GA ČR
GA ČR
Basic information: During mouse preimplantation development G1 is long and G2 short in the zygote, whereas G1 is extremely short and G2 very long in the 2-cell embryo. In somatic cells checkpoint kinase 1 (CHK1) and its substrate CDC25A phosphatase play a critical role in cell cycle checkpoints to preserve genome integrity. Mouse germ line knock-outs for Chk1 and Cdc25A are embryonically lethal due to post-implantation lethality but preimplantation development is normal, likely driven by maternal CHK1 and CDC25A protein. The function of the CHK1-CDC25A pathway during mammalian preimplantation development is not known. In a preliminary study we found that CHK1 is essential for checkpoint signaling in 2-cell embryos but surprisingly not in zygotes that do require CDC25A for mitosis. Using oocyte specific knock-outs of Chk1 and Cdc25A, combined with live-cell imaging techniques, we will determine the basis for differences in signaling and function of checkpoints between zygote and 2-cell embryos that contribute to the differences in cell cycle progression between zygotes and 2-cell embryos.
Registration No: 20-27742S
Project duration: 1. 1. 2020 – 31. 12. 2022, extended to 30. 6. 2023
Principal Investigator: RNDr. Petr Šolc, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Variability in environmentally relevant traits is key to the ability of species to adapt to changes in climatic and environmental conditions. It is therefore important to understand the determinants of such adaptive variation. We propose to sequence whole genomes of a large number of bank voles sampled along a north-south cline in genotypes and physiological phenotypes in Great Britain, spanning a wide gradient of altitude and climate. We will scan the genomes for regions of elevated geographic and genomic divergence in order to detect candidate loci involved in local adaptation and will test for gene-environment associations to identify the most important environmental factors driving adaptive divergence. Determining the mechanisms of local adaptation to environmental gradients in the bank vole will provide general insights relevant for adaptation of species to the current and future climate change.
Registration No: 20-11058S
Project duration: 1. 1. 2020 – 31. 12. 2022, extended to 30. 6. 2023
Principal Investigator: RNDr. Petr Kotlík, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Microbial metabolites and dietary factors influencing genome and epigenome in colorectal cancer development
Basic information: Gut microbiome and intestinal diseases, such as inflammatory bowel diseases and colorectal cancer, are tightly connected and, in association with the stage of the disease, the composition and function of microbes significantly differ. Another factor shaping the gut microbiome is a diet affecting the spectrum of luminal metabolites. These changes can influence cellular turnover and thus possible tumor development via genome or epigenome modifications of the host cells. In this proposal, we focus on microbial metabolome in experimental mouse models of gut inflammation and colorectal cancer after dietary interventions and subsequent analysis inflammation, oxidative stress, and genome and epigenome modifications during the disease induction and progress. In addition, we will study the metabolites in vitro targeting cellular pathways associated with inflammation, oxidative stress, epigenetic modifications and DNA damage repair processes. Simultaneously, we will collect and process fecal, mucosal and serum samples from respective patients to further validate experimental data.
Registration No: 20-03997S
Project duration: 1. 1. 2020 – 31. 12. 2022, extended to 30. 6. 2023
Principal Investigator: Mgr. Klára Kostovčíková, Ph.D. Institute of Microbiology CAS
Other solver: MUDr. Pavel Vodička, CSc. Institute of Experimental Medicine CAS
Other solver: Ing. Jiří Killer, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Novel functions of c-Myb during intramembranous bone formation: analyzing molecular interactions in craniofacial morphogenesis
Basic information: The c-Myb transcription factor is associated with maintaining the undifferentiated state of hematopoietic cells, controlling cell cycle and apoptosis. The recent studies pointed to its important role during process of bone formation. The proposed project aims to unravel the mechanisms of the newly described role of c-Myb in osteogenesis especially in the intramembranous type of ossification. First, the molecular modulation will be carried out in vitro. The functional experiments will be performed at osteoblasts cell line. The changes in expression of osteogenic genes will be analyzed using PCR Array technique. Second part includes structural description and localisation of c-Myb expression in vivo. The research of evolutionary aspect of c-Myb protein expression in will enrich the study. The osteogenic potential of c-Myb can be used in modern medical applications, e.g. for treatment of bone mass deficiency in tooth implantology.
