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Digital image analysis of mitotic chromosomes
Hávová, Mariana ; Babula, Petr (referee) ; Škutková, Helena (advisor)
Changes in chromosome number and structure may cause serious diseases. Cytogenetic tests leadin to set of karyotype are done for detecting these abnormalities. Chromosomes are visualised with proper methods and karyotype is made up most often. Manual karyotyping is time-consuming and expensive task. Because of this, researchers have been developing automated karyotyping systems. Karyotyping systems classify chromosomes into classes based on their characteristic features. Overlapping and bent chromosomes are limitations for automatic classification since they ocur at almost every mitosis. Accuracy and reliability of karyotyping systems still depend on the human intervention. Overcoming of these problems and development of fully automated system is the aim of modern approaches.
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Karyotype evolution of selected families of entelegyne spiders
Kotz, Matěj ; Král, Jiří (advisor) ; Nguyen, Petr (referee)
The Araneoidea superfamily is a diverse clade of spiders with a great species diversity. The whole superfamily displays considerable conservativeness of observed karyotypes. Most likely ancestral karyotype in males is 24 acrocentric chromosomes with X1X2 sex determination system. The goal of this study is to explore the karyotype diversity of two araneoid families - Araneidae and Mimetidae. The majority of studied species exhibit the ancestral karyotype. In some species of the aformentioned families was observed sudden increase in chromosome numbers, up to 2n♂ = 52 in Araneidae and up to 2n♂ = 57 in Mimetidae. The latter number is the highest chromosome count observed in Entelegynae so far. Increase in 2n goes hand in hand with increase in sex chromosome numbers, leading up to X1X2X3X40 system in Araneidae and up to X1X2X3X4X5X6X70 in Mimetidae. I suggest polyploidy as a possible mechanism of the increase. To test this hypothesis, I measured the size of the genome using flow cytometry and used fluorescence in situ hybridization for the detection of 18S rRNA and 5S rRNA genes. For one species, probe for U2 snRNA gene was also optimized as part of this thesis. In many species studied, these techniques were used for the first time ever. In the case of the family Mimetidae, the largest genomes in...
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Karyotype evolution in estrildid finches of the genus Lonchura
Janáková, Šárka ; Reifová, Radka (advisor) ; Augstenová, Barbora (referee)
The karyotype, the set of all chromosomes in a cell, is an important characteristics of individual species of both animals and plants. The karyotype of birds is typically stable in terms of the number of chromosomes and their size, and rearrangements between different chromosomes are rare. The changes that occur between the karyotypes of avian species are more often intrachromosomal, such as inversions or duplications and deletions. It is still unclear how often such changes occur between closely related species and whether they can contribute to speciation. This work aims to compare the karyotypes of a total of five closely related species of songbirds of the genus Lonchura (munias and mannikins). Comparing chromosomal changes in such recently diverged species allows us to observe changes that might contribute to speciation. The study is based on karyotype analysis by immunofluorescence staining of synapsed pachytene chromosomes and subsequent identification and comparison of chromosome types. This method allows determination of the changes that are manifested by pericentric inversions where the position of the centromere is changed. The results confirm the general idea of avian karyotypes as very conservative in terms of chromosome number and their sizes. However, in some chromosomes we observed...
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Mechanisms of reproductive isolation between two hybridizing passerine species, the common nightingale and the thrush nightingale
Poignet, Manon ; Reifová, Radka (advisor) ; Choleva, Lukáš (referee) ; Kratochvíl, Lukáš (referee)
iv - ABSTRACT (in English) - A key step in the origin of new species is the evolution of the reproductive isolation mechanisms separating the gene pools of newly emerging species. Despite a growing number of studies, we still know very little about the molecular, physiological and genetic mechanisms underlying reproductive isolation. Hybrid zones represent useful examples of ongoing species differentiation and can provide information about the nature of reproductive isolation separating the species and mechanisms facilitating or limiting the hybridization. Using two naturally hybridizing passerine species, the common nightingale (Luscinia megarhynchos) and the thrush nightingale (L. luscinia), we evaluated several mechanisms of reproductive isolation at the chromosomal, phenotypic, and genetic levels, with a focus on postmating prezygotic (PMPZ) and postzygotic isolation mechanisms. These nightingale species diverged ~1.8 Mya and currently hybridize in a secondary contact zone running across Central and Eastern Europe. First, we analysed the genetic composition of the secondary contact zone, demonstrating that parental forms are predominant in the hybrid zone, with few F1 hybrids (3.4%), early backcross hybrids (3.1%) and the absence of F2 hybrids. This suggests strong but incomplete reproductive isolation....
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Evolution of karyotype and sex chromosomes in African and American clades of theraphosid mygalomorph spiders
Turečková, Eva ; Forman, Martin (advisor) ; Šťáhlavský, František (referee)
The family Theraphosidae (tarantulas) are generally known of spiders, often keept as hobby pets. Despite their popularity, many aspect of tarantula biology are still omitted. This include also cytogenetic research. In order, to fill this gap I analysed chromosomal constitution in 13 species of tarantulas belonging to the different genera from Africa and south/central America. Data set in this thesis included, also, the tarantula with so far the best covered genome Acanthoscuria genicullata, and the iconic, giant spider Theraphosa stirmi. Conventional giemsa staining of male meiotic plates was accompanied with visualisation of major rRNA clusters using fluorescence in situ hybridization with 18S rDNA probe. Diploid counts ranges considerably from 31 (Ceratogyrus meridionalis) to 83 (Theraphosa stirmi). In some specimens presence of multivalents indicated a possibility of hybridization of different chromosomal races in a hobby lineages. Various types of differentiated sex chromosomes have been proven, including X0, X1X20 X1X2X30 systems. Interestingly, the diversity in karyotype features was not linked with diversity in major rDNA cluster number. Most of the species displayed one prominent locus located on autosomes. Two species exhibited polymorphism in the presence of one more additive smaller...
