Spectral karyotyping (SKY) of mouse meiotic chromosomes

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 293-298 ◽  
Author(s):  
Henry HQ Heng ◽  
Guo Liu ◽  
Wei Lu ◽  
Steve Bremer ◽  
Christine J Ye ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Synaptonemal Complex ◽  
Meiotic Prophase ◽  
Spectral Karyotyping ◽  
Mitotic Chromosomes ◽  
Meiotic Chromosomes ◽  
Y Chromosomes ◽  
Behavioural Characteristics ◽  
Co Detection

The spectral karyotyping procedure of in situ hybridization with chromosome-specific probes assigns a unique colour code to each of the 21 mouse mitotic chromosomes. We have adapted this procedure to meiotic prophase chromosomes, and the results show that each of the pachytene or metaphase I bivalents can be identified. This technique has the potential to recognize synaptic anomalies and chromosome-specific structural and behavioural characteristics. We confirm these potentials by the recognition of the heterologous synapsis of the X and Y chromosomes and by the variances of synaptonemal complex lengths for each of the colour-coded bivalents in eight prophase nuclei.Key words: SKY, meiosis, synaptonemal complex, multicolour, chromosome painting, spectral karyotyping, protein-SKY co-detection.

Identical megabase transgenes on mouse chromosomes 3 and 4 do not promote ectopic pairing or synapsis at meiosis

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 770-773 ◽  
Author(s):  
P. B. Moens ◽  
J. A. M. Heddle ◽  
B. Spyropoulos ◽  
H. H. Q. Heng
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Synaptonemal Complex ◽  
Meiotic Prophase ◽  
Nuclear Volume ◽  
Dna Interactions ◽  
Synaptonemal Complexes ◽  
Meiotic Chromosomes ◽  
Ectopic Pairing

To investigate ectopic interactions at the chromatin level, we examined the meiotic organization of 1–2 mb phage λ transgenes on mouse chromosomes 3 and 4 by fluorescence in situ hybridization in combination with immunocytology of meiotic chromosomes. At early meiotic prophase, the transgenes are sufficiently dispersed in the nuclear volume to permit potential DNA–DNA interactions, but no synaptonemal complexes form between the sites of transgenes residing on different chromosomes. At later stages, when the chromatin is more condensed, the transgenes on different chromosomes are not preferentially associated as they are when they are on the same chromosome. At diplotene and metaphase I, no formations were observed that could be interpreted as reciprocal crossovers or chiasmata between the transgenes located on chromosomes 3 and 4. It appears that in normal fertile mice, a 1- to 2-mb homology is insufficient to initiate synapsis between nonhomologs, and it is concluded that homology is assessed within the broader context of the chromosome to initiate synapsis at meiotic prophase.Key words: transgenes, ectopic pairing, meiosis, synaptonemal complex, immunocytology, FISH.

scholarly journals Lateral Elements Inside Synaptonemal Complex-Like Polycomplexes in ndt80 Mutants of Yeast Bind DNA

Genetics ◽  
2003 ◽  
Vol 163 (2) ◽  
pp. 539-544 ◽  
Author(s):  
Hasanuzzaman Bhuiyan ◽  
Gunilla Dahlfors ◽  
Karin Schmekel
Keyword(s):  
In Situ Hybridization ◽  
Electron Microscope ◽  
Synaptonemal Complex ◽  
Immunoelectron Microscopy ◽  
Meiotic Prophase ◽  
Lateral Element ◽  
Homologous Chromosomes ◽  
Meiotic Prophase I ◽  
Dna Antibodies

Abstract The synaptonemal complex (SC) keeps the synapsed homologous chromosomes together during pachytene in meiotic prophase I. Structures that resemble stacks of SCs, polycomplexes, are sometimes found before or after pachytene. We have investigated ndt80 mutants of yeast, which arrest in pachytene. SCs appear normal in spread chromosome preparations, but are only occasionally found in intact nuclei examined in the electron microscope. Instead, large polycomplexes occur in almost every ndt80 mutant nucleus. Immunoelectron microscopy using DNA antibodies show strong preferential labeling to the lateral element parts of the polycomplexes. In situ hybridization using chromosome-specific probes confirms that the chromosomes in ndt80 mutants are paired and attached to the SCs. Our results suggest that polycomplexes can be involved in binding of chromosomes and possibly also in synapsis.

