Extracellular ATP Neurotransmission and Nicotine Sex-Specifically Modulate Habenular Neuronal Activity in Adolescence.
The recent increase in the use of nicotine products by teenagers has revealed an urgent need to better understand the impact of nicotine on the adolescent brain. Here, we sought to examine the actions of extracellular ATP as a neurotransmitter and to investigate whether ATP and nicotinic signaling interact during adolescence. With the GRABATP (G-protein-coupled receptor activation-based ATP sensor), we first demonstrated that nicotine induces extracellular ATP release in the medial habenula, a brain region involved in nicotine aversion and withdrawal. Using patch-clamp electrophysiology, we then demonstrated that activation of the ATP receptors P2X or P2Y1 increases the neuronal firing of cholinergic neurons. Surprisingly, contrasting interactive effects were observed with nicotine exposure. For the P2X receptor, activation had no observable effect on acute nicotine-mediated activity, but during abstinence after 10 d of nicotine exposure, coexposure to nicotine and the P2X agonist potentiated neuronal activity in female, but not male, neurons. For P2Y1 signaling, a potentiated effect of the agonist and nicotine was observed with acute exposure, but not following extended nicotine exposure. These data reveal a complex interactive effect between nicotinic and ATP signaling in the adolescent brain and provide mechanistic insights into extracellular ATP signaling with sex-specific alterations of neuronal responses based on prior drug exposure.SIGNIFICANCE STATEMENT In these studies, it was discovered that nicotine induces extracellular ATP release in the medial habenula and subsequent activation of the ATP purinergic receptors increases habenular cholinergic neuronal firing in the adolescent brain. Interestingly, following extended nicotine exposure, nicotine was found to alter the interplay between purinergic and nicotinic signaling in a sex-specific manner. Together, these studies provide a novel understanding for the role of extracellular ATP in mediating habenular activity and reveal how nicotine exposure during adolescence alters these signaling mechanisms, which has important implications given the high incidence of e-cigarette/vape use by youth.
Pubmed ID: 37821229 RIS Download
Research resources used in this publication
- GraphPad Prism (RRID:SCR_002798) (software resource)
- Scientifica (RRID:SCR_021035) (commercial organization)
- Picospritzer III (RRID:SCR_018152) (instrument resource)
- pClamp (RRID:SCR_011323) (software resource)
- MultiClamp 700B Microelectrode Amplifier (RRID:SCR_018455) (instrument resource)
- Sutter P-1000 Next Generation Micropipette Puller (RRID:SCR_021042) (instrument resource)
- SliceScope Pro 2000 (RRID:SCR_018405) (instrument resource)
- C57BL/6J (RRID:IMSR_JAX:000664) (organism)
- GraphPad Prism (RRID:SCR_002798) (software resource)
- Picospritzer III (RRID:SCR_018152) (instrument resource)
- pClamp (RRID:SCR_011323) (software resource)
- MultiClamp 700B Microelectrode Amplifier (RRID:SCR_018455) (instrument resource)
- Sutter P-1000 Next Generation Micropipette Puller (RRID:SCR_021042) (instrument resource)
- SliceScope Pro 2000 (RRID:SCR_018405) (instrument resource)
- C57BL/6J (RRID:IMSR_JAX:000664) (organism)
- Scientifica (RRID:SCR_021035) (commercial organization)
Additional research tools detected in this publication
None foundAntibodies used in this publication
None foundAssociated grants
- Agency: NIDA NIH HHS, United States
Id: DP1 DA039658 - Agency: NIDA NIH HHS, United States
Id: R01 DA051831
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This is a list of tools and resources that we have found mentioned in this publication.
GraphPad Prism (software resource)
RRID:SCR_002798
Statistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.
View all literature mentionsScientifica (commercial organization)
RRID:SCR_021035
Develops and distributes premium scientific equipment to enhance research of neurons, neuronal circuits and animal behaviour.
View all literature mentionsPicospritzer III (instrument resource)
RRID:SCR_018152
Intracellular microinjection dispense system by Parker Hannifin. Supplies repeatable pressure pulses. Designed for rapid and reproducible ejections of picoliter and nanoliter volumes used in conjunction with intracellular or extracellular studies while avoiding inherent desensitization of nerve cells which accompanies Iontophoretic methodology. System comes complete with high speed valves and necessary tubing assemblies.
