fluorescence detection system. Chromosomal assignments were made by discordancy analysis. Thirteen newly localized cDNAs exhibited homology to previously reported sequences. EST01471 was shown to correspond to human microtubule-associated protein 1B (MAP1B), confirming the previous mapping of this gene to human chromosome 5. Other genes tentatively assigned to chromosomes based on these results were a component of the signal peptide receptor of the endoplasmic reticulum (EST00745) and a cyclic AMP-regulated phosphoprotein (EST01041) on chromosome 1, a protein phosphatase 2A 55-kDa regulatory subunit (EST01650) on chromosome 4, an NAD(P) transhydrogenase (EST01744) on chromosome 5, ribosomal proteins LI a or Lib (EST01627) and L18a (EST01583), a brain transcription factor (BF-1, EST00795) on chromosome 14, a milk fat globule membrane-related protein (EST01678) on chromosome 15, a putative peptide initiation factor (EST00675) on chromosome 17, thiosulfate sulfurtransferase (TST) on chromosome 22, and moesin (MSN) (EST00896) and a human equivalent of rat spot 14 (S14) (EST00887) on Xp11→cen and Xpter→p21.3, respectively.