In situ localization of two repetitive DNA sequences to surface-spread pachytene chromosomes of rye

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 551-559 ◽  
Author(s):  
S. M. Albini ◽  
T. Schwarzacher
Keyword(s):  
In Situ Hybridization ◽  
Synaptonemal Complex ◽  
Dna Sequences ◽  
Meiotic Prophase ◽  
Hybridization Signal ◽  
Metaphase Chromosomes ◽  
Somatic Metaphase ◽  
Rdna Signal ◽  
Surface Spread

Surface-spread pollen mother cells at meiotic prophase from Secale cereale (rye) were used for fluorescent DNA:DNA in situ localization of two tandemly repeated DNA sequences: pTa71, a wheat rDNA clone, and pSc119.2, a cloned 120-bp repeat from rye heterochromatin. The fluorescent hybridization signal, consisting of many yellow-green dots, was closely associated with the bivalent axes, corresponding to the synaptonemal complex, and located in the surrounding chromatin. The rDNA signal was associated with one bivalent, the smallest of the seven, at a distance about 13% of the bivalent length from the telomere. This corresponded to the position of the nucleolar organizing region of silver-stained synaptonemal complexes analyzed under the electron microscope and published data for somatic metaphase chromosomes. The relative length of the axis covered with the rDNA signal is less than expected from somatic metaphases, but it corresponds more closely to the proportion of the sequences in the genome. The hybridization signal with the 120-bp repeat was located mainly at the telomeric regions of several bivalents that showed thickenings of the axis after DAPI staining, probably corresponding to somatic C-bands. These major and some minor intercalary sites agree with the distribution of the 120-bp repeat in somatic metaphase. Fluorescent in situ hybridization to plant surface-spread pachytene chromosomes, which can be obtained in large numbers, has great potential for studying meiotic prophase, high-resolution mapping of DNA sequences, and investigating the relationship of DNA sequences to the synaptonemal complex.Key words: in situ hybridization, cereals, pachytene, meiosis, synaptonemal complex, physical mapping.

scholarly journals Expression and localization of the mRNA for DNA (cytosine-5)- methyltransferase in mouse seminiferous tubules.

1994 ◽  
Vol 42 (9) ◽  
pp. 1271-1276 ◽  
Author(s):  
M Numata ◽  
T Ono ◽  
S Iseki
Keyword(s):  
In Situ Hybridization ◽  
Significant Role ◽  
Meiotic Prophase ◽  
Oligonucleotide Probe ◽  
Mouse Testis ◽  
Seminiferous Tubules ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry

DNA (cytosine-5)-methyltransferase (DNA MTase) is the only enzyme known to be involved in the methylation of mammalian DNA. Although the expression of DNA MTase gene is abundant in the testis, little is known about the role of this enzyme during spermatogenesis. We examined the distribution of DNA MTase mRNA in mouse testis by in situ hybridization histochemistry with an oligonucleotide probe. The mRNA signal was observed in the seminiferous tubules and was localized predominantly in spermatogonia and spermatocytes, particularly during the earlier steps of meiotic prophase I, with maximal intensity in the early pachytene cells. These results suggest some significant role for DNA MTase in spermatogenesis.

scholarly journals Application of the FISH Technique to Visualize Sex Chromosomes in Domestic Cat Spermatozoa

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2106
Author(s):  
Barbara Kij-Mitka ◽  
Halina Cernohorska ◽  
Svatava Kubickova ◽  
Sylwia Prochowska ◽  
Wojciech Niżański ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Sex Chromosomes ◽  
Chromosomal Abnormalities ◽  
Molecular Cytogenetics ◽  
Molecular Probes ◽  
Domestic Cat ◽  
Fish Technique ◽  
Y Chromosomes

