scholarly journals Multicolor FISH Analysis of Chromosomal Breaks, Duplications, Deletions, and Numerical Abnormalities in the Sperm of Healthy Men

2000 ◽  
Vol 67 (4) ◽  
pp. 862-872 ◽  
Author(s):  
Eddie D. Sloter ◽  
Xiu Lowe ◽  
Dan H. Moore ◽  
Joginder Nath ◽  
Andrew J. Wyrobek
Keyword(s):  
Fish Analysis ◽  
Multicolor Fish ◽  
Healthy Men ◽  
Chromosomal Breaks

Integrated Karyotyping of Woodland Strawberry (Fragaria vesca) with Oligopaint FISH Probes

2017 ◽  
Vol 153 (3) ◽  
pp. 158-164 ◽  
Author(s):  
Manman Qu ◽  
Kunpeng Li ◽  
Yanli Han ◽  
Lei Chen ◽  
Zongyun Li ◽  
...  
Keyword(s):  
Draft Genome ◽  
5S Rdna ◽  
Fish Analysis ◽  
Fragaria Vesca ◽  
Chromosome Identification ◽  
Multicolor Fish ◽  
Metaphase Chromosomes ◽  
Cross Species Chromosome Painting ◽  
Simultaneous Identification

Chromosome identification is critical for many aspects of cytogenetic research. However, for Fragaria vesca, definite identification of individual chromosomes is almost impossible because of their small size and high similarity. Here, we demonstrate that bulked oligonucleotide (oligo) probes can be used as chromosome-specific DNA markers for chromosome identification in F. vesca. Oligos specific to entire pseudochromosomes in the draft genome of F. vesca were identified and synthesized as libraries. In all, we synthesized 6 oligo libraries corresponding to 6 pseudochromosomes of F. vesca. These libraries were amplified and labeled as probes for fluorescence in situ hybridization (FISH). Two rounds of multicolor FISH analysis were sequentially conducted on the same metaphase cells with each round including 3 probe libraries, which permitted simultaneous identification of all chromosomes of F. vesca. Moreover, 45S and 5S rDNA were mapped to chromosomes 1, 2, and 7, respectively. A karyotype of metaphase chromosomes was constructed, representing the first FISH-based molecular cytogenetic karyotype of F. vesca. Our study can serve as a basis for future comparative cytogenetic research through cross-species chromosome painting using bulked oligo probes and will facilitate the application of breeding technologies that rely on the identification of chromosomes in the genus Fragaria.

scholarly journals Molecular Cytogenetic Mapping of Chromosomal Fragments and Immunostaining of Kinetochore Proteins in Beta

2009 ◽  
Vol 2009 ◽  
pp. 1-12 ◽  
Author(s):  
Daryna Dechyeva ◽  
Thomas Schmidt
Keyword(s):  
Repetitive Dna ◽  
Small Chromosome ◽  
Rrna Genes ◽  
Fish Analysis ◽  
Addition Lines ◽  
Multicolor Fish ◽  
Active Involvement ◽  
Chromosome Fragments ◽  
In The Wild ◽  
Wild Beet

By comparative multicolor FISH, we have physically mapped small chromosome fragments in the sugar beet addition lines PRO1 and PAT2 and analyzed the distribution of repetitive DNA families in species of the section Procumbentes of the genus Beta. Six repetitive probes were applied, including genotype-specific probes—satellites pTS4.1, pTS5, and pRp34 and a dispersed repeat pAp4, the telomere (TTTAGGG)n, and the conserved 18S-5.8S-25S rRNA genes. Pachytene-FISH analysis of the native centromere organization allowed proposing the origin of PRO1 and PAT2 fragments. Comparative analysis of the repetitive DNA distribution and organization in the wild beet and in the addition lines allowed the development of a physical model of the chromosomal fragments. Immunostaining revealed that the PRO1 chromosome fragment binds α-tubulin and the serine 10-phosphorylated histone H3 specific for the active centromere. This is the first experimental detection of the kinetochore proteins in Beta showing their active involvement in chromosome segregation in mitosis.

scholarly journals Coexpression of Three Odorant-Binding Protein Genes in the Foreleg Gustatory Sensilla of Swallowtail Butterfly Visualized by Multicolor FISH Analysis

2021 ◽  
Vol 1 ◽  
Author(s):  
Atsushi Ugajin ◽  
Katsuhisa Ozaki
Keyword(s):  
Host Plants ◽  
Fish Analysis ◽  
Multicolor Fish ◽  
Plant Recognition ◽  
Lepidopteran Insects ◽  
Soluble Ligand ◽  
Multicolor Fluorescence ◽  
Odorant Binding ◽  
Gustatory Sensilla

Lepidopteran insects are mostly monophagous or oligophagous. Female butterflies distinguish their host plants by detecting a combination of specific phytochemicals through the gustatory sensilla densely distributed on their foreleg tarsi, thereby ensuring oviposition on appropriate host plants. In this study, to gain insight into the molecular mechanism underlying host plant recognition by the gustatory sensilla, using Asian swallowtail, Papilio xuthus, we focused on a family of small soluble ligand-binding molecules, odorant-binding proteins (OBPs), and found that three OBP genes showed enriched expression in the foreleg tarsus. Multicolor fluorescence in situ hybridization analyses demonstrated the coexpression of these three OBP genes at the bases of the foreleg gustatory sensilla. Further analyses on other appendages revealed that PxutOBP3 was exclusively expressed in the tissues which could have direct contact with the leaf surface, suggesting that this OBP gene specifically plays an important role in phytochemicals perception.