Registration No: 19-15272Y
Project duration: 1. 1. 2019 – 31. 12. 2022, extended to 30. 6. 2023
Principal Investigator: Mgr. Veronika Oralová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Inhalation is the main route for exposure of organism to nanoparticles. Inhaled nanoparticles penetrate into the lung, translocate across the alveolar-capillary barrier into blood and they are transported to the secondary target organs. This project focuses on the influence of inhaled water-soluble and semi-soluble compounds of lead and cadmium nanoparticles found in the urban aerosol on target organs of mice and mechanisms of the metal nanoparticle clearance from different exposed tissues. Mice will be exposed to nanoparticles in inhalation chambers to provide physiological exposure similar to real air conditions in urban areas. The results will deepen the knowledge of the effects of inhaled metal nanoparticles with different physicochemical properties on individual target organs and will contribute to uncover possible tissue-specific responses to nanoparticles exposure. Determined molecular and cellular mechanisms of nanoparticle clearance from exposed organs will help to find new approaches how to enhance the nanoparticle clearance in polluted industrial areas.
Registration No: 20-02203S
Project duration: 1. 1. 2020 – 31. 12. 2022
Principal Investigator: RNDr. Pavel Mikuška, CSc. Institute of Analytical Chemistry of the CAS, v. v. i. (IAC)
Other solver: doc. RNDr. Marcela Buchtová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Other solver: Mgr. Tomáš Vaculovič, Ph.D. Masaryk University, Faculty of Science
Programmed DNA elimination: on the functions and mechanisms regulating the process in animal genomes with hemiclonal heredity
Basic information: The stability of the genome is essential and paramount to organisms for maintaining the precisely tuned functionality of their genomes. However, diverse eukaryotes carry out programmed DNA elimination in which portions or entire chromosomes are lost during individual development. In most cases, programmed DNA elimination is associated with either differentiation of somatic cells or sex determination. How different genomic segments are reproducibly retained or discarded is unknown. The proposed project intends to analyze molecular, chromosomal and genomic mechanisms to understand processes how whole genomes will be lost during programmed DNA elimination. From the comparative approach with included one group of fish (Hypseleotris) and one group of tetrapods (Pelophylax) with a hemiclonal heredity (hybridogenesis) we expect fundamental insights into eliminating processes during gametogenesis. Understanding the mechanisms by which vertebrates regulate such extensive remodelling of its genome will provide insights into factors that can promote stability and change in vertebrate genomes.
Registration No: 19-24559S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: RNDr. Lukáš Choleva, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: In the present project we intend to address the evolution of the freshwater fauna in Eurasia on very large geographic scale (continental-wide) and time scale (whole Cenozoic). The project shall reveal the major events that have shaped the freshwater fauna in Eurasia as well as the main geological and climatic events that have been responsible for these events. As model organism we will use freshwater fishes that occur in every river in Eurasia and have been present since the beginning of Cenozoic (family Nemacheilidae, order Cypriniformes). The methodology combines analyses of a large amount of samples by Sanger sequencing with an analysis of selected samples for a large amount of data (by next-generation sequencing). To our knowledge, this is the first approach to address the evolutionary history of freshwater animals across this geologically highly active region of Eurasia for such long geologic period. The study aims to provide a model study for further research on the evolutionary history of freshwater fauna throughout Eurasia.
Registration No: 19-18453S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Mgr. Vendula Bohlen-Šlechtová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
The relationship between cell size and the size of cellular organelles during early embryonic development in mammals
Basic information: Despite current progress in cell biology the mechanism how cells maintain their size still remain unresolved. Cellular volume has impact on the size of the organelles and a general architecture of tissues and organs and multiple pathological conditions are characterized by changes in cell sizes. A maintenance of cell volume presumably results from a balance between synthesis and destruction of cellular content, and whereas we have some knowledge about these two processes we do not understand how cells are actually measuring their volume. Preliminary hypotheses suppose adjustment of cell volume to unknown molecular ruler, cell ploidy or metabolic activity. None of the mechanisms proposed so far is however able to explain remarkable size differences of cells in tissues or maintenance of cell volume in non-proliferating cells. We propose to use scalable system of embryonic blastomeres to study the regulation and consequences of cell size changes. Our results should elucidate the relationship of the cell size and cell cycle progression, DNA content or metabolic activity of cells.
Registration No: 19-24528S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: MVDr. Martin Anger, CSc. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Craniofacial development includes intramembranous osteogenesis, where osteoblasts differentiate from mesenchymal precursors. Caspases are known to contribute to inflammation and cell death, however, recent evidence points to much broader spectrum of their functions. The latest data indicate the presence of active caspases in non-apoptotic cells of craniofacial bones in vivo. Additionally, pharmacological inhibition of caspase activity in osteoblastic cells derived from intramembranous bone resulted in decreased expression of osteocalcin, the key marker of osteoblasts´ differentiation and function. The proposed project aims to further clarify this novel osteogenic potential of caspases with focus to investigate the mechanism of osteogenic caspase action in bone cells using combination of pharmacological/genetic approaches and large scale transcriptomic and proteomic analyses. The project belongs to basic research with potential further applications particularly in molecular strategies for bone repair and regeneration.