Fluorescence in situ hybridization is a molecular cytogenetics technique that enables the visualization of chromosomes in cells via fluorescently labeled molecular probes specific to selected chromosomes. Despite difficulties in carrying out the FISH technique on sperm, related to the need for proper nuclear chromatin decondensation, this technique has already been used to visualize chromosomes in human, mouse, cattle, swine, horse, and dog spermatozoa. Until now, FISH has not been performed on domestic cat sperm; therefore, the aim of this study was to visualize sex chromosomes in domestic cat sperm. The results showed the presence of X and Y chromosomes in feline spermatozoa. The procedure used for sperm decondensation and fluorescence in situ hybridization was adequate to visualize chromosomes in domestic cat spermatozoa and, in the future, it may be used to determine the degree of chromosomal abnormalities in these gametes.

The testis-specific transcript (ferT) of the tyrosine kinase FER is expressed during spermatogenesis in a stage-specific manner

1990 ◽  
Vol 10 (9) ◽  
pp. 5021-5025
Author(s):  
E Keshet ◽  
A Itin ◽  
K Fischman ◽  
U Nir
Keyword(s):  
In Situ Hybridization ◽  
Tyrosine Kinase ◽  
Tyrosine Kinases ◽  
Meiotic Prophase ◽  
Hybridization Analysis ◽  
Pachytene Stage ◽  
Specific Transcript ◽  
Specific Manner

ferT is a testis-specific transcript of FER encoding a truncated version of the potential tyrosine kinase. Using in situ hybridization analysis, we found that ferT was transiently expressed during spermatogenesis and that expression was restricted to spermatocytes at the pachytene stage of meiotic prophase. This pattern of expression is unprecedented by other tyrosine kinases and suggests a role for ferT in a particular stage of spermatogenesis.

Identification and characterization of normal length nonfluorescent Y chromosomes: cytogenetic analysis, Southern hybridization and non-isotopic in situ hybridization

1990 ◽  
Vol 85 (6) ◽  
pp. 569-575 ◽  
Author(s):  
Frank Speleman ◽  
Bart Van der Auwera ◽  
Kathelijne Mangelschots ◽  
Miet Vercruyssen ◽  
Ton Raap ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Cytogenetic Analysis ◽  
Southern Hybridization ◽  
Normal Length ◽  
Y Chromosomes ◽  
Identification And Characterization

Genomic in situ hybridization analysis of a trigenomic hybrid involving Solanum and Lycopersicon species

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 299-304 ◽  
Author(s):  
S N Haider Ali ◽  
Dirk Jan Huigen ◽  
M S Ramanna ◽  
Evert Jacobsen ◽  
Richard GF Visser
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Doubling ◽  
Genomic In Situ Hybridization ◽  
Potato Genome ◽  
Meiotic Chromosomes ◽  
Lycopersicon Species ◽  
Bridge Species ◽  
Genomic Probes ◽  
Homoeologous Chromosomes

A 4x potato (+) tomato fusion hybrid (2n = 4x = 48) was successfully backcrossed with a diploid Lycopersicon pennellii (2n = 2x = 24). Genomic in situ hybridization (GISH) on somatic and meiotic chromosomes confirmed that the progenies were triploids (2n = 3x = 36) and possessed three different genomes: potato, tomato, and L. pennellii. Therefore, they have been called trigenomic hybrids. Total genomic probes of both Lycopersicon species were found to hybridize mutually, whereas the potato genome was clearly differentiated. During metaphase I, bivalents were formed predominantly between tomato and L. pennellii chromosomes and the univalents of potato chromosomes were most common. Trivalents in all cases included homoeologous chromosomes of potato, tomato, and L. pennellii. However, the triploids were totally sterile as determined from extensive crossing. On chromosome doubling of triploids by shoot regeneration from callus, hexaploids (2n = 6x = 72) were obtained. Despite exhibiting clear allohexaploid behaviour by forming 36 bivalents at meiosis, these were also completely sterile like their triploid counterparts. In spite of this drawback, the prospects of chromosome pairing between potato L. pennellii and Solanum genomes does open the possibilities for bringing the two genera close.Key words: trigenomic triploids, GISH, bridge species, potato (+) tomato fusion hybrids.

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