temporal regulation of gabaa receptor subunit · pdf filetemporal regulation of gabaa receptor...

58
Accepted manuscripts are peer-reviewed but have not been through the copyediting, formatting, or proofreading process. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. Copyright © 2017 the authors This Accepted Manuscript has not been copyedited and formatted. The final version may differ from this version. Research Article: New Research | Integrative Systems Temporal Regulation of GABAA Receptor Subunit Expression: Role in Synaptic and Extrasynaptic Communication in the Suprachiasmatic Nucleus Functional GABAA receptor rhythms in the SCN James C. Walton, John K. McNeill, Khallyl A. Oliver and H. Elliott Albers Neuroscience Institute and Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303, USA DOI: 10.1523/ENEURO.0352-16.2017 Received: 28 November 2016 Revised: 11 April 2017 Accepted: 12 April 2017 Published: 21 April 2017 Author Contributions: JCW and HEA designed the research, analyzed the data, and wrote the manuscript. JCW, JKM, and KAO performed the research. Funding: HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS) 100000065 F32NS092545 Funding: HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS) 100000065 R01NS078220 The authors declare no competing financial interests. National Institutes of Health grant R01NS078220 to HEA and F32NS092545 to JCW. Correspondence should be addressed to James C. Walton, Neuroscience Institute and Center for Behavioral Neuroscience, Georgia State University, PO Box 5030, Atlanta, GA 30303, USA, E-mail: [email protected] Cite as: eNeuro 2017; 10.1523/ENEURO.0352-16.2017 Alerts: Sign up at eneuro.org/alerts to receive customized email alerts when the fully formatted version of this article is published.

Upload: buinga

Post on 19-Feb-2018

228 views

Category:

Documents


6 download

TRANSCRIPT

Page 1: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Accepted manuscripts are peer-reviewed but have not been through the copyediting, formatting, or proofreadingprocess.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International(http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in anymedium provided that the original work is properly attributed.

Copyright © 2017 the authors

This Accepted Manuscript has not been copyedited and formatted. The final version may differ from this version.

Research Article: New Research | Integrative Systems

Temporal Regulation of GABAA Receptor Subunit Expression: Role in Synapticand Extrasynaptic Communication in the Suprachiasmatic Nucleus

Functional GABAA receptor rhythms in the SCN

James C. Walton, John K. McNeill, Khallyl A. Oliver and H. Elliott Albers

Neuroscience Institute and Center for Behavioral Neuroscience, Georgia State University, Atlanta, GA 30303,USA

DOI: 10.1523/ENEURO.0352-16.2017

Received: 28 November 2016

Revised: 11 April 2017

Accepted: 12 April 2017

Published: 21 April 2017

Author Contributions: JCW and HEA designed the research, analyzed the data, and wrote the manuscript.JCW, JKM, and KAO performed the research.

Funding: HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS)100000065F32NS092545

Funding: HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS)100000065R01NS078220

The authors declare no competing financial interests.

National Institutes of Health grant R01NS078220 to HEA and F32NS092545 to JCW.

Correspondence should be addressed to James C. Walton, Neuroscience Institute and Center for BehavioralNeuroscience, Georgia State University, PO Box 5030, Atlanta, GA 30303, USA, E-mail: [email protected]

Cite as: eNeuro 2017; 10.1523/ENEURO.0352-16.2017

Alerts: Sign up at eneuro.org/alerts to receive customized email alerts when the fully formatted version of thisarticle is published.

Page 2: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 3: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 4: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 5: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 6: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

e.g e.g

Page 7: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Animals and housing.

Mesocricetus auratus

ad

libitum

Page 8: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Experiment 1: Effects of GABAA receptor subtype-specific agonists on phase

resetting.

Page 9: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

in vivo

Page 10: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Experiment 2: GABAA receptor subunit gene expression in the SCN.

Page 11: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Experiment 3: GABAA receptor subunit protein expression in the SCN.

Page 12: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 13: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 14: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

Statistics.

t

Page 15: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

t

t

p

Experiment 1: Phase shifting effects of GABAA agonists.

F p

F p

F p

p

p

Experiment 2: GABAA receptor subunit gene expression in the SCN.

Page 16: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

t

t p

t

p

t p

t p

Experiment 3: GABAA receptor subunit protein immunoreactivity in the SCN.

Page 17: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

t

Page 18: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

p

p

i.e.

Page 19: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

F p

F p

F p

Page 20: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

i.e

t

t

Page 21: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

lowest

Page 22: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

ratio

Page 23: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

synaptic

phasic extrasynaptic tonic

vs

sustained

Page 24: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

clock

clock

Page 25: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 26: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 27: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 28: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 29: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 30: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 31: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 32: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 33: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 34: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 35: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 36: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 37: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 38: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

p

Page 39: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p p

Page 40: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

p

p

Page 41: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p

Page 42: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

F p

Treatment vs Treatment p

Page 43: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

p <

Page 44: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

F t p

2

: 2

2

: 2

2

: 2

2

: 2 p <

Page 45: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

F t p

2

: 2

2

: 2

2

: 2

2

: 2 p <

Page 46: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

F p F p F p

WHOLE

DORSAL

VENTRAL

ROSTRAL

CENTRAL ANTERIOR

CENTRAL POSTERIOR

CAUDAL

DORSAL ROSTRAL

DORSAL CENTRAL ANTERIOR

Page 47: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

DORSAL CENTRAL POSTERIOR

DORSAL CAUDAL

VENTRAL ROSTRAL

VENTRAL CENTRAL ANTERIOR

VENTRAL CENTRAL POSTERIOR

VENTRAL CAUDAL

Page 48: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

t

t p

2

2

2

p <

Page 49: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

t p

2

Page 50: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV

2

2

2

2

2

p <

Page 51: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 52: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 53: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 54: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 55: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 56: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 57: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV
Page 58: Temporal Regulation of GABAA Receptor Subunit · PDF fileTemporal Regulation of GABAA Receptor Subunit Expression: ... tvx %UDLQVZHUHVHFWLRQHGDW PRQDFU\RVWDW DQGWKUHHVHWVRI VHULDOFRURQDOVHFWLRQV