Registration No: 19-14727S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: doc. Mgr. Petr Beneš, Ph.D. Masaryk University, Faculty of Science
Spoluřešitel: Prof. RNDr. Eva Matalová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Cytogenomics of African annual killifish species pairs: a unique model of early vertebrate sex chromosome evolution
Basic information: African annual killifishes of the genus Nothobranchius (Teleostei: Nothobranchiidae) are known for their unique adaptation to freshwater temporary water pools appearing during monsoons and desiccating during dry season. Diapausing embryos, fast development and short lifespan make them vital models for various biological disciplines. Sequenced genome of N. furzeri and current cytogenetic data suggest accelerated rate of karyotype and sex chromosome evolution in non-overlapping generations and small-sized populations, driven by genetic drift and allopatric mode of speciation. Given the striking variety of sex chromosome systems found in fishes so far, their relatively young age and rapid dynamics, killifishes offer the striking possibility to capture different phases of early vertebrate sex chromosome evolution and master sex-determining genes, with possibility to compare species complexes and sympatric taxons. Therefore, we aim to explore (not only) the Southern clade of Nothobranchius killifishes (seven species) using battery of modern cytogenomic and transcriptomic methods.
Registration No: 19-22346Y
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Mgr. Alexandr Sember, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Genome editing to treat Stargardts disease – Generation and phenotyping of a porcine model and development of a treatment approach
Basic information: Minipig represents unique, biomedical model thanks to its size and comparable physiological parameters with a man. Also the retina architecture is optimized for daily vision with the high concentration of cones in both species. Therefore we decided to create the transgenic minipig model of inherited retinal dystrophy – Stargardt disease type 1, caused by mutation in ABCA4 gene. The proposed ABCA4 gene editing, using CRISPR/Cas9 technology, will produce null mutation (V1973X) in one-cell minipig embryos. These embryos will be transferred in oviducts of the synchronized recipients. Piglets will be genotyped and the phenotype development will be followed by non-invasive (OCR, ERG, behavioral tests) and invasive tests (EM, IHC, WB, qPCR). For treatment of the transgenic minipigs, line ABCA4 V1973X, will be prepared vectors, first for in vitro testing, and subsequently for subretinal application in 6 month old minipigs. We suppose a sufficient amount of photoreceptors at this age to discover a positive effect of genome editing.
Registration No: 19-09628J
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Prof. MVDr. Jan Motlík, DrSc. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Major urinary proteins (MUPs) transport and protect volatile ligands but they can themselves act as olfactory cues. Since MUPs contribute to complex information about the donor they can play an important role in divergence between nascent species. Despite of being intensively studied, majority of the studies focused on the “classical” inbred strain C57 while wild house mouse populations have scarcely been studied. Here we focus on the following questions: What is the DNA sequence structure of the Mup cluster in 8 wild derived strains of 3 subspecies (musculus, domesticus, castaneus) and 4 non-commensal mouse species? How does copy number variation (CNV) contribute to the variation? Which genes are expressed on the RNA and protein levels? How do different types of social interactions affect expression of individual MUPs? For solving these questions we use modern genomic (long-reads sequencing, NGS, CNV identification with NGS and ddPCR), transcriptomic (qPCR of cDNA, NGS-based techniques), and proteomic (mass spectroscopy) methods.
Registration No: 19-19056S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: prof. RNDr. Miloš Macholán, CSc. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Huntington’s disease is inherited incurable neurodegenerative disease caused by mutation in gene for protein huntingtin. Emerging therapies are aimed at lowering of mutant huntingtin production to postpone the disease onset. Development of such disease-modifying treatments is complicated by lack of biomarkers that could be used to monitor therapeutic effects, mainly in the presymptomatic phase. Extracellular vesicles (EVs), mainly exosomes, are membraneous vesicles released by cells to body fluids. EV content and presence of huntingtin or its fragments may reflect changes in inaccessible central nervous system. The aim of this project is to map the protein composition of EVs in cerebrospinal fluid and blood plasma in porcine model of Huntington’s disease and to compare levels of selected proteins with EVs isolated from blood plasma of patients and healthy controls. We expect that the peripheral EVs could help us understand changes in central nervous system caused by the presence of mutant huntingtin. We also expect identification of candidate markers of the disease.
