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Sexual dimorphism in the mouse olfactory system
Kuntová, Barbora ; Stopka, Pavel (advisor) ; Havlíček, Jan (referee) ; Žídek, Lukáš (referee)
Sexually dimorphic behaviour of the house mouse (Mus musculus musculus) relies on various physical and chemical cues, however, chemical signals are the most essential cues for individual recognition and in causing various priming effects on reproductive behaviour of the receiver. House mice belong to macrosmatic mammals, and thus, their sense of smell is highly developed and is able to recognize a wide spectrum of ligands from other individuals and from their surrounding environment. Volatile signals belong to organic compounds that are produced by most tissues, and may have harmfull effects on cells, and thus they are transported out of the body with lipocalin transporters where some of them may function as signals. These volatile signals are able to stimulate chemosensory neuronal receptors, and thus, yield particular responses in neural circuits. The ligand sensing has a differential effect upon males and females, however, it has not been shown yet in wild mice whether these differences are also caused by the variation in receptors and neural processing, or rather by differential expression of signals typical for each sex. The aim of this thesis was to perform comparative analysis of orofacial mucosal tissues to determine the specificity of expression of particular lipocalins. For the first time...
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Chemical signals and reproductive processes of the house mouse (Mus musculus)
Černá, Martina ; Stopka, Pavel (advisor) ; Petr, Jaroslav (referee) ; Sedláček, Radislav (referee)
The aim of my thesis was to identify proteins involved in chemical communication and especially those that are involved in sexual signalling. Volatile chemical signals are transported with lipocalins in their beta-barrel structure to present their ligands to receptors or out of the body. Thus, I focused on the identification of these proteins in saliva and vaginal secretion of the house mouse using proteomic and transcriptomic approaches. Due to a cyclic manner of reproduction and its hormonal control, I have also focused on the role of estradiol on sperm phenotype in the laboratory mouse. We have identified an elevated sexual dimorphism in several lipocalins (i.e. 10 out of 20) in the saliva proteome where they may play a role in sexual signalling (i.e. similar to their described roles in the mouse urine). Interestingly, vaginal secretion also contains lipocalins and they rise from proestrus to estrus and remain steady during metestrus. Such variation provides evidence that they serve sexual signalling, however, due to their elevated levels during metestrus it is most likely that their ligands function as signals and not the proteins themselves. On the level of sperm phenotype, we have provided evidence, that experimental concentrations of estradiol have differential effects on sperm. This is due...
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Perception of individual variation in body odour in human adults
Fialová, Jitka ; Havlíček, Jan (advisor) ; Schaal, Benoist (referee) ; Stopka, Pavel (referee)
The thesis consists of two parts. The first part introduces the topic of human chemical communication and reviews current evidence on individual variation in human body odour and its perception. This part is framed by sexual selection theory. In the first chapter, the concept of the theory of communication is introduced followed by a discussion on the specifics of chemical communication. Next, the formation of individually specific body odour signatures with reference to skin glands, their volatile products and the subsequent metabolization by skin microflora is described. The next chapters are dedicated to selected interindividual body odour cues such as sex and kin recognition, genetic compatibility in genes of Major Histocompatibility Complex, and health and reproductive status in a mate choice context. Furthermore, interactions between perfumes and body odours are discussed. Finally, methods of body odour sampling are introduced and a rationale behind presenting individual samples or body odour blends is discussed. The second part is comprised of six scientific papers, specifically three reviews and three empirical studies. Review papers summarize factors affecting human body odour quality with emphasis on diet and affective states. The first text shows that human body odours contain cues to...
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Levels of physiologically active substances during the ontogeny of social hierarchy in two house mouse subspecies
Bufková Daniszová, Kristina ; Stopka, Pavel (advisor) ; Dušek, Adam (referee) ; Sedláček, František (referee)
2 Abstract Social interactions of animals are represented by a complex network of relations among individuals affected by many factors. From a practical point of view, it is very complicated or even impossible to observe these interactions under natural conditions and consider all fundamental underlying factors. A suitable solution may be, for example, to use a social environment of so called fraternal pairs. With a simplified model represented by pairs of brothers it is possible - given a properly set length of the experiment and age of the individuals involved in it - to reveal various aspects of establishing social hierarchies. A key factor influencing an individual's position (especially of a male) in the group is considered to be his physical fitness which is usually connected with his size and hence with his body mass. In addition to physical qualities of individuals, a fundamental role for achieving the "social" success is also played by the ability to communicate these qualities (e.g. through chemical signals) to other members of the social group. Apart from somatic and communication characteristics determining chances of an individual to succeed in the social competition, the internal regulation of these aspects presents another interesting subject for studies. In this respect a special attention...
