Glial cell biology
Nimetus
Neuron-astrocyte interactions (until 2023)
Head of the research group
Related department
TalTech priority area
Overview
The central nervous system tissues are made of a number of different cell types, among which astrocytes are one of the most abundant type. In the CNS tissue cells are highly intermixed, posing a challenge when trying to analyze their transcriptomes and proteomes separately. Owing to the difficulties separating these cells, bulk tissue analysis has been used previously to profile mRNA and protein in tissue, giving averaged readouts across the tissue. In the past decade, cell type specific RNA analysis has seen enormous progress with the advent of single cell RNA sequencing and genetic tools for cell type specific RNA isolation (TRAP, Ribotag). However, cell type specific proteome analysis is lagging behind and widely used, straightforward methods are not available. Our research aim is to develop a cell type-specific proteome analysis method that is based on puromycin labeling, and to apply the method to studying neuron-astrocyte interactions in an in vitro co-culture system. In addition, we shall use the Ribotag method for cell type-specific mRNA analysis. A key strategy we are planning to use for studying intercellular communication is activation of either neurons of astrocytes by triggering intracellular Ca2+ release by using the DREADD chemogenetic system – followed by proteomic and transcriptomic analysis of the other cell type in culture. Our additional research interest is regulation of neurotrophin BDNF in astrocytes and cardiomyocytes. Key competences: cell cultures of neurons, astrocytes and cardiomyocytes; cell type-specific RNA and protein analysis; adeno-associated virus(AAV) vector production and use
Research group member
Doctoral students
Research classification (Frascati)
Keyword
cell types of the nervous system
cell type-specific RNA and protein profiling
neurotrophin BDNF in non-neuronal cells
Important results
2021 RESULTS: We established a prenatal rat brain cortexneuron-astrocyte co-culture system anda method for cell-specific effector expression, using selective promoters and AAVtransduction. In comparison of two cell-specific puromycin based proteome tagging methods, weselected a strategy based on enzymaticinactivation of puromycin and its derivatives. We established a prenatal rat cardiomyocyte culture protocol and obtainedpreliminary results in BDNF regulationby catecholamines in these cells.
- Doron-Mandel, E., Koppel, I., Abraham, O., Rishal, I., et al. The glycine arginine-rich domain of the RNA-binding protein nucleolin regulates its subcellular localization // The EMBO Journal (2021) vol. 40, 20, art. e107158.
https://doi.org/10.15252/embj.2020107158 - Koppel, I., Fainzilber, M. Omics approaches for subcellular translation studies // Molecular omics (2018) vol. 14, 6, p. 380–388 : ill.
http://dx.doi.org/10.1039/c8mo00172c - Urb, M., Anier, K., Matsalu, T., Koppel, I., Timmusk, T. et al. Glucocorticoid receptor stimulation resulting from early life stress affects expression of DNA methyltransferases in rat prefrontal cortex // Journal of molecular neuroscience (2019) vol. 68, p. 99–110 : ill.
https://doi.org/10.1007/s12031-019-01286-z - Rozenbaum, M., Rajman, M., Rishal, I., Koppel, I. et al. Translatome regulation in neuronal injury and axon regrowth // eNeuro (2018) vol. 5, 2, art. e0276-17.2018, 15 p. : ill.
http://dx.doi.org/10.1523/ENEURO.0276-17.2018 - Koppel, I., Jaanson, K., Klasche, A., Tuvikene, J., Tiirik, T., Pärn, A., Timmusk, T. Dopamine cross‐reacts with adrenoreceptors in cortical astrocytes to induce BDNF expression, CREB signaling and morphological transformation // GLIA (2018) vol. 66, 1, p. 206-216 : ill.
https://doi.org/10.1002/glia.23238 - Terenzio, M., Koley, S., Samra, N., Koppel, I. et al. Locally translated mTOR controls axonal local translation in nerve injury // Science (2018) vol. 359, 6382, p. 1416-1421 : ill.
http://dx.doi.org/10.1126/science.aan1053 - Lekk, I., Cabrera-Cabrera, F., Turconi, G., Tuvikene, J., Esvald, E.-E., Rähni, A., Casserly, L., Garton, D. R., Andressoo, J.-O., Timmusk, T., Koppel, I. Untranslated regions of brain-derived neurotrophic factor (Bdnf) mRNA control its translatability and subcellular localization // The journal of biological chemistry (2023) vol. 299, 2, art. 102897.
https://doi.org/10.1016/j.jbc.2023.102897 - Esvald, E-E., Tuvikene, J., Kiir, C. S., Avarlaid, A., Tamberg, L., Sirp, A., Shubina, A., Cabrera-Cabrera, F., Pihlak, A., Koppel, I., Palm, K., Timmusk, T. Revisiting the expression of BDNF and its receptors in mammalian development // Frontiers in Molecular Neuroscience (2023) vol. 16, art. 1182499.
https://doi.org/10.3389/fnmol.2023.1182499 - Cabrera-Cabrera, F., Tull, H., Capuana, R., Kasvandik, S., Timmusk, T., Koppel, I. Cell type-specific labelling of newly synthesized proteins by puromycin inactivation // Journal of biological chemistry (2023) vol. 299, 9, art. 105129, 12 p. : ill.
https://doi.org/10.1016/j.jbc.2023.105129 - Avarlaid, A., Esvald, E‐E., Koppel, I., Parkman, A., Zhuravskaya, A., Makeyev, E. V., Tuvikene, J., Timmusk, T. An 840 kb distant upstream enhancer is a crucial regulator of catecholamine-dependent expression of the BDNF gene in astrocytes
// Glia (2023) vol. 72(1), p. 90-110 : ill.https://doi.org/10.1002/glia.24463