Registration No: 19-01747S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Mgr. Helena Kupcová Skalníková, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Other solver: Doc. MUDr. Jiří Klempíř, Ph.D. First Faculty of Medicine, Charles University
Basic information: Regulated mRNA translation is vital for germ cells, as well as for preimplantation embryos to produce new proteins in the spatial and temporal patterns that drive gamete and embryo development. With the increasing use of assisted reproduction techniques both in the field of human and farm animal reproduction, occurring during the last few decades, the important question arises of how the physiology of the “in vitro” produced embryos matches that of the embryos developed in vivo and “naturally”. This project is focused on the study of the differences in the temporal pattern of translation of specific transcripts between the MII oocytes matured in vitro and in vivo, as well as between the zygotes obtained after in vivo fertilization versus those obtained after in vitro fertilization. Furthermore, translational regulation of the factors associated with the mRNA 3’- and 5’-end during oocyte and zygote development, will be also studied with the emphasis on finding differences between oocytes and zygotes produced in vitro and in vivo.
Registration No: 19-13491S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Ing. Michal Kubelka, CSc. Institute of Animal Physiology and Genetics CAS, v. v. i.
Other solver: RNDr. Tomáš Mašek, Ph.D. Faculty of Science Charles University
Hybrid sterility and asexuality - two sides of one coin? On the interconnection between asexuality, polyploidy, hybridization and speciation
Basic information: Species are fundamental evolutionary units generally assumed to evolve in a continuum from potentially intermixing populations to independent entities isolated from other species by pre- and postzygotic barriers, such as hybrid sterility or inviability. It was recognized long ago that asexual reproduction often arises as side-effect of hybridization. Our recent research on Cobitis (Teleostei) demonstrated that hybrid asexuality in fact represents an inherent stage of speciation continuum, which prevents the gene flow, but tends to arise earlier during species diversification than sterility or inviability. Asexuality may therefore play a role of primary reproductive barrier during speciation. However, hybrid asexuality appears to arise asymmetrically. While females are fertile and have gametogenetic aberrations alleviating the problems in chromosomal pairings, hybrid males are sterile and lack such an aberration, consequently leading to meiotic arrest due to pairing malformations. We will examine the relation between hybrid sterility and asexuality and their role in speciation.
Registration No: 19-21552S
Project duration: 1. 1. 2019 – 31. 12. 2021, extended to 30. 6. 2022
Principal Investigator: Mgr. Karel Janko, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: FasL/Fas system was identified in apoptotic elimination of cells, however, there is an emerging evidence about non-apoptotic FasL functions. Recently, sclerostin (Sost) expression was found to be decreased in the mandibular bone of FasL deficient mice by the Czech Team thereby suggesting a novel non-apoptotic function of FasL. Consequently, Sost tooth/periodontium phenotype was published by the Austria Team. The overall research aims of this joint project are: (i) To experimentally confirm Sost expression modulation by FasL, (ii) to experimentally challenge possible reciprocal interactions between FasL and Sost, (iii) to decipher the molecular mechanism(s) behind FasL and Sost interactions, (iv) to characterize the dental and periodontal phenotype of FasL knockout mice, (v) to study healing of extraction sockets in FasL knockout mice, (vi) to investigate the role of FasL in regulation of lipopolysaccharide (LPS)-induced periodontitis in mice. The main focus is basic research, which however would provide data interesting for regenerative dentistry and clinical applications.
Registration No: 19-29667L
Project duration: 1. 1. 2019 – 31. 12. 2021
Investigator: Prof. RNDr. Eva Matalová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Mammalian long bones are formed by endochondral ossification which includes a chondrogenic step and formation of the growth plate allowing for bone elongation. To novel factors in chondrogenesis related to long bones have been recently assigned cysteine proteases, particularly those traditionally associated with apoptosis. The latest data showed their presence within the growth plate where their activation patterns did not overlap with apoptotic cells. Additionally, general inhibition of these molecules in limb derived micromass cultures significantly impacted expression of several osteogenic markers. The major goal of this project is to further investigate the novel functions of pro-apoptotic cysteine proteases beyond apoptosis in long bone development. Data from in vivo temporospatial context related to their activation will be followed by results obtained by targeted modulations ex vivo (long bone cultures) and in vitro (micromass cultures) systems to receive an overview of interactions and osteogenic impact of pro-apoptotic cysteine proteases during endochondral ossification.
