Gisele C Vaz1, Neeru M Sharma2, Hong Zheng2, Matthew C Zimmerman2, Robson S Santos1, Frederic Frezard1, Marco A P Fontes1, Kaushik P Patel3. 1. Department of Physiology & Biophysics, Federal University of Minas Gerais, Belo Horizonte, Brazil. 2. Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE 68198-5850, United States. 3. Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, NE 68198-5850, United States. Electronic address: kpatel@unmc.edu.
Abstract
BACKGROUND: Liposomes are concentric lipid vesicles that allow a sustained release of entrapped substances. GABA (γ-aminobutyric acid) is the most prevalent inhibitory neurotransmitter in the central nervous system. NEW METHOD: Using GABA-containing liposomes (GL) prepared by the freeze-thawing method, we determined the effect of sustained release of GABA on expression of neuronal nitric oxide synthase (nNOS) and GABAA receptor (GABAAR) in an in vitro neuronal model. RESULTS: Neuronal cell line NG108-15 treated with different doses of GL during 24h showed an increase in expression of GABAAR (54 and 50% with 10 and 20ng doses, respectively) and nNOS (138, 157 and 165% with 20, 50 and 100ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50ng of GL showed an increase in GABAAR (23%) after 1h followed by an increase in nNOS (55, 46 and 55%) at 8, 12 and 24h time points, respectively. Immunofluorescence experiments confirmed an increase in nNOS (134%) and basal intracellular levels of nitric oxide (84%) after GL treatment. Further, treatment of cells with GL showed a decrease in expression of a protein inhibitor of nNOS (PIN) (26, 66 and 57% with 20, 50 and 100ng doses respectively) compared with control. COMPARISON WITH EXISTING METHODS: This is first demonstration for the development of GL that allows sustained slow release of this neurotransmitter. CONCLUSION: These results suggest that a slow release of GABA can change the expression of nNOS possibly via alteration in PIN levels in neuronal cells.
BACKGROUND: Liposomes are concentric lipid vesicles that allow a sustained release of entrapped substances. GABA (γ-aminobutyric acid) is the most prevalent inhibitory neurotransmitter in the central nervous system. NEW METHOD: Using GABA-containing liposomes (GL) prepared by the freeze-thawing method, we determined the effect of sustained release of GABA on expression of neuronal nitric oxide synthase (nNOS) and GABAA receptor (GABAAR) in an in vitro neuronal model. RESULTS: Neuronal cell line NG108-15 treated with different doses of GL during 24h showed an increase in expression of GABAAR (54 and 50% with 10 and 20ng doses, respectively) and nNOS (138, 157 and 165% with 20, 50 and 100ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50ng of GL showed an increase in GABAAR (23%) after 1h followed by an increase in nNOS (55, 46 and 55%) at 8, 12 and 24h time points, respectively. Immunofluorescence experiments confirmed an increase in nNOS (134%) and basal intracellular levels of nitric oxide (84%) after GL treatment. Further, treatment of cells with GL showed a decrease in expression of a protein inhibitor of nNOS (PIN) (26, 66 and 57% with 20, 50 and 100ng doses respectively) compared with control. COMPARISON WITH EXISTING METHODS: This is first demonstration for the development of GL that allows sustained slow release of this neurotransmitter. CONCLUSION: These results suggest that a slow release of GABA can change the expression of nNOS possibly via alteration in PIN levels in neuronal cells.
Authors: G C Vaz; A P C O Bahia; F C de Figueiredo Müller-Ribeiro; C H Xavier; K P Patel; R A S Santos; F A Moreira; F Frézard; M A P Fontes Journal: Neuroscience Date: 2014-11-13 Impact factor: 3.590
Authors: Giuseppe Corace; Cristina Angeloni; Marco Malaguti; Silvana Hrelia; Paul C Stein; Martin Brandl; Roberto Gotti; Barbara Luppi Journal: J Liposome Res Date: 2014-05-07 Impact factor: 3.648
Authors: Marco Antônio Peliky Fontes; Gisele Cristiane Vaz; Thais Zielke Dias Cardoso; Mariana Flávia de Oliveira; Maria José Campagnole-Santos; Robson Augusto Souza Dos Santos; Neeru M Sharma; Kaushik P Patel; Frédéric Frézard Journal: Nanomedicine Date: 2017-12-24 Impact factor: 5.307
Authors: Timofey D Lebedev; Elmira R Vagapova; Vladimir I Popenko; Olga G Leonova; Pavel V Spirin; Vladimir S Prassolov Journal: Front Oncol Date: 2019-10-18 Impact factor: 6.244