View all literature mentionspClamp (software resource)
RRID:SCR_011323
Software suite for electrophysiology data acquisition and analysis by Molecular Devices. Used for the control and recording of voltage clamp, current clamp, and patch clamp experiments. The software suite consists of Clampex 11 Software for data acquisition, AxoScope 11 Software for background recording, Clampfit 11 Software for data analysis, and optional Clampfit Advanced Analysis Module for sophisticated and streamlined analysis.
View all literature mentionsMultiClamp 700B Microelectrode Amplifier (instrument resource)
RRID:SCR_018455
Computer controlled amplifier for current clamp and voltage clamp applications from Molecular Devices, LLC. Amplifier used for electrophysiology and electrochemistry.
View all literature mentionsSutter P-1000 Next Generation Micropipette Puller (instrument resource)
RRID:SCR_021042
P-1000 include diagnostic testing of all puller components, built in error detection of air pressure loss or filament burnout, easy access to ramp test, measurement of jaw temperature, and access to previous pull results with heat on times for each cycle of program.Help topics are pre-loaded to assist with on-site troubleshooting and built-in glossary includes text, pictures, and diagrams explaining terms used in micropipette fabrication. Rotary dial is offered as alternative to keypad for numerical entry.
View all literature mentionsSliceScope Pro 2000 (instrument resource)
RRID:SCR_018405
Electrophysiology system for dual or single patch clamp recording with movable microscope. Consists of SliceScope microscope, motorised XY translation stage, fixed stage platform, and two Patchstar motorised micromanipulators. Manufactured by Scientifica LTD.
View all literature mentionsC57BL/6J (organism)
RRID:IMSR_JAX:000664
Mus musculus with name C57BL/6J from IMSR.
View all literature mentionsGraphPad Prism (software resource)
RRID:SCR_002798
Statistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.
View all literature mentionsPicospritzer III (instrument resource)
RRID:SCR_018152
Intracellular microinjection dispense system by Parker Hannifin. Supplies repeatable pressure pulses. Designed for rapid and reproducible ejections of picoliter and nanoliter volumes used in conjunction with intracellular or extracellular studies while avoiding inherent desensitization of nerve cells which accompanies Iontophoretic methodology. System comes complete with high speed valves and necessary tubing assemblies.
View all literature mentionspClamp (software resource)
RRID:SCR_011323
Software suite for electrophysiology data acquisition and analysis by Molecular Devices. Used for the control and recording of voltage clamp, current clamp, and patch clamp experiments. The software suite consists of Clampex 11 Software for data acquisition, AxoScope 11 Software for background recording, Clampfit 11 Software for data analysis, and optional Clampfit Advanced Analysis Module for sophisticated and streamlined analysis.
View all literature mentionsMultiClamp 700B Microelectrode Amplifier (instrument resource)
RRID:SCR_018455
Computer controlled amplifier for current clamp and voltage clamp applications from Molecular Devices, LLC. Amplifier used for electrophysiology and electrochemistry.
View all literature mentionsSutter P-1000 Next Generation Micropipette Puller (instrument resource)
RRID:SCR_021042
P-1000 include diagnostic testing of all puller components, built in error detection of air pressure loss or filament burnout, easy access to ramp test, measurement of jaw temperature, and access to previous pull results with heat on times for each cycle of program.Help topics are pre-loaded to assist with on-site troubleshooting and built-in glossary includes text, pictures, and diagrams explaining terms used in micropipette fabrication. Rotary dial is offered as alternative to keypad for numerical entry.
View all literature mentionsSliceScope Pro 2000 (instrument resource)
RRID:SCR_018405
Electrophysiology system for dual or single patch clamp recording with movable microscope. Consists of SliceScope microscope, motorised XY translation stage, fixed stage platform, and two Patchstar motorised micromanipulators. Manufactured by Scientifica LTD.
View all literature mentionsC57BL/6J (organism)
RRID:IMSR_JAX:000664
Mus musculus with name C57BL/6J from IMSR.
View all literature mentionsScientifica (commercial organization)
RRID:SCR_021035
Develops and distributes premium scientific equipment to enhance research of neurons, neuronal circuits and animal behaviour.
View all literature mentions
