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Evolution of visual sensory system in the common barbel (Barbus barbus)
Konvičková, Zuzana ; Musilová, Zuzana (advisor) ; Pergner, Jiří (referee)
Vertebrates inherited five visual opsin classes from their common ancestor expressed in the rod and cone cells. While many vertebrate groups reduced the number of visual pigments they possess, opsins of teleost fishes have diversified. Today, teleosts outstand among all vertebrates in the number of visual pigments and photoreceptor types. This extraordinary development was also facilitated by the teleost- specific whole genome duplication that occurred approximately 350 Mya. Here I explored teleost species that recently underwent subsequent whole genome duplication - the common barbel (Barbus barbus). This event was accompanied by hybridization (i.e., it was allopolyploidization) and resulted in 14 opsin genes found in the genome, an unusually high number even among teleosts. In this thesis, I studied the effects of such an expanded opsin repertoire on the visual abilities of the common barbel. Based on the whole genome, I have completed the reference sequences of all opsin genes and revealed their positions on the chromosomes. I analysed retinal transcriptomes and constructed developmental series of opsin expression profiles. All opsin genes were preserved functional in the genome, even though one of them (green-sensitive, named RH2-4 in this thesis) was not found in the transcriptome and showed...
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Molecular genetic approaches used in preimplantation genetic analysis of human embryos in assisted reproduction
Limbergová, Jana ; Machoň, Ondřej (advisor) ; Šimoník, Ondřej (referee)
Preimplantation genetic testing is one of the main clinical procedures that take place in assisted reproduction centres around the world. It is sought after by couples for many different reasons. The causes of problems may relate to hereditary diseases that potential parents do not wish to pass on to their offspring, or may include the risk of transmission of chromosomal alterations. In addition, infertility may be caused by an increased risk of embryo aneuploidy, which correlates with mother's increased age and other factors that this bachelor thesis discusses. The bachelor thesis also focuses on up-to-date approaches to cytogenetic examinations for reproductive genetics and discusses recent clinical molecular methods that are used to improve the diagnosis and therapy of infertile couples. These are, for example, microarray methods, quantitative real-time PCR or next-generation sequencing and methods of whole-genome amplification. This work also summarizes methods that are gradually being abandoned, e.g. fluorescent in situ hybridization, and compares their advantages and disadvantages. Key words: preimplantation genetic testing, aneuploidy, structural rearrangements, monogenic diseases, biopsy, fluorescent in situ hybridization, microarray methods, comparative genomic hybridization, quantitative...
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Karyotype analysis of mygalomorph spiders of the families Hexathelidae and Dipluridae
Hrubá, Barbora ; Král, Jiří (advisor) ; Sember, Alexandr (referee)
Cytogenetics of spiders of the infraorder Mygalomorphae is largely unknown. My thesis is focused on the karyotype evolution of spiders of families Hexathelidae Dipluridae, which are basal groups of the superfamily Avicularioidea. In this thesis the evolution of diploid chromosome number, chromosome morfology, and also sex chromosomes are studied. The karyotypes of the mygalomorphs are quite interesting for their diversity. Many members of the family Hexathelidae exhibit extreme systems of a chromosomal sex determination which are composed by many gonosomes X. The meiotic division of males is analyzed. Some species exhibit modifications of this division such as achiazmatic meiosis. Selected markers have been detected by fluorescence in situ hybridization. Keywords: evolution, fluorescent in situ hybridization, karyotype, nucleolus organizer region, sex chromosomes, spider
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Analysis of chromosomal aberrations in sperm by fluorescence in situ hybridization
Bendová, Petra ; Diblík, Jan (advisor) ; Novotná, Drahuše (referee)
The presented bachelor work is focused on the determination of frequency chromosomally abnormal sperm in the semen of healthy men (donors) with normal karyotype (46, XY). The important process, which plays an irreplaceable role in the development of numerical aberrations of chromosomes or structural abnormalities in the segregation of the gametes, is meiosis. Therefore, I devote much attention on meiosis in the theoretical part. The theoretical part is focused on the process of pre mature sperm (spermatogenesis), and the consequences of fertilize the oocyte by aneuploid sperm. In my work I present an overview of numerical abnormalities in autosomes and gonosomes and their frequency and distribution of gametes in healthy men. I also focused on the distribution and a brief description of structural aberrations affecting chromosomes and not least I paid attention on method of multicolor interphase fluorescence in situ hybridization, which in combination with sperm chromatin dekondenzation become irreplaceable and valuable research tool for rapid analysis of chromosomal abnormalities in large sperm samples. The experimental part of bachelor work deals with monitoring the frequency of selected numerical abnormalities in sperm samples of five donors aged 23 to 30 years with the use of I-FISH (fluorescence in situ...
