Localization of the lampbrush loop pair Nooses on the Y chromosome of Drosophila hydei by fluorescence in situ hybridization

Chromosoma ◽  
1993 ◽  
Vol 102 (8) ◽  
pp. 546-552 ◽  
Author(s):  
Ron Hochstenbach ◽  
Monique Wilbrink ◽  
Ron Suijkerbuijk ◽  
Wolfgang Hennig
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Drosophila Hydei

scholarly journals Fluorescence in situ hybridization in diluted and flow cytometrically sorted boar spermatozoa using specific DNA direct probes labelled by nick translation

Reproduction ◽  
2003 ◽  
pp. 317-325 ◽  
Author(s):  
I Parrilla ◽  
JM Vazquez ◽  
M Oliver-Bonet ◽  
J Navarro ◽  
J Yelamos ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Internal Control ◽  
Dna Probes ◽  
Chromosome 1 ◽  
Nick Translation ◽  
Fluorescent Signal ◽  
Sperm Separation

Successful evaluation of X- and Y-chromosome-bearing sperm separation technology using flow cytometry-cell sorter is of great importance. Fluorescence in situ hybridization (FISH), which allows for the detection of specific nucleic acid sequences on morphologically preserved spermatozoa, is an ideal method for quantitatively and qualitatively assessing the purity of sorted sperm samples. In this study specific pig DNA direct probes for small regions of chromosomes 1 and Y were used. Chromosome 1 was labelled in green and used as internal control to detect a lack of hybridization, whereas chromosome Y was labelled in red. Nick translation was used as the labelling method for the preparation of these probes. Spermatozoa, unsorted and sorted for high and low Y-chromosome purity from ejaculates of five boars, were fixed on slides and two-colour direct FISH was performed for chromosomes 1 and Y. About 500 non-sorted and 200 sorted spermatozoa per sample were scored. The proportion of Y-chromosome-bearing spermatozoa was determined by the presence of a red fluorescent signal on the sperm head and the proportion of X-chromosome-bearing spermatozoa was determined by subtraction. The efficiency of the hybridization procedure was established as near 98% on sorted and unsorted samples. The results of this study confirm that direct FISH using specific pig DNA probes labelled by nick translation provides a useful tool for laboratory validation of sperm separation by flow sorting technology. Moreover, the ease of nick translation and the quality of the fluorescent signal obtained using this method makes this procedure the most appropriate method for labelling pig DNA probes to be used for direct FISH on pig spermatozoa.

scholarly journals Prenatal Confirmation of the Translocation between Chromosome 15 and Y-Chromosome by Fluorescence in situ Hybridization.

1999 ◽  
Vol 187 (3) ◽  
pp. 285-289 ◽  
Author(s):  
Yukihito Fukada ◽  
Takehiko Yasumizu ◽  
Atsuhito Amemiya ◽  
Keiko Kohno ◽  
Motoi Takizawa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Chromosome 15

Fluorescence in situ hybridization with y chromosome-specific probe in decondensed bovine spermatozoa

1999 ◽  
Vol 52 (6) ◽  
pp. 1043-1054 ◽  
Author(s):  
J. Kobayashi ◽  
T. Kohsaka ◽  
H. Sasada ◽  
M. Umezu ◽  
E. Sato
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Specific Probe ◽  
Bovine Spermatozoa

RAPD markers encoding retrotransposable elements are linked to the male sex in Cannabis sativa L.

Genome ◽  
2005 ◽  
Vol 48 (5) ◽  
pp. 931-936 ◽  
Author(s):  
Koichi Sakamoto ◽  
Tomoko Abe ◽  
Tomoki Matsuyama ◽  
Shigeo Yoshida ◽  
Nobuko Ohmido ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Dna Sequences ◽  
Blot Analysis ◽  
Cannabis Sativa ◽  
Random Amplified Polymorphic Dna ◽  
Male And Female ◽  
Retrotransposable Elements

