Liposome-entrapped GABA modulates the expression of nNOS in NG108-15 cells

Gisele C. Vaz, Neeru M. Sharma, Hong Zheng, Matthew C Zimmerman, Robson S. Santos, Frederic Frezard, Marco A P Fontes, Kaushik P Patel

Research output: Contribution to journalArticle

4 Citations (Scopus)

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 24 h showed an increase in expression of GABAAR (54 and 50% with 10 and 20 ng doses, respectively) and nNOS (138, 157 and 165% with 20, 50 and 100 ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50 ng of GL showed an increase in GABAAR (23%) after 1 h followed by an increase in nNOS (55, 46 and 55%) at 8, 12 and 24 h 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 100 ng 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.

Original languageEnglish (US)
Pages (from-to)55-63
Number of pages9
JournalJournal of Neuroscience Methods
Volume273
DOIs
StatePublished - Nov 1 2016

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Nitric Oxide Synthase Type I
Liposomes
gamma-Aminobutyric Acid
GABA-A Receptors
Neurotransmitter Agents
Aminobutyrates
Fluorescent Antibody Technique
Nitric Oxide
Proteins
Central Nervous System
Lipids
Cell Line

Keywords

  • GABA
  • Liposome
  • NG108-15
  • PIN
  • nNOS

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Liposome-entrapped GABA modulates the expression of nNOS in NG108-15 cells. / Vaz, Gisele C.; Sharma, Neeru M.; Zheng, Hong; Zimmerman, Matthew C; Santos, Robson S.; Frezard, Frederic; Fontes, Marco A P; Patel, Kaushik P.

In: Journal of Neuroscience Methods, Vol. 273, 01.11.2016, p. 55-63.

Research output: Contribution to journalArticle

Vaz, Gisele C. ; Sharma, Neeru M. ; Zheng, Hong ; Zimmerman, Matthew C ; Santos, Robson S. ; Frezard, Frederic ; Fontes, Marco A P ; Patel, Kaushik P. / Liposome-entrapped GABA modulates the expression of nNOS in NG108-15 cells. In: Journal of Neuroscience Methods. 2016 ; Vol. 273. pp. 55-63.
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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 24 h showed an increase in expression of GABAAR (54 and 50{\%} with 10 and 20 ng doses, respectively) and nNOS (138, 157 and 165{\%} with 20, 50 and 100 ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50 ng of GL showed an increase in GABAAR (23{\%}) after 1 h followed by an increase in nNOS (55, 46 and 55{\%}) at 8, 12 and 24 h 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 100 ng 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.",
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AU - Sharma, Neeru M.

AU - Zheng, Hong

AU - Zimmerman, Matthew C

AU - Santos, Robson S.

AU - Frezard, Frederic

AU - Fontes, Marco A P

AU - Patel, Kaushik P

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N2 - 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 24 h showed an increase in expression of GABAAR (54 and 50% with 10 and 20 ng doses, respectively) and nNOS (138, 157 and 165% with 20, 50 and 100 ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50 ng of GL showed an increase in GABAAR (23%) after 1 h followed by an increase in nNOS (55, 46 and 55%) at 8, 12 and 24 h 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 100 ng 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.

AB - 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 24 h showed an increase in expression of GABAAR (54 and 50% with 10 and 20 ng doses, respectively) and nNOS (138, 157 and 165% with 20, 50 and 100 ng doses, respectively) compared with cells treated with empty liposomes (EL). Additionally, cells treated with 50 ng of GL showed an increase in GABAAR (23%) after 1 h followed by an increase in nNOS (55, 46 and 55%) at 8, 12 and 24 h 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 100 ng 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.

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