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Karyotype analysis of mesothelid spiders
Prokopcová, Lenka ; Král, Jiří (advisor) ; Zrzavá, Magda (referee)
Cytogenetics of mesothelid spiders is largely unkown. The presented diploma thesis is focused on the karyotype evolution of these spiders. As it is the most basal group of spiders, the analysis of its cytogenetics can bring important data about ancestral spider karyotype. In the framework of my thesis, I analysed diploid chromosome numbers, chromosome morphology, meiotic division, sex chromosomes and the pattern of selected molecular markers that were detected by fluorescence in situ hybridization. According to my results, mesothelid spiders have a high number of chromosomes and the prevalence of monoarmed chromosomes. Unlike other spiders, mesothelids have little differentiated sex chromosomes. Key words: evolution, spider, chromosome, karyotype, fluorescence in situ hybridization, nucleolar organiser region, sex chromosomes
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Analysis of sex chromosomes and gene clusters in selected mygalomorph and araneomorph spiders
Pappová, Michaela ; Král, Jiří (advisor) ; Johnson Pokorná, Martina (referee)
1 Abstract: The diploma thesis focuses on study of sex chromosomes evolution and repetitive organized genes of chosen mygalomorph and araneomorph spiders. Spiders are characterized by complexicity of sex chromosome systems, their karyotypes contain multiple sex chromosomes X. Besides multiple X chromosomes they also contain a pair or two pairs of nondiferentiated sex chromosomes X and Y. The used methods include methods of classical cytogenetics (preparation of chromosome slides, C-banding) and methods of molecular cytogenetics (fluorescent in situ hybridization and comparative genome hybridization). Complex sex systems were discovered in the studied Theraphosidae spiders. In Theraphosidae spiders Atropothele socotrana and Poecilotheria vittata neo-sex chromosomes were found. Analysis of molecular differentiation of sex chromosomes suggests low differentiation of Y chromosome in neo-sex chromosomes and pair of nondifferentiated sex chromosomes XY. In haplogyne spider Kukulcania aff. hibernalis (X1X2Y), the Y chromosome was significantly differentiated, male specific signal covered the whole chromosome. Detection of 18S rDNA showed that karyotypes of majority of analysed Theraphosidae spiders and haplogyne spiders contain low number (1 or 2) of nucleolar organizing regions localized terminally, which...
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Karyotype analysis of mesothelid spiders
Prokopcová, Lenka ; Král, Jiří (advisor) ; Zrzavá, Magda (referee)
Cytogenetics of mesothelid spiders is largely unkown. The presented diploma thesis is focused on the karyotype evolution of these spiders. As it is the most basal group of spiders, the analysis of its cytogenetics can bring important data about ancestral spider karyotype. In the framework of my thesis, I analysed diploid chromosome numbers, chromosome morphology, meiotic division, sex chromosomes and the pattern of selected molecular markers that were detected by fluorescence in situ hybridization. According to my results, mesothelid spiders have a high number of chromosomes and the prevalence of monoarmed chromosomes. Unlike other spiders, mesothelids have little differentiated sex chromosomes. Key words: evolution, spider, chromosome, karyotype, fluorescence in situ hybridization, nucleolar organiser region, sex chromosomes
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Karyotype analysis of mygalomorph spiders of the families Hexathelidae and Dipluridae
Hrubá, Barbora ; Král, Jiří (advisor) ; Sember, Alexandr (referee)
Cytogenetics of spiders of the infraorder Mygalomorphae is largely unknown. My thesis is focused on the karyotype evolution of spiders of families Hexathelidae Dipluridae, which are basal groups of the superfamily Avicularioidea. In this thesis the evolution of diploid chromosome number, chromosome morfology, and also sex chromosomes are studied. The karyotypes of the mygalomorphs are quite interesting for their diversity. Many members of the family Hexathelidae exhibit extreme systems of a chromosomal sex determination which are composed by many gonosomes X. The meiotic division of males is analyzed. Some species exhibit modifications of this division such as achiazmatic meiosis. Selected markers have been detected by fluorescence in situ hybridization. Keywords: evolution, fluorescent in situ hybridization, karyotype, nucleolus organizer region, sex chromosomes, spider
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Methods of molecular cytogenetics: evolution, analysis, comparison
Budková, Kateřina ; Šolc, Roman (advisor) ; Daňková, Pavlína (referee)
Cytogenetics is a branch of genetics that is concerned with the study of the structure and function of chromosomes, especially during the cell division. Chromosomal abnormalities that may occur during this process are often associated with the emergence of various diseases. Cytogenetic methods are primarily used to detect these abnormalities. Traditional methods allow resolution at the level of whole chromosomes, the introduction of new molecular methods led to a significant refinement of these analyzes. Today's methods allow easy analysis of chromosomes to the level of individual sequences. Despite significant differences between classical and modern cytogenetic techniques, classical cytogenetic methods are not evil, but they are still widely used. Combining classical and molecular methods in order to obtain a comprehensive analysis of chromosomal rearrangements is no exception.
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