scholarly journals Application of Veterinary Cytogenetics in Domestic Animals: A Review

2019 ◽  
pp. 1-16
Author(s):  
Muhammad Sanusi Yahaya ◽  
Mohd Shahrom Salisi ◽  
Nur Mahiza Md. Isa ◽  
Abd Wahid Haron ◽  
Innocent Damudu Peter
Keyword(s):  
Chromosomal Abnormalities ◽  
Hybrid Sterility ◽  
Molecular Cytogenetics ◽  
Basic Research ◽  
Animal Production ◽  
Somatic Development ◽  
Molecular Cytogenetic ◽  
Molecular Cytogenetic Techniques

Cytogenetics is the study of chromosomes; their structure and properties, chromosome behavior during cell division, their influence on traits and factors which cause changes in chromosomes.  Veterinary cytogenetics is the application of cytogenetics to clinical problems that occur in animal production. It has been applied to understand problems such as infertility and its types, embryonic and fetal death, abnormality in sexual and somatic development and hybrid sterility and also prenatal sex determination and other forms of chromosomal abnormalities. These are achieved through conventional and banded karyotyping techniques and molecular cytogenetic techniques. Although conventional techniques are still useful and very widely applied, the nature of cytogenetics has gradually changed as a result of advances achieved in the molecular cytogenetic techniques for example fluorescent in situ hybridization and array-based techniques. These changes are evident in both molecular diagnostics and basic research. The combination of conventional and molecular cytogenetics has given rise to high resolution techniques which have enabled the study of fundamental questions regarding biological processes. It enables the study of inherited syndromes, the mechanisms of tumorigenesis at molecular level, genome organization and the determination of chromosome homologies between species. It allows the ease with which animals are selected in breeding programs and other important aspects of animal production. In this paper we discussed a number of techniques employed in cytogenetics and their methodologies, and recommend where future focus should be for the benefits of animal production.

scholarly journals Establishment and Optimization of Molecular Cytogenetic Techniques (45S Rdna-FISH, GISH And Fiber-FISH) In Kiwifruit (Actinidia L.)

2021 ◽  
Author(s):  
Yang Zhao ◽  
Honghong Deng ◽  
Yao Chen ◽  
Jihan Li ◽  
Silei Chen ◽  
...  
Keyword(s):  
Fiber Quality ◽  
Molecular Cytogenetics ◽  
45S Rdna ◽  
Metaphase Chromosomes ◽  
Physical Size ◽  
Molecular Cytogenetic ◽  
Rdna Fish ◽  
Molecular Cytogenetic Techniques ◽  
First Time

Abstract Background: Kiwifruit has long been regarded as ‘the king of fruits’ for its nutritional importance. However, the molecular cytogenetics of kiwifruit has long been hampered because of the large number of basic chromosome (x=29), the inherent small size and highly similar morphology of metaphase chromosomes. Fluorescence in situ hybridization (FISH) is an indispensable molecular cytogenetic technique widely used in many plant species. Herein, the effects of post-hybridization washing temperature on FISH, blocking DNA concentration on genomic in situ hybridization (GISH), extraction method on nuclei isolation and the incubation time on the DNA fiber quality in kiwifruit were evaluated.Results: The post-hybridization washing in 2×SSC solution for 3×5 min at 37 ˚C ensured high stringency and distinct specific FISH signals in kiwifruit somatic chromosomes. The use of 50× blocking DNA provided an efficient and reliable means of discriminating between chromosomes derived from in the hybrids of A. chinensis var. chinensis (2n=2x=58) × A. eriantha Benth (2n=2x=58), and inferring the participation of parental genitors. The chopping method established in the present study were found to be very suitable for preparation of leaf nuclei in kiwifruit. A high-quality linear DNA fiber was achieved by an incubation of 20 min. The physical size of 45S rDNA signals was approximately 35-40 μmm revealed by the highly reproducible fiber-FISH procedures established and optimized in this study.Conclusions: The molecular cytogenetic techniques (45S rDNA-FISH, GISH, and high-resolution fiber-FISH) for kiwifruit was for the first time established and optimized in the present study, which is the foundation for the future genomic and evolutionary studies.

