Neuron-astrocyte interactions

TalTech priority area
Research classification (Frascati)
Head of the research group
Doctoral students
cell types of the nervous system
cell type-specific RNA and protein profiling
neurotrophin BDNF in non-neuronal cells
The central nervous system tissues are made ofa number of different cell types, among whichastrocytes are one of the most abundant type.In the CNS tissue cells are highly intermixed,posing a challenge when trying to analyze theirtranscriptomes and proteomes separately. Owing to the difficulties separating these cells,bulk tissue analysis has been used previouslyto profile mRNA and protein in tissue, givingaveraged readouts across the tissue. In the pastdecade, cell type specific RNA analysis has seenenormous progress with the advent of singlecell RNA sequencing and genetic tools for celltype specific RNA isolation (TRAP, Ribotag).However, cell type specific proteome analysis is lagging behind and widely used, straightforwardmethods are not available.Our research aim is to develop a cell type-specific proteome analysis method that is based onpuromycin labeling, and to apply the method tostudying neuron-astrocyte interactions in an invitro co-culture system. In addition, we shall usethe Ribotag method for cell type-specific mRNAanalysis.A key strategy we are planning to use for studying intercellular communication is activation ofeither neurons of astrocytes by triggering intracellular Ca2+ release by using the DREADDchemogenetic system – followed by proteomicand transcriptomic analysis of the other celltype in culture.Our additional research interest is regulationof neurotrophin BDNF in astrocytes and cardiomyocytes.Key competences: cell cultures of neurons, astrocytes and cardiomyocytes; cell type-specific RNAand protein analysis; adeno-associated virus(AAV) vector production and use
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.
Period of activity of the research group
Teadusgrupiga seotud publikatsioonid