Evolution of chemical signals, olfaction and nasal immunity in muroid rodents
| Název práce v češtině: | Evoluce chemických signálů, olfakce a nosní imunity u myšovitých hlodavců |
|---|---|
| Název v anglickém jazyce: | Evolution of chemical signals, olfaction and nasal immunity in muroid rodents |
| Klíčová slova: | čich, chemické signály, feromony, hlodavci |
| Klíčová slova anglicky: | olfaction, chemical signals, pheromones, rodents |
| Akademický rok vypsání: | 2021/2022 |
| Typ práce: | disertační práce |
| Jazyk práce: | angličtina |
| Ústav: | Katedra zoologie (31-170) |
| Vedoucí / školitel: | prof. Mgr. Pavel Stopka, Ph.D. |
| Řešitel: | skrytý - zadáno a potvrzeno stud. odd. |
| Datum přihlášení: | 15.10.2021 |
| Datum zadání: | 15.10.2021 |
| Datum potvrzení stud. oddělením: | 15.10.2021 |
| Konzultanti: | Mgr. Romana Stopková, Ph.D. |
| Mgr. Klára Hlouchová, Ph.D. | |
| Zásady pro vypracování |
| Schneiderová I.: Obratlovci střední Evropy
Starostová et al.: Speciální exkurze ze zoologie |
| Seznam odborné literatury |
| 1. Thoß, M. et al. Regulation of volatile and non-volatile pheromone attractants depends upon male social status. Sci. Rep.9, 1–14 (2019).
2. Gonzalez, S. et al. Conceptual aspects of self and nonself discrimination. Self/Nonself - Immune Recognit. Signal.2, 19–25 (2011). 3. Kuntová, B., Stopková, R. & Stopka, P. Transcriptomic and proteomic profiling revealed high proportions of odorant binding and antimicrobial defense proteins in olfactory tissues of the house mouse. Front. Genet.9, 1–13 (2018). 4. Stopkova, R., Klempt, P., Kuntova, B. & Stopka, P. On the tear proteome of the house mouse (Mus musculus musculus) in relation to chemical signalling. PeerJ2017, (2017). 5. Tew, T. E. & Macdonald, D. W. Dynamics of space use and male vigour amongst wood mice, Apodemus sylvaticus, in the cereal ecosystem. Behav. Ecol. Sociobiol.34, 337–345 (1994). 6. Raulo, A. et al. Social networks strongly predict the gut microbiota of wild mice. ISME J. (2021). doi:10.1038/s41396-021-00949-3 7. Tretyachenko, V., Voráček, V., Souček, R., Fujishima, K. & Hlouchová, K. CoLiDe: Combinatorial Library Design tool for probing protein sequence space. Bioinformatics (2020). doi:10.1093/bioinformatics/btaa804 |
| Předběžná náplň práce |
| a) Metabolomics as a tool to study odours in phylogenetic contexts
Advances in the development of new OMICSs techniques, and the sequence of the mouse and other genomes brings ever new knowledge about all RNAs and proteins in the laboratory and wild rodents and help to understand of life in the context of its environment from level of organic compounds, peptides, microbes, individuals, populations and communities. Earlier, most studies using these techniques bring only limited informations about few selected organic compounds and classical biological topics like evolution and function was studied mainly in perspective of methods DNA, RNA and protein sequencing. In addition, metabolomics, is the next level in genetics as it provides the profiles of organic compounds typical for particular environments, microbes, microbiota and their hosts, including the body odours and volatile pheromones 1. Thus, one of the main aims of this project is to extract metabolomic signatures (odorous substances in saliva, urine, faeces) from small mammalian species living in different environments in central Europe to see whether these signatures rather reflect their phylogeny (similarly as DNA-based branching), microbiota or their ecology (incl. samples from environment, food, etc.) and behaviour (territorial, social). b) Nostrils as a filter to environmental immune challenges The nostrils face permanent contact with outside enviroment and except sensoric function of olfaction they represent also the first filter of immune system. This system of filtering thus provides immediate defence against pathogens, based on receptors, which recognize nonself molecules present in pathogens, but not present in the host and the host microbiota2. In addition to the innate immune receptors, there is a system of proteins (e.g. lipocalins) continuously removing the organic content from defeated bacteria as well as potentially toxic organic compounds and odorants that enter the nostrils during ventilation. Detailed characterization of this system will be provided within this project using the proteomic analysis of nasal lavages, and major morphological structures (VNO, MOE, NALT) in two wild species of rodents with different mating systems (promiscuous Apodemus sylvaticus, monogamous A. uralensis) similarly as we studied in the house mouse3. c) Nasal lipocalins Another aim is to determine whether lipocalins from the family of odorant binding proteins (OBP1-8) have direct antimicrobial effects in addition to the ability to bind and transport VOCs. We assume that this effect is fortified by the amphipathic distribution of amino-acid residua of mouse OBPs where positively charged side of the protein has the potential to destroy negatively charged bacterial membranes4. In this context, we ask an important question. Is this dual system of olfaction and immunity something that specifically evolved in the house mouse for its striking dependences on olfactory cues, or is it a system that can be detected in varying stages of evolution also in other mammalian species? This is why we aim to do the analysis of nasal proteomes in small rodents differing in their ecology, distribution patterns, and their visual abilities (e.g. larger eyes in Apodemus, smaller eyes in voles, Microtus). d) Sociality as a driver for microbial diversity One of the most important aspects of social life in mammals is that different individuals and even different species have varying frequencies of meeting each other. This behavior can be good predictor of microbiota composition and dynamics transfer pathogens as well. In this project, we aim to use 16S- RNA sequencing to extract the microbial signatures for testing the hypothesis that close social contacts generate similarities between