Male-associated DNA sequences were analyzed in Cannabis sativa L. (hemp), a dioecious plant with heteromorphic sex chromosomes. DNA was isolated from male and female plants and subjected to random amplified polymorphic DNA analysis. Of 120 primers, 17 yielded 400 to 1500-bp fragments detectable in male, but not female, plants. These fragments were cloned and used as probes in gel-blot analysis of genomic DNA. When male and female DNA was hybridized with 2 of these male-specific fragments, MADC(male-associated DNA sequences in C. sativa)3 and MADC4, particularly intense bands specific to male plants were detected in addition to bands common to both sexes. The MADC3 and MADC4 sequences were shown to encode gag/pol polyproteins of copia-like retrotransposons. Fluorescence in situ hybridization with MADC3 and MADC4 as probes revealed a number of intense signals on the Y chromosome as well as dispersed signals on all chromosomes. The gel-blot analysis and fluorescence in situ hybridization results presented here support the hypothesis that accumulation of retrotransposable elements on the Y chromosome might be 1 cause of heteromorphism of sex chromosomes.Key words: Cannabis sativa, FISH, RAPD, retrotransposon, sex chromosome.

scholarly journals An X-Y paint set and sperm FISH protocol that can be used for validation of cattle sperm separation procedures

Reproduction ◽  
2001 ◽  
pp. 541-546 ◽  
Author(s):  
W Rens ◽  
F Yang ◽  
G Welch ◽  
S Revell ◽  
PC O'Brien ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Close Agreement ◽  
Fish Method ◽  
Sperm Fish ◽  
Sperm Separation ◽  
Sperm Decondensation

X and Y chromosome paints were developed from sorted yak chromosomes for sexing cattle spermatozoa. Clear hybridization signals were obtained for every spermatozoon using a modified sperm decondensation protocol and fluorescence in situ hybridization (FISH). The procedure was evaluated using the established Beltsville sperm sexing technology, which separates spermatozoa by flow cytometry into X- and Y-bearing fractions. Close agreement was found between the assessment of sperm separation by flow cytometry and by FISH with the X-Y paint set. The FISH method is a simple, reliable and robust procedure for assessing the effectiveness of separation of X and Y spermatozoa.

scholarly journals Fluorescence In Situ Hybridization for the X and Y Chromosome Centromeres Helps Differentiate Between Gestational and Nongestational Choriocarcinoma in Clinically Ambiguous Cases

2019 ◽  
Vol 144 (7) ◽  
pp. 863-868
Author(s):  
Rumeal D. Whaley ◽  
Rachel E. Dougherty ◽  
Liang Cheng ◽  
Thomas M. Ulbright
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
Genetic Material ◽  
Gestational Trophoblastic Disease ◽  
Definitive Diagnosis ◽  
Molecular Genotyping ◽  
Trophoblastic Disease ◽  
Gestational Choriocarcinoma

Context.— Gestational choriocarcinoma usually presents during the reproductive years, typically within 1 year of pregnancy, although presentation remote from pregnancy also occurs and may cause confusion with other tumors, including choriocarcinoma of germ cell origin and somatic carcinomas with choriocarcinomatous differentiation. It is important to separate these tumors for treatment and prognostic reasons. Objective.— To assess the utility of fluorescence in situ hybridization for the X and Y chromosome centromeres in determining the gestational origin of clinically ambiguous extrauterine choriocarcinomas in women. Design.— A review of female patients with extrauterine choriocarcinomas who had no evidence of prior gestational trophoblastic disease was performed. Samples were analyzed by fluorescence in situ hybridization for the X and Y chromosome centromeres using standard methodologies. Results.— Five cases met the criteria, all of which displayed trophoblastic cells and necrosis. Three cases (60%) had Y chromosomes by fluorescence in situ hybridization, which confirmed gestational origin. Although the 2 cases without a Y chromosome would ordinarily require molecular genotyping for paternal genetic material to establish gestational origin, in one of these cases a subsequent recurrence of yolk sac tumor allowed confirmation of its mediastinal origin. Conclusions.— Fluorescence in situ hybridization for detection of the X and Y chromosome centromeres is an effective screening test for gestational choriocarcinoma. It provided a definitive diagnosis of metastatic gestational choriocarcinoma in 3 of 5 potential cases that lacked a clinical history of gestational trophoblastic disease. An additional benefit is that more laboratories have the capability to perform fluorescence in situ hybridization than can perform molecular genotyping for definitive diagnosis.