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.

scholarly journals Molecular cytogenetic studies in rubber, Hevea brasiliensis Muell. Arg. (Euphorbiaceae)

Genome ◽  
1998 ◽  
Vol 41 (3) ◽  
pp. 464-467 ◽  
Author(s):  
Andrew R Leitch ◽  
K Yoong Lim ◽  
Ilia J Leitch ◽  
Michelle O'Neill ◽  
MeeLen Chye ◽  
...  
Keyword(s):  
Genetic Mapping ◽  
Ribosomal Dna ◽  
Hevea Brasiliensis ◽  
Consensus Sequence ◽  
Molecular Cytogenetics ◽  
5S Rdna ◽  
Single Pair ◽  
Interphase Nuclei ◽  
Molecular Cytogenetic

This paper reports the start of a molecular cytogenetics programme targeting the genome of the angiosperm tree species Hevea brasiliensis Muell. Arg. (rubber, 2n = 36), a major world crop about whose genetics very little is known. A metaphase karyotype of rubber is presented. In situ hybidization with the probe pTa71 for ribosomal DNA (rDNA) shows that there are four sites of probe hybidization occurring on two pairs of chromosomes called chromosomes 6 and 7 carrying sites NOR-1 and NOR-2, respectively. An examination of meristematic interphase nuclei shows that all four loci have the potential to be partially decondensed at interphase, although in many nuclei one or more loci appear fully condensed and apparently inactive. The probe pXVI revealed a single pair of chromosomes carrying 5S rDNA. The probes pTa71 and pXVI provide cytogenetic markers for three pairs of chromosomes that will be of use in genetic mapping programmes. The rubber chromosomes also have telomere sequences that hybridize with the Arabidopsis consensus sequence TTTAGGG. With the exception of the satellite region containing rDNA, which fluoresces brightly with chromomycin A3, fluorescence banding showed that there is no strong demarcation of the genome into GC- and AT-rich regions, as occurs in mammalian genomes.Key words: rubber, Hevea, genetic mapping, cytogenetics, ribosomal DNA, rDNA fluorescence banding.

scholarly journals Contributions of Cytogenetics and Molecular Cytogenetics to the Diagnosis of Adipocytic Tumors

2011 ◽  
Vol 2011 ◽  
pp. 1-9 ◽  
Author(s):  
Jun Nishio
Keyword(s):  
Fusion Gene ◽  
Molecular Cytogenetics ◽  
Chromosomal Translocations ◽  
Molecular Cytogenetic ◽  
Mesenchymal Neoplasms ◽  
Adipocytic Tumors ◽  
Molecular Cytogenetic Techniques ◽  
Conventional Cytogenetics ◽  
Well Differentiated

Over the last 20 years, a number of tumor-specific chromosomal translocations and associated fusion genes have been identified for mesenchymal neoplasms including adipocytic tumors. The addition of molecular cytogenetic techniques, especially fluorescence in situ hybridization (FISH), has further enhanced the sensitivity and accuracy of detecting nonrandom chromosomal translocations and/or other rearrangements in adipocytic tumors. Indeed, most resent molecular cytogenetic analysis has demonstrated a translocation t(11;16)(q13;p13) that produces aC11orf95-MKL2fusion gene in chondroid lipoma. Additionally, it is well recognized that supernumerary ring and/or giant rod chromosomes are characteristic for atypical lipomatous tumor/well-differentiated liposarcoma and dedifferentiated liposarcoma, and amplification of 12q13–15 involving theMDM2,CDK4, andCPMgenes is shown by FISH in these tumors. Moreover, myxoid/round cell liposarcoma is characterized by a translocation t(12;16)(q13;p11) that fuses theDDIT3andFUSgenes. This paper provides an overview of the role of conventional cytogenetics and molecular cytogenetics in the diagnosis of adipocytic tumors.

Use of meiotic FISH for identification of a new monosome in Gossypium hirsutum L.

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 34-40 ◽  
Author(s):  
Yuanfu Ji ◽  
Dwaine A. Raska ◽  
M. Nurul Islam-Faridi ◽  
Charles F. Crane ◽  
Michael S. Zwick ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Gossypium Hirsutum ◽  
Fluorescence In Situ Hybridization ◽  
Molecular Cytogenetics ◽  
Clinical Diagnostics ◽  
Identification System ◽  
Gossypium Hirsutum L ◽  
Molecular Cytogenetic ◽  
Cytogenetic Methods