individuals in social networks and potentially also in different species in mixed populations5,6. e) Synthetic peptides as a tool to simulate potential immune challenges We also ask whether the nasal capacity to detect odours and pathogens is correlated and whether it is environmentally modulated, i.e. dynamically changing in different environments and/or due to different composition of potential immune challenges. To solve the latter, we are now able to generate short synthetic peptide libraries of different sequences, including those of different viral and bacterial proteins as well as of random compositions. With these peptides we aim to simulate the inner rodent world (self) and a protein space that is completely unknown to the study subject, thus representing nonself types of peptides7. Presenting these peptides to the mouse nostrils in a set of experiments using the mass spectrometry analyses, consequent nasal lavages will provide us with the proteomic signatures, which aid to reveal particular proteins and biochemical pathways that are elevated due to the self and nonself peptide stimulation. |
| Předběžná náplň práce v anglickém jazyce |
| a) Metabolomics as a tool to study odours in phylogenetic contexts
Advances in the development of new OMICSs techniques, and the sequence of the mouse and other genomes brings ever new knowledge about all RNAs and proteins in the laboratory and wild rodents and help to understand of life in the context of its environment from level of organic compounds, peptides, microbes, individuals, populations and communities. Earlier, most studies using these techniques bring only limited informations about few selected organic compounds and classical biological topics like evolution and function was studied mainly in perspective of methods DNA, RNA and protein sequencing. In addition, metabolomics, is the next level in genetics as it provides the profiles of organic compounds typical for particular environments, microbes, microbiota and their hosts, including the body odours and volatile pheromones 1. Thus, one of the main aims of this project is to extract metabolomic signatures (odorous substances in saliva, urine, faeces) from small mammalian species living in different environments in central Europe to see whether these signatures rather reflect their phylogeny (similarly as DNA-based branching), microbiota or their ecology (incl. samples from environment, food, etc.) and behaviour (territorial, social). b) Nostrils as a filter to environmental immune challenges The nostrils face permanent contact with outside enviroment and except sensoric function of olfaction they represent also the first filter of immune system. This system of filtering thus provides immediate defence against pathogens, based on receptors, which recognize nonself molecules present in pathogens, but not present in the host and the host microbiota2. In addition to the innate immune receptors, there is a system of proteins (e.g. lipocalins) continuously removing the organic content from defeated bacteria as well as potentially toxic organic compounds and odorants that enter the nostrils during ventilation. Detailed characterization of this system will be provided within this project using the proteomic analysis of nasal lavages, and major morphological structures (VNO, MOE, NALT) in two wild species of rodents with different mating systems (promiscuous Apodemus sylvaticus, monogamous A. uralensis) similarly as we studied in the house mouse3. c) Nasal lipocalins Another aim is to determine whether lipocalins from the family of odorant binding proteins (OBP1-8) have direct antimicrobial effects in addition to the ability to bind and transport VOCs. We assume that this effect is fortified by the amphipathic distribution of amino-acid residua of mouse OBPs where positively charged side of the protein has the potential to destroy negatively charged bacterial membranes4. In this context, we ask an important question. Is this dual system of olfaction and immunity something that specifically evolved in the house mouse for its striking dependences on olfactory cues, or is it a system that can be detected in varying stages of evolution also in other mammalian species? This is why we aim to do the analysis of nasal proteomes in small rodents differing in their ecology, distribution patterns, and their visual abilities (e.g. larger eyes in Apodemus, smaller eyes in voles, Microtus). d) Sociality as a driver for microbial diversity One of the most important aspects of social life in mammals is that different individuals and even different species have varying frequencies of meeting each other. This behavior can be good predictor of microbiota composition and dynamics transfer pathogens as well. In this project, we aim to use 16S- RNA sequencing to extract the microbial signatures for testing the hypothesis that close social contacts generate similarities between individuals in social networks and potentially also in different species in mixed populations5,6. e) Synthetic peptides as a tool to simulate potential immune challenges We also ask whether the nasal capacity to detect odours and pathogens is correlated and whether it is environmentally modulated, i.e. dynamically changing in different environments and/or due to different composition of potential immune challenges. To solve the latter, we are now able to generate short synthetic peptide libraries of different sequences, including those of different viral and bacterial proteins as well as of random compositions. With these peptides we aim to simulate the inner rodent world (self) and a protein space that is completely unknown to the study subject, thus representing nonself types of peptides7. Presenting these peptides to the mouse nostrils in a set of experiments using the mass spectrometry analyses, consequent nasal lavages will provide us with the proteomic signatures, which aid to reveal particular proteins and biochemical pathways that are elevated due to the self and nonself peptide stimulation. |