A multicenter investigation with interphase fluorescence in situ hybridization using X- and Y-chromosome probes

1998 ◽  
Vol 76 (4) ◽  
pp. 318-326 ◽  
Author(s):  
Gordon Dewald ◽  
Richard Stallard ◽  
A. Al Saadi ◽  
Susan Arnold ◽  
Patricia I. Bader ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome

268 IDENTIFICATION OF X- AND Y-BEARING SPERMATOZOA IN SORTED BUFFALO (BUBALUS BUBALIS) SEMEN BY FLUORESCENCE IN SITU HYBRIDIZATION

2009 ◽  
Vol 21 (1) ◽  
pp. 231
Author(s):  
M. Zhang ◽  
X. J. Zhuang ◽  
Y. Q. Lu ◽  
C. H. Hu ◽  
S. S. Lu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Y Chromosome ◽  
X Chromosome ◽  
Chromosome Painting ◽  
Fish Method ◽  
Painting Probes ◽  
Flow Cytometry Sorting

Flow cytometry sorting technology has been successfully used to sort the X- and Y-chromosome bearing sperm. Previous studies showed that fluorescence in situ hybridization (FISH) method was a simple and reliable procedure for assessing the effectiveness of separation of X- and Y-sperm in the swine (Kawarasaki T et al. 1998 Theriogenology 50, 625–635) and the bovine (Rens W et al. 2001 Reproduction 121, 541–546). Reports of sex-preselection by flow-cytometry sorting of the X- and Y-sperm were also seen in the buffalo (Presicce GA et al. 2005 Reprod. Dom. Anim. 40, 73–75; Lu YQ et al. 2006 Anim. Reprod. Sci. 100, 192–196). There was, however, no report to date for using the FISH method to assess the purity of the sorted buffalo sperm. The objective of the present study was to verify the purity of flow cytometrically-sorted buffalo X- and Y-sperm by FISH using bovine X- and Y- chromosome painting probes prepared by microdissection. The X- and Y- chromosomes of bovidea were microdissected respectively from the metaphase spreads of Holstein blood cells with a glass needle controlled by a micromanipulator and amplified by degenerate oligo-nucleotide primer-PCR (DOP-PCR) (Mariela N et al. 2005 Genet. Mol. Res. 4, 675–683). The DOP-PCR products of X- and Y- chromosome were labeled with CY3-dUTP and Biotin-11-dUTP, respectively. The buffalo X- or Y-sperm DNA from unsorted semen and sorted semen were hybridized to the labeled probes, respectively. The results showed that the hybridized signals were clearly visible in the metaphase karyotype of bovine and buffalo semen samples. About 47.7% (594/1246) and 48.9% (683/1396) of the unsorted buffalo sperm emitted strong fluorescent signals when assessed by Y- and X-chromosome painting probes, respectively, which was conformed to the sex ratio in normal buffalo sperm (50%:50%). About 86.1% (1529/1776) hybridization signals of the sperm in the sorted X-semen assessed by X-chromosome painting probes were detected, while 82.2% (2232/2716) of the Y-sorted buffalo sperm emitted strong fluorescent signals when assessed by Y-chromosome painting probe. The results of the flow cytometer re-analysis revealed that the proportions of X- and Y-bearing sperm in the sorted semen were 89.6% and 86.7%, respectively. There were no apparent differences between the two assessment methods of sperm separation by flow cytometry re-analysis and by FISH with the X-Y paint probe. In conclusion, bovine X- and Y-chromosome painting probes prepared using microisolation method could be used to verify the purity of the sorted sperm in the buffalo. This study was supported by the Guangxi Department of Science and Technology (0626001-3-1) and National Key Technology R&D Program, The People’s Republic of China (2006BAD04A18). The authors (M. Zhang, X.J. Zhuang, and Y.Q. Lu) contributed equally to this work.

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