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The role of caudal organ in chemical communication in field mice of the genus Apodemus
Obstová, Lucie ; Stopka, Pavel (advisor) ; Petrák, Jiří (referee)
This thesis deals with the chemical communication of wood mouse of genus Apodemus with a focus on the special sebaceous gland on the ventral side of the tail - the caudal organ. Role of the caudal organ in chemical communication is unclear. To illustrate the function of the sebaceous glands, I decided to use metods of 454 pyrosequencing, proteomic analysis and qPCR. I created a database of transcriptionally active genes of caudal organ using 454 pyrosequencing. In proteomic analysis, I focused on proteins with a molecular weight 14-18kDa to verify the presence of the lipocalins, proteins involved in chemical communication. I compared the relative mRNA abundance of selected genes between individuals of the opposite sex using qPCR. In my research, I found that the caudal organ is active organ and caudal organ could participate in chemical communication. I detected a different relative mRNA abundance of genes in males and females at the mRNA level, which could play a role in differences in development of caudal organ between individuals of the opposite sex. Finally, I suggested possible ways of using the caudal organ in the chemical communication in wood mouse genus Apodemus from the obtained results. Key words: caudal organ, Apodemus, chemical communication, FABP5, Scd1
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Molecular epidemiology of selected viral, bacterial and fungal disease of honeybees in the Czech Republic
Ryba, Štěpán ; Stopka, Pavel (advisor) ; Voříšek, Karel (referee) ; Půža, Vladimír (referee)
4 Summary Altogether, the six most common bee viruses which infect the honey bee (Apis mellifera) were monitored in the territory of the Czech Republic between 2006 and 2009. Parallel infections of viruses (DWV, ABPV and BQCV) in bee adults and parallel co- infection of viruses with fungal diseases caused by Nosema apis and Nosema ceranae were confirmed by PCR tests. A new sensitive method of detection of the originator of the American foulbrood (Paenibacillus larvae) from bee debris was developed for the practical use of detection of AFB disease in bee populations. Various approaches for the extraction of spores from bee debris and lyses of spores were compared. The sensitivity of PCR tests for the presence of Paenibacillus larvae in debris was compared with the classic cultivation method. The PCR method for the detection American foulbrood was further studied and developed to be more efficient. A new method, based on a matrix-like sample re-arrangement and a use of pooled samples, has been developed for testing 1000 samples in 35 PCR reactions. Another goal was to develop a robust and fast screening method for American foulbrood based on the cultivation test using paper sheets RIDA®COUNT with a specific cultivation medium, specific selection conditions for Paenibacillus larvae and chromogen visualization...
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Identification and modeling of gene expression regulatory networks during streptomycetes germination
Straková, Eva ; Vohradský, Jiří (advisor) ; Stopka, Pavel (referee) ; Vondrášek, Jiří (referee)
Streptomycetes have been studied mostly as producers of antibiotics and for fundamentals of complex bacterial cell development. Here, transcriptomic and proteomic approaches were applied to systems study of Streptomyces coelicolor germination as a developmental transition from dormancy to the vegetative stage. The time dynamics of the gene expression levels represented by mRNA and intracellular protein accumulation and synthesis were measured throughout 5.5 h of germination at 13 time points by employing both DNA microarray and two-dimensional gel electrophoresis techniques. Using a numerical model of gene expression, genetic networks were reconstructed and functional groups of genes controlled by the sigma factors were identified. Modeling of the regulatory interactions provided a set of parameters allowing simulate kinetics of gene expression control among the sigma factors and their target genes. Particularly regulons of two sigma factors, SigR and HrdD, were identified. The analysis assigned their key role during the germination process. Analysis of global trends in the gene/protein expression revealed that the full capability of regulatory mechanisms responding to the environmental cues is reached within the first hour of germination, and identified the basic gene/protein functional groups...
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Generation and analysis of mutant mouse models to study pathophysiological roles of KLK5 and KLK7 in epidermis
Kašpárek, Petr ; Sedláček, Radislav (advisor) ; Stopka, Pavel (referee) ; Machoň, Ondřej (referee)
Kallikrein-related peptidases (KLKs) constitute a family of closely related serine proteases encoded by genes clustered in one chromosomal locus. KLKs are widely expressed in a variety of tissues and numerous in vitro experiments suggest their important roles in many physiological and pathological processes. However, the biological roles of KLKs in vivo are often obscured mainly due to unavailability of suitable animal models. Although gene deficient mouse models were generated for several KLK genes, they had limited use for understanding the roles of individual proteases in the complex environment in vivo. One of the main obstacles which hampers in vivo analysis is partial functional overlap between some KLKs. This makes traditional single-gene deficient animal models an inadequate tool to address the biological impact of the gene deficiency as compensatory mechanisms often result in a lack of phenotype. In this work, we used the transcription activator-like effector nuclease (TALEN) technology to generate several novel mutant mouse models to study the complex KLK proteolytic pathways and their roles in healthy organism and in disease. We prepared a novel mouse model for Netherton syndrome (NS), an autosomal recessive skin disorder caused by mutation in the gene SPINK5, which encodes the KLK-inhibitor...
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