Projects / Programmes
Dynamic measurements of metabolites and second messengers in single cultured astrocytes
| Code |
Science |
Field |
Subfield |
| 3.03.00 |
Medical sciences |
Neurobiology |
|
| Code |
Science |
Field |
| B000 |
Biomedical sciences |
|
| Code |
Science |
Field |
| 3.01 |
Medical and Health Sciences |
Basic medicine |
Astrocytes, Metabolism, L-Lactate, cAMP, D-Glucose, Intelligent Deficiency, Neurodegeneration
Researchers (17)
Organisations (2)
Abstract
PROPOSAL ABSTRACT-SUMMARY
From the initial consideration of being the “brain glue”, astrocytes, the most abundant glial cells in the brain, are today regarded as multifunctional cells implicated in a myriad of processes including the role of integrating and tuning inter-neuronal signalling through the uptake and release of several chemical messengers – gliotransmitters. Some of the gliotransmitters are stored in vesicles and it is becoming clear that the properties of Ca2+-dependent fusion of the vesicle and the plasma membranes (i.e. exocytotic fusion) differs in astrocytes and in neurons mainly in the kinetics. It is orders of magnitude slower in astrocytes in comparison with neurons. The relatively slow kinetics of vesicle based signalling in astrocytes likely involves vesicle dynamics and mechanisms governing the merger of the vesicle membrane with the plasma membrane. Moreover, it may also depend on various metabolic pathways, which are determining the availability of precursors for gliotransmitters such as ATP and glutamate, and their packaging into vesicles via specific vesicle transporters. The aim of this proposal is to study the regulation of metabolites in single astrocytes, such as glucose, lactate and the second messenger cAMP, which regulates the degradation of glycogen. It has been hypothesized that when lactate is produced in astrocytes, this exits into the extracellular space via the monocarboxylic acid transporters (MCTs) and binds to extracellular plasma membrane lactate receptors. Autocrine activation of lactate receptors on astrocytes may regulate the metabolism of astrocytes via a feed-back mechanism. To directly verify these questions we will use FRET-based nanosensors expressed in astrocytes to detect in real-time the cytosolic concentration of cAMP, D-glucose and L-lactate in single living astrocytes, prepared in primary culture, by the use of advanced quantitative fluorescence microscopy. We will also address whether the dynamics of cytosolic concentration of cAMP, D-glucose and L-lactate is altered in astrocytes from a mouse model of Intelligent Deficiency. All these novel approaches represent the methods of choice to address these questions. Therefore, we strongly believe that the results of the proposed research are not only novel, but will provide important new insights into the understanding of astrocyte role in physiological and pathological conditions, such as mental retardation and neurodegeneration.
Significance for science
In the focus of our research were astrocytes, the most abundant glial cells in the mammalian brain which have the crucial support role for the functioning of neurons. The role of astrocytes was studied from the perspective of the regulation of aerobic glycolsis and vesicle traffic by membrane bound vesicles. Studies in recent years underligned a connection between the impaired vesicle traffic in several diseases, including neurodegeneration of the central nervous system (CNS) and cognitive disabilities. While until recently the studies were fucused into neurons, the key players in the CNS, more and more evidence highlights the important role of astrocytes, neuronal supportive cells. Our studies have revealed new mechanisms of regulation of aerobic glycolysis in astrocytes.
Significance for the country
Fundamental scientific research in the field of cell phisology is essential to elucidate how cells operate at tissue and organ levels, in normal and pathological states This is an important platform for discovery and improvements of diagnostic procedures and the development of novel, more specific targets and drugs, all adding to the capacity building of the country at international level. Moreover, international visibility is accomplished through publishing the research results in peer reviewed journals and by the active attendance at international meetings, all recognizing our research group and the R. of Slovenia. Sciences also positively influence the educational sector and the recruitment of younger reseachers and students. This research project increased the visibility of our research group members in international scientific circles and contributed to the streamlining of added value for the next period of facilitated research activities. In all increasing also the international visibility of the country.
Most important scientific results
Annual report
2014,
2015,
final report
Most important socioeconomically and culturally relevant results
Annual report
2014,
2015,
final report
