The aim of the course is to teach about inner cell organization and cell response on external and internal cues and its communication with the environment.
1. Introduction: basic structure of eukaryotic cell, endosymbiotic theory and theory of information
Types of molecular transport in the cell. Endoplasmic reticulum (ER): targeting proteins to and across the ER, protein folding and quality control in the ER, protein N glycosylation, functions of smooth ER, lipid synthesis, vesicular traffic between ER and Golgi
2.Structure and function of Golgi apparatus. protein glycosylation, lysosome structure, protein targeting to lysosomes, lysosomal storage diseases, mechanisms of retrograde transport, vesicle structure, Rab and SNARE molecules, endocytic and exocytic pathways, vesicle trafficking in polar cells, transcellular transport.
3.Posttranslational transport. Targeting of proteins to mitochondria and chloroplasts, structure and formation of peroxisomes, targeting of peroxisomal proteins, peroxisome functions.
Specific protein synthesis and vesicular traffic in hematopoietic cells and in the defence against microorganisms.
4.Structure and function of the nucleus. Structure of the nuclear envelope and lamina, link between cytosol cytoskeleton and chromatin through transmembrane proteins of nuclear membrane and nuclear matrix, mechanisms of nuclear envelope breakdown and reconstitution during mitosis, laminopathies, transport into and out of the nucleus, regulation of transcription through localization in the nucleus, nuclear bodies.
5.Cytoskeleton. Basic principles of organization and common feature of three types of cytoskeleton. Actin cytoskeleton: basic structure, the role of regulatory proteins, molecular motors, regulation of skeletal muscle contraction, cell migration, Rho family proteins, cell polarization and intercellular connections.
6.Microtubules. Basic structure and organization of microtubules, regulatory proteins, molecular motors kinesins and dyneins and their role in the cell: organelle trafficking and localization, structure of the cilium, centrosome, centrioles, ciliopathies
Intermediate filaments: types and specificities, regulation of lamin polymerization, desmosome structure, neurofilaments.
7.Cell cycle regulation: discovery of the cell cycle mechanism, structure and regulation of maturation promotion factor, cycline B/cdc2, downstream processes leading to entrance in mitosis, restriction point and regulation of entrance in S phase, mechanisms of control of DNA replication and proper chromosome segregation in mitosis, regulation on control points in G1 and G2 phase.
Mechanics of mitosis. Processes in each phase of mitosis and their regulation, asymmetric division.
8.Extracellular matrix: basic structure of proteoglycans and fibrilar proteins, structure of proteoglycans, collagen, laminin, fibronectin; structure and function of basal lamina; signalling pathways by which extracellular matrix components regulate the cellular processes and by which the cell shape the extracellular matrix.
Cell-cell and cell-matrix adhesion. Structure and function of tight junctions, gap junctions, adhesion junctions
9. Signalling in the cell: basic pathways of signalling in the organism, discovery of the growth factors, basic signaling pathways: MAP kinase pathway, survival pathways, Calcium as a messenger, pathways connected with cytoskeleton.
10. Tumour suppressors: basic functions, history of their discovery, pRb, molecules involved in DNA damage repair as tumour suppressors and their connection with inherited diseases, molecules in developmental pathways as tumour suppressors. Tumour suppressors in the regulation of cell death.
Mechanism of p53 function. PoliADPribosylation
11. Regulation of gene expression:. regulation of transcription, transcription factors, chromatin remodelling by molecular remodelators and noncoding RNA, principle of combinatorics and regulatory circles
12. Cell differentiation. Characteristics and types of stem cells, regulation of stem cell maintenance, mechanisms of establishment of induced pluripotent cells and other ways of cellular reprogramming, somatic stem cell niches in the organism, principles of differentiation and morphogens, asymmetric cell division and cellular polarization
13. Aging. Structure and function of telomeres in senescence and carcinogenesis. Senescence on the level of the cell and organism. Aging as an evolution strategy.
Exercises are organised in the way that allow individual practical work. They are based on the cell culture techniques. Individual exercises are complete experiments which comprise cell cultivation, their treatment and analysis of collected data. At the end, students are obliged to write a final report which is analysed together.
1. Cell cycle analysis by flow cytometry and cell synchronization in the culture
2. cellular DNA synthesis follow up by incorporation of labelled DNA precursor molecules
3. apoptosis follow up by staining the cells with fluorescent dyes and their analysis under fluorescent microscope
4. detection of senescent cells in culture
5. cell fusion
6. fluorescent microscopy of cell organelles labelled by molecules fused with green fluorescent protein
7. follow up of autophagy by acridine orange staining
8. analysis of cell tumorigenicity by growth in soft agar
Each student will have a seminar made according to scientific article on the topic connected with the lectures which is broadening certain aspects of given material.
to distinguish between different mechanisms of regulation of intracellular trafficking in the eukaryotic cell
to explain the mechanisms of regulation of cellular cytoskeleton
to connect the structure and the function of inner cell organization with the mechanism of regulation in dependence on internal and external cues
to explain the pathways of cell cycle regulation and mitosis
to explain the mechanisms of cellular signalling and reaction on extracellular cues in cell communication with its environment
to analyze the pathways of cellular processes during differentiation, senescence and cell death
to apply the techniques and theoretical knowledge in experiment setup in cell culture
the analysis of the data obtained by experimental work on exercises in cell culture
- Alberts, B., Bray, D., Lewis, J. Raff, M., Roberts, K., Watson, J.D. (2002):.Molecular biology of the Cell, Garland Publ. Inc., New York, London, IV. izd.
Cooper, G. M. Hausman R. E.(2002): Stanica - molekularni pristup. Medicinska naklada Zagreb.
- Lodish, H., Berk, A., Zipursky, S. L., Matsudaira, P., Baltimore, Darnell, J E. (2000): Molecular Cell Biology. 4th iz. W. H. Freeman & Co, New York.
Ambriović Ristov A (ur.) (2007): Metode u molekularnoj biologiji. Institut Ruđer Bošković, Zagreb.
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