Multicolor FISH analysis of rDNA and telomere on spin-ach

2007 ◽  
Vol 29 (11) ◽  
pp. 1405
Author(s):  
Tian-Ying LAN
Keyword(s):  
Fish Analysis ◽  
Multicolor Fish

Multicolor-FISH analysis of a natural killer cell line (NK-92)

2002 ◽  
Vol 26 (11) ◽  
pp. 1027-1033 ◽  
Author(s):  
Roderick A.F MacLeod ◽  
Stefan Nagel ◽  
Maren Kaufmann ◽  
Karin Greulich-Bode ◽  
Hans G Drexler
Keyword(s):  
Natural Killer Cell ◽  
Natural Killer ◽  
Cell Line ◽  
Killer Cell ◽  
Fish Analysis ◽  
Multicolor Fish ◽  
Natural Killer Cell Line

Multicolor FISH analysis of rDNA and telomere on spinach

2008 ◽  
Vol 2 (3) ◽  
pp. 314-316
Author(s):  
Tianying Lan ◽  
Bo Liu ◽  
Fengping Dong ◽  
Ruiyang Chen ◽  
Xiulan Li ◽  
...  
Keyword(s):  
Fish Analysis ◽  
Multicolor Fish

Proof of Y-Loss As Clonal Abnormality in MDS by Comparative Analysis of CD34+ and CD3+ Peripheral Blood Cells,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3800-3800 ◽  
Author(s):  
Christina Ganster ◽  
Friederike Braulke ◽  
Katayoon Shirneshan ◽  
Dietrich Kämpfe ◽  
Uwe Platzbecker ◽  
...  
Keyword(s):  
T Cells ◽  
Y Chromosome ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Fish Analysis ◽  
Peripheral Blood Cells ◽  
Clone Size ◽  
Healthy Men ◽  
Age Related ◽  
Cd34 Cells

Abstract Abstract 3800 Introduction: In an ongoing diagnostic study we are currently following chromosomal anomalies in immunomagnetically enriched CD34+ peripheral blood cells in patients with suspected or proven myelodysplastic syndromes (MDS) at short intervals using fluorescence in situ hybridization (FISH) analysis every two to three months over three years. A loss of the Y chromosome was detected in 4% of these patients, as a single anomaly in 2%. Since it is controversially discussed whether loss of the Y chromosome is an age-related or a clonal event in patients with MDS, we aimed to examine whether a Y-loss is clonal or an age-related event in our patients. Methods: For patients with known Y-loss, we used peripheral blood not only to immunomagnetically enrich clonal CD34+ cells, but also CD3+ T-cells not belonging to the MDS clone. Subsequently, we performed FISH analysis to compare the clone sizes of cells with Y-loss in CD34+ and CD3+ cells. As our laboratory threshold for the FISH probe in CD34+ peripheral blood cells is 5%, we included 18 patients with clone sizes exceeding this threshold in CD34+ cells. The median age of the patients was 76 years (range 62–89). To establish a laboratory threshold for the FISH probe in CD3+ peripheral blood cells, we analyzed T-cells of 25 healthy men with a median age of 27 years (range 19–35). Furthermore, we just initialized an investigation of the laboratory threshold for the FISH probe in CD3+ peripheral blood cells of elder men by measuring the frequency of loss of the Y chromosome in T-cells of this control cohort not suffering from hematopoietic diseases. Until now we could recruit 15 men with a median age of 75 years (range 66–84) for this purpose, further will follow soon. Results: In patients with suspected or proven MDS, the number of cells with -Y was significantly increased in CD34+ cells compared to CD3+ cells (p<0.0001). The median clone size was 64% (range 12–97) in CD34+ cells and 5% (range 1–14) in CD3+ T-cells. The clone size in CD34+ cells was at least four times higher than in CD3+ cells in all patients. We could not detect further chromosomal abnormalities in 16 patients. Chromosomal banding analysis revealed that cells with -Y and cells with -Y and +8 occurred in parallel in two patients. In men below the age of 35 Y-loss could not be detected. The median clone size of 0.5% (range 0–2) resulted in a laboratory threshold of 2%. Interim analysis of men over the age of 65 resulted in a median clone size of 2.5% (range 1–14) and a laboratory threshold of 13%. So far the FISH-signal corresponding to the Y chromosome was significantly more frequent missing in T-cells of elder than in T-cells of younger men (p=0.005). Conclusion: Regarding the absence of Y-loss in CD3+ peripheral blood cells of young healthy men compared to up to 14% -Y in CD3+ peripheral blood cells of elder men, the low proportion of -Y in CD3+ cells of our patients suggests an age related Y-loss in normal T-cells. As the number of CD34+ peripheral blood cells with -Y exceeds the number of CD3+ peripheral blood cells with -Y in all patients, we assume that Y-loss is clonal to some extent in all of them. We established a reliable method to test if loss of the Y chromosome is disease- or age-related in individual MDS patients. It can be used to determine a clonal disease in patients with suspected MDS and Y-loss as sole abnormality. Disclosures: No relevant conflicts of interest to declare.

CFTR protein expression in healthy men spermatozoa is not correlated with fertilization rate of ova

2010 ◽  
Vol 34 (8) ◽  
pp. S12-S12
Author(s):  
Hong‑Ge Li ◽  
Chen Min Xu ◽  
Kun Li ◽  
Ya Ni ◽  
Wen‑Ying Chen ◽  
...  
Keyword(s):  
Protein Expression ◽  
Fertilization Rate ◽  
Healthy Men ◽  
Cftr Protein
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