The extensive use of molecular cytogenetics in human genetics and clinical diagnostics indicates that analogous applications in plants are highly feasible. One sort of application would be the identification of new aneuploids, which traditionally involves either direct karyotypic identification, which is feasible in only a few plant species, or tests with markers (cytogenetic, genetic, or molecular), which require sexual hybridization and at least one subsequent seed or plant generation. We have used meiotic fluorescence in situ hybridization (FISH) to analyze a new monosome of cotton (Gossypium hirsutum L., 2n = 4x = 52, 2(AD)1) that had a phenotype which seemed to be distinct from monosomes in the Cotton Cytogenetic Collection. Painting with A2-genome DNA revealed the monosome's D-subgenome origin. DAPI–PI staining showed that the monosome carries a major NOR, delimiting it to the major NOR-bearing chromosomes of the D-subgenome, i.e., 16 or 23. Dual-color FISH with 5S and 18S–28S rDNAs indicated that the monosome contains separate major clusters of each of these two tandemly repeated rDNA elements, thus delimiting the monosome to chromosome 23, for which the Cotton Cytogenetic Collection has previously been devoid of any sort of deficiency. Of the 26 chromosomes in the cotton genome, the Collection now provides coverage for 16 (70%) in the form of monosomy, and 20 (77%) in the form of monosomy and (or) telosomy. Use of molecular cytogenetic methods to identify a new plant aneuploid in cotton exemplifies the fact that a physicochemical karyotypic chromosome identification system is not required a priori for application of new molecular cytogenetic methods, thus indicating their potential applicability to nearly all plant species.Key words: fluorescence in situ hybridization, monosome, aneuploid, Gossypium hirsutum.

scholarly journals Genotype-Phenotype Characterization of Wolf-Hirschhorn Syndrome Confirmed by FISH: Case Reports

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
F. Sheth ◽  
O. R. Akinde ◽  
C. Datar ◽  
O. V. Adeteye ◽  
J. Sheth
Keyword(s):  
Cytogenetic Study ◽  
Case Reports ◽  
Normal Karyotype ◽  
Comparative Genomic ◽  
Molecular Cytogenetic ◽  
Contiguous Gene ◽  
Multiple Malformation ◽  
Molecular Cytogenetic Techniques

The Wolf-Hirschhorn syndrome (WHS) is a multiple malformation and contiguous gene syndrome resulting from the deletion encompassing a 4p16.3 region. A microscopically visible terminal deletion on chromosome 4p (4p16→pter) was detected in Case 1 with full blown features of WHS. The second case which had an interstitial microdeletion encompassingWHSC 1andWHSC 2genes at 4p16.3 presented with less striking clinical features of WHS and had an apparently “normal” karyotype. The severity of the clinical presentation was as a result of haploinsufficiency and interaction with surrounding genes as well as mutations in modifier genes located outside the WHSCR regions. The study emphasized that an individual with a strong clinical suspicion of chromosomal abnormality and a normal conventional cytogenetic study should be further investigated using molecular cytogenetic techniques such as fluorescencein situhybridization (FISH) or array-comparative genomic hybridization (a-CGH).

scholarly journals Estudo da Prevalência de Anomalias Cromossómicas em Abortamentos Espontâneos ou Mortes Fetais

2014 ◽  
Vol 27 (1) ◽  
pp. 42 ◽  
Author(s):  
Raquel Bastos ◽  
Carla Ramalho ◽  
Sofia Dória
Keyword(s):  
Gestational Age ◽  
Maternal Age ◽  
Chromosomal Abnormalities ◽  
Cytogenetic Study ◽  
Accurate Determination ◽  
Normal Karyotype ◽  
First Trimester ◽  
Conventional Technique ◽  
Molecular Cytogenetic Techniques

<strong>Introduction:</strong> Spontaneous abortion is one of the most frequent problems of pregnancy, estimated to affect, at least, one in every four women who tries to conceive. The main purpose of this work was to study the prevalence of chromosomal abnormalities in gestational losses, evaluating their relation with maternal age, gestational age and previous abortion history.<br /><strong>Material and Methods:</strong> Retrospective study of 401 pregnancy loss cases that have gone through cytogenetic and anatomopathologic analysis between January 2008 and June 2012, in Centro Hospitalar de S. João, Oporto.<br /><strong>Results:</strong> Of the 401 cases sent to cytogenetic study, it was possible to obtain information about the chromosomal complement in 333 cases, of which 72.7% showed normal karyotype, and 27.3% abnormal karyotype. Aneuploidies represented 92.3% of the identified chromosomopathies, with trisomies being the most frequent, related with an advanced maternal age and an early gestational age. There was no agreement between the results of the cytogenetic and the anatomopathologic analysis.<br /><strong>Discussion/Conclusion: </strong>The prevalence of chromosomal abnormalities, during the first trimester, is similar between sporadic and recurrent miscarriages. With increased maternal age, trisomies, the most frequent type of aneuploidy, are more likely to occur, with a mean increment in probability of 7.4% per year. A significant karyotype-pathological correlation was not established. Maternal contamination is the main obstacle to the accurate determination of the prevalence of chromosomal abnormalities. The molecular cytogenetic techniques already available can overcome the limitations of the conventional technique.

