title: Characterization of Amino Acid Profile and Enzymatic Activity in Adult Rat Astrocyte Cultures.
abstract: Astrocytes are multitasking players in brain complexity, possessing several receptors and mechanisms to detect, participate and modulate neuronal communication. The functionality of astrocytes has been mainly unraveled through the study of primary astrocyte cultures, and recently our research group characterized a model of astrocyte cultures derived from adult Wistar rats. We, herein, aim to characterize other basal functions of these cells to explore the potential of this model for studying the adult brain. To characterize the astrocytic phenotype, we determined the presence of GFAP, GLAST and GLT 1 proteins in cells by immunofluorescence. Next, we determined the concentrations of thirteen amino acids, ATP, ADP, adenosine and calcium in astrocyte cultures, as well as the activities of Na(+)/K(+)-ATPase and acetylcholine esterase. Furthermore, we assessed the presence of the GABA transporter 1 (GAT 1) and cannabinoid receptor 1 (CB 1) in the astrocytes. Cells demonstrated the presence of glutamine, consistent with their role in the glutamate-glutamine cycle, as well as glutamate and D-serine, amino acids classically known to act as gliotransmitters. ATP was produced and released by the cells and ADP was consumed. Calcium levels were in agreement with those reported in the literature, as were the enzymatic activities measured. The presence of GAT 1 was detected, but the presence of CB 1 was not, suggesting a decreased neuroprotective capacity in adult astrocytes under in vitro conditions. Taken together, our results show cellular functionality regarding the astrocytic role in gliotransmission and neurotransmitter management since they are able to produce and release gliotransmitters and to modulate the cholinergic and GABAergic systems.
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil. debsrs@gmail.com.
authors.forename: Débora Guerini
authors.initials: DG
authors.lastname: Souza
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Bruna
authors.initials: B
authors.lastname: Bellaver
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Gisele
authors.initials: G
authors.lastname: Hansel
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Bernardo Assein
authors.initials: BA
authors.lastname: Arús
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Bellaver
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Aline
authors.initials: A
authors.lastname: Longoni
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Janaina
authors.initials: J
authors.lastname: Kolling
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Angela T S
authors.initials: AT
authors.lastname: Wyse
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: Diogo Onofre
authors.initials: DO
authors.lastname: Souza
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600, Anexo, PO Box: 90035-003, Porto Alegre, RS, Brazil.
authors.forename: André
authors.initials: A
authors.lastname: Quincozes-Santos
mesh_terms: D000110:Acetylcholinesterase / Q000032:analysis* / Q000378:metabolism; D000367:Age Factors; D000596:Amino Acids / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D050485:GABA Plasma Membrane Transport Proteins / Q000032:analysis / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D000254:Sodium-Potassium-Exchanging ATPase / Q000032:analysis* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D050485:GABA Plasma Membrane Transport Proteins; D000110:Acetylcholinesterase; D000254:Sodium-Potassium-Exchanging ATPase
keywords: AChE; Adult astrocytes; Amino acids; Gliotransmitters; Na+/K+-ATPase