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Evolution of karyotypes and sex determination in the turtle family Geoemydidae
Clemente, Lorenzo ; Rovatsos, Michail (advisor) ; Montiel Jimenez, Eugenia Elisabet (referee) ; Castiglia, Riccardo (referee)
(IN ENGLISH) The majority of studied turtles show temperature-dependent sex determination, but genotypic sex determination (i.e. presence of sex chromosomes) was identified sporadically. This thesis aims to investigate and expand our knowledge on the evolution of the karyotype and the sex determination in turtles, particularly focusing on the family Geoemydidae, a group of turtles with previously documented variability in sex determination systems. The presence of sex chromosomes was explored by a combination of conventional and molecular cytogenetic techniques for the analysis of karyotypes, distribution of constitutive heterochromatin (C-banding) and repetitive elements and comparative genome hybridization (FISH, CGH). In total, 49 species of turtles from nine different families were cytogenetically examined in this study. In the family Geoemydidae, a remarkable similarity in karyotypes was identified, consisting of 2n=52 chromosomes (which is suggested to be the ancestral diploid number for all turtles) and a similar topology of rDNA loci and telomeric repeats. Sharma et al. (1975) previously reported ZZ/ZW sex chromosomes in Pangshura smithii. However, in the analysis presented in this thesis, it is suggested a possible misidentification of these sex chromosomes due to erroneous pairing of...
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Cytogenetics of bed bugs (Cimicidae) as a model representative of true bugs (Heteroptera)
Sadílek, David ; Vilímová, Jitka (advisor) ; Johnson Pokorná, Martina (referee)
Cytogenetics of bed bugs (Cimicidae) as a model true bugs (Insecta: Heteroptera) The thesis provides current opinions about a phylogeny of bed bugs, family Cimicidae, and their classification within the order Heteroptera. There are briefly summarized cytological data about the order Heteroptera, known karyotypes of the cimicid subfamilies and introduction to cytogenetics of species Cimex lectularius Linnaeus, 1758. Heteroptera species differ from other organisms by holokinetic chromosomes, a character, in which quite frequently take place chromosomal fragmentation and fusion. Several families posses smaller pair of m chromosomes, which behave during meiosis differently from the other chromosomes. Diploid chromosomal number is extremely variable, it ranges from four (family Belostomatidae) to 80 chromosomes (family Miridae). The behavior of chromosomes in the family Cimicidae is very specific. Sex-chromosomes univalents are connected by thread-like collochores and form so-called pseudobivalents during achiasmatic post-reductional meiosis. Rare polymorphism in a number of X chromosomes occurs in Cimex lectularius populations. The diploid karyotype always consists of 26 autosomal chromosomes and at least two but up to 15 X chromosomes and one Y chromosome. The number of X chromosomes can be variable...
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Karyotype evolution of African clades of theraphosid mygalomorphs
Košátko, Prokop ; Král, Jiří (advisor) ; Nguyen, Petr (referee)
Karyotypes of mygalomorph spiders are not satisfactorily known. This thesis is focused on the basic cytogenetic analysis of selected species of African clades of theraphosid mygalomorphs. It includes four subfamilies: Eumenophorinae, Harpactirinae, Ischnocolinae and Stromatopelminae. Diploid numbers, chromosome morphology, sex chromosome systems and chromosome behaviour in male germline in the selected species of African theraphosid subfamilies were studied. The findings support published results, that refer of high karyotype diversity in Theraphosidae. Diploid chromosome number reduction is probably a basic trend of theraphosid karyotype evolution. The majority of analysed species exhibited one, two or three sex chromosomes. In some species neo-sex chromosome systems were found. In some species one or two sex chromosome pairs (SCP), composed of chromosomes which lack morphological differentiation were detected. Nucleolus organizer regions were detected by fluorescent in situ hybridization in several species. Constitutive heterochromatin detection was performed by C-banding in two species. Keywords: constitutive heterochromatin, diploid number, karyotype, fluorescence in situ hybridization, Mygalomorphae, nucleolus organizer region, SCP, sex chromosome, spider, Theraphosidae
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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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Evolution of sex chromosomes and karyotypes in iguanids (Squamata: Iguanidae sensu lato)
Altmanová, Marie ; Kratochvíl, Lukáš (advisor) ; Symonová, Radka (referee)
Iguanids are greatly diversed group of squamate reptiles (Reptilia, Squamata). Although there were many phylogenetical studies made, the relationships among main lineages are still unclear. In cytogenetical point of view is this group relatively conservative. In three species-rich genus only, the diversity of karyotypes is relatively considerable: Anolis, Liolaemus a Sceloporus. Diploid chromosome number varies between 19 and 48. Ancestral karyotype is defined as 2n=36, with 12 macrochromosomes and 24 microchromosomes. Morphologically the macrochromosomes are mostly metacentric, rarely acrocentric or telocentric. In karyotype evolution Robertsonian rearrangements dominate apparently, where number of chromosome arms is preserved. Interspecific variability or variability between populations were observed. Sex seems to be determined by genotype for whole group (GSD). Across the group probably attended conditionally to multiple differentiation of sex chromosomes. In 75 of 219 caryotyped were detected XX/XY type of sex chromosomes or derived multiple sex system X 1X 1X 2X 2/X 1X 2Y. The transition from multiple sex chromosomes back to XX/XY type was observed in one species. Chromosome Y shows morphological variability in size and shape, from microchromosom to huge metacentric macrochromosom. Homology of sex...
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