scholarly journals The Search for Molecular Defects in Genetic Disease

1996 ◽  
Vol 45 (1-2) ◽  
pp. 43-52
Author(s):  
H. Galjaard
Keyword(s):  
Cell Biology ◽  
Chromosomal Abnormalities ◽  
Single Gene ◽  
Molecular Cytogenetics ◽  
Basic Research ◽  
Laboratory Diagnosis ◽  
Mendelian Inheritance ◽  
Congenital Disorders ◽  
Perinatal Complications ◽  
Increased Risk

In most wealthy industrialized countries, socioeconomic circumstances, hygiene and medical care have improved to such an extent that perinatal complications and congenital disorders have become the major causes of infant mortality and of chronic handicap in children [1].In the past four decades, we have made impressive advances understandig the cytogenetic and molecular basis of congenital disorders. Dozens of syndromes associated with multiple physical handicaps and mental retardation have been related to specific numerical or structural chromosome aberrations. In situ hybridization and the development of a rapidly increasing number of DNA probes enable the detection of chromosomal abnormalities with a resolution down to the level of a single gene. The discovery of tumor-associated chromosomal aberrations, microdeletions, more than a hundred oncogenes, and the cloning of tumor suppressor genes are illustrations of the importance of molecular cytogenetics.When I started my scientific career at the end of the fifties, some 1,000 syndromes were proven or suspected to be subject to Mendelian inheritance and this number has now increased to more than 6,600 [2]. Progress in elucidating the protein defects responsible for these single gene disorders has been much slower and currently about 400 protein defects have been delineated [3]. This knowledge has been important not only for basic research on biochemical pathways and cell biology, but has also provided new perspectives for early laboratory diagnosis of index patients, carrier detection, genetic counseling and in some instances, such as phenylketonuria, for newborn screening and early treatment [4]. In those instances where a protein defect is also expressed in cultured skin fibroblasts, amniotic cells or chorionic villi, prenatal diagnosis becomes possible for couples at increased risk [5, 6].

scholarly journals Molecular cytogenetic analysis in the study of brain tumors: findings and applications

2005 ◽  
Vol 19 (5) ◽  
pp. 1-36 ◽  
Author(s):  
Jane Bayani ◽  
Ajay Pandita ◽  
Jeremy A. Squire
Keyword(s):  
Brain Tumors ◽  
Cytogenetic Analysis ◽  
Molecular Cytogenetics ◽  
Response To Treatment ◽  
Comparative Genomic ◽  
Molecular Cytogenetic ◽  
Black And White ◽  
Chromosomal Changes ◽  
Molecular Cytogenetic Techniques

Classic cytogenetics has evolved from black and white to technicolor images of chromosomes as a result of advances in fluorescence in situ hybridization (FISH) techniques, and is now called molecular cytogenetics. Improvements in the quality and diversity of probes suitable for FISH, coupled with advances in computerized image analysis, now permit the genome or tissue of interest to be analyzed in detail on a glass slide. It is evident that the growing list of options for cytogenetic analysis has improved the understanding of chromosomal changes in disease initiation, progression, and response to treatment. The contributions of classic and molecular cytogenetics to the study of brain tumors have provided scientists and clinicians alike with new avenues for investigation. In this review the authors summarize the contributions of molecular cytogenetics to the study of brain tumors, encompassing the findings of classic cytogenetics, interphase- and metaphase-based FISH studies, spectral karyotyping, and metaphase- and array-based comparative genomic hybridization. In addition, this review also details the role of molecular cytogenetic techniques in other aspects of understanding the pathogenesis of brain tumors, including xenograft, cancer stem cell, and telomere length studies.

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