Registration No: 19-12023S
Project duration: 1. 1. 2019 – 31. 12. 2021
Principal investigator: Prof. RNDr. Eva Matalová, Ph.D. University of Veterinary Sciences Brno
Other solver: Hervé Lesot Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: The extant sturgeons (order Acipenseriformes), ancient evolutionary lineage of non-teleost Actinopterygii, are most remarkable example of evolution via allopolyploidization. During sturgeon evolution, at least three independent whole genome duplication (WGD) events associated with several hybridization events have occurred and sturgeons possess the highest ploidy diversity among all vertebrates. Being still prone for polyploidization, they represent a suitable model for studying the WGD phenomenon. Unreduced gamete formation observed also in sturgeons, has been recently suggested as a mechanism of evolutionary speciation, contrary to general interpretation of it being an evolutionary mishap. Using sturgeons as model species makes the project highly topical for investigation of evolution of all vertebrates, as well as sturgeon conservation. Results should contribute to uncovering the effect of polyploidy for evolutionary success and adaptation of polyploid individuals to unstable environmental conditions when compared to their “non-polyploidized” progenitors or siblings.
Registration No: 18-09323S
Project duration: 1. 1. 2018 – 31. 12. 2021
Principal investigator: prof. Ing. Martin Flajšhans, Dr. rer, agr. Jihočeská univerzita v Českých Budějovicích, Fakulta rybářství a ochrany vod
Other solver: Prof. Ing. Petr Ráb, DrSc. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: Mammalian oocyte is highly differentiated pluripotent cell, which gives foundation to embryo development. Importantly, fully-grown oocyte becomes transcriptionally inactive, and utilizes only transcripts synthesized and stored during earlier development. In addition to protein-coding messenger RNAs, oocytes and embryos produce a plethora of diverse non-coding RNA molecules. Subpopulation of non-coding RNAs belongs to the short and long non-coding RNA class, the functions of which are becoming recently unveiled as being involved in the regulation of mRNA transcription, subcellular localization, turnover, and translation in the somatic cells, however, their implications for oocyte and early embryo biology and pathophysiology is still far from being understood. The goal of this proposal is to shed light on the physiological function of subset of short and long non-coding RNAs in the mammalian oocyte and preimplantation embryo development. We will analyze contribution of selected ncRNAs to the molecular physiology of mammalian oocyte and early embryo.
Registration No: 18-19395S
Project duration: 1. 1. 2018 – 31. 12. 2021
Investigator: Ing. Andrej Šušor, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Basic information: The transplantation of retinal pigment epithelial (RPE) cells derived from human induced pluripotent stem cells (iPSC), is intensively studied as a promising and ethically acceptable therapy for degenerative retinal diseases, such as age-related macular degeneration. We aim this project to assess human iPSC-derived RPE cell grafts supported on our newly developed nanofibrous carrier in a large-eyed animal model, the minipig. The assessment will include long-term safety and efficacy of the graft with a focus on donor cell survival, ability to maintain host retinal architecture, and interaction with the immune system based on non-invasive imaging techniques and end-stage histopathology analyses. Porcine primary RPE cells will serve as a cost-effective reference model of human RPE cells in development of the delivery technique. The combination of clinically perspective cells and a large-eyed animal model will provide valuable data for further advancement in retinal transplantation research.
Registration No: 18-04393S
Project duration: 1. 1. 2018 – 31. 12. 2021
Investigator: MUDr. Taras Ardan, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
Fate decisions in the dental placode: an investigation into the signalling factors that determine cell fate decisions in the early oral cavity
Basic information: Understanding cell fate decisions is an essential goal in developmental biology with important ramifications for tissue regeneration and repair and our understanding of disease. In this proposal we aim to investigate the decisions that determine fate of cells during development of odontogenic placodes. Recently, research from the lab has shown that the dental lamina (tooth forming region) and neighbouring vestibular lamina (lip furrow forming region) in mice are populated from a united Shh expressing domain. This common origin is highlighted by the ability of the vestibular lamina to develop tooth like structure when signalling pathways are manipulated in transgenic mice, and explains the development of odontomas in this region of the jaw in patients. A similar united anlage forms both tooth and dental glands in reptiles. What signalling molecules control the fate of the cells from initial epithelial anlages, and thereby determine whether they form teeth, glands or labial furrows, is unclear and is the aim of this application.
Registration No: 18-04859S
Project duration: 1. 1. 2018 – 31. 12. 2020
Principal investigator: Abigail Tucker Ústav experimentální medicíny AV ČR, v. v. i.
Other solver: doc. RNDr. Marcela Buchtová, Ph.D. Institute of Animal Physiology and Genetics CAS, v. v. i.
