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Articles, Systems/Circuits

Subsecond Regulation of Synaptically Released Dopamine by COMT in the Olfactory Bulb

Renee Cockerham, Shaolin Liu, Roger Cachope, Emi Kiyokage, Joseph F. Cheer, Michael T. Shipley and Adam C. Puche
Journal of Neuroscience 20 July 2016, 36 (29) 7779-7785; https://doi.org/10.1523/JNEUROSCI.0658-16.2016
Renee Cockerham
1University of Maryland School of Medicine, Baltimore, Maryland 21201,
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Shaolin Liu
1University of Maryland School of Medicine, Baltimore, Maryland 21201,
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Roger Cachope
2CHDI Foundation/CHDI Management, New York, New York 10001, and
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Emi Kiyokage
3Kawasaki Medical School, Matsushima, Kurashiki-City, Okayama
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Joseph F. Cheer
1University of Maryland School of Medicine, Baltimore, Maryland 21201,
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Michael T. Shipley
1University of Maryland School of Medicine, Baltimore, Maryland 21201,
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Adam C. Puche
1University of Maryland School of Medicine, Baltimore, Maryland 21201,
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    Figure 1.

    COMT expression. A, SDS-PAGE blot of DAT in the MOB (100 μg in lane) and STR (10 μg in lane) showing the expected molecular weight for DAT. B, Micro-isolated layers of the OB stained for DAT showing the highest DAT expression in the GL, with little expression in the EPL and none in the deeper MCL and GCL. C, Expression of COMT in wild-type (+/+) and null mutant (−/−). D, Micro-isolated layers of the OB and whole STR stained for COMT showing the highest intensity in the glomerular layer, but also expression in all deeper layers. E, Quantification of the relative abundance of COMT in layers of the olfactory bulb and striatum [relative optical density (OD) U/μg total protein]. F, Immunohistochemistry for COMT in the OB. As expected from Western blot data, expression is present throughout the OB with the exception of the ONL, which is devoid of significant staining. Scale bar, 80 μm.

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    Figure 2.

    Quantification of TH, glutamic acid decarboxylase 65 kDa (GAD65), and COMT expression in a nares occlusion model [relative optical density (OD) U/μg total protein]. Expression of TH and GAD65 are reduced 30–40% in the OB on the side of the closed nares, reflecting activity dependence of these proteins. COMT expression is identical in open and closed nares.

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    Figure 3.

    Comparison of COMT and TH expression in different strains/transgenic mice [relative optical density (OD) U/μg total protein]. A, Expression levels of COMT (black line) and TH (gray line) between lines of mice varied as much as 2.5×. B, Regression of relative COMT (ordinate) and TH (abscissa) between lines of mice.

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    Figure 4.

    In vitro FSCV of DA release and metabolism in OB slices. A, FSCV detection of DA release evoked by optical stimulation (blue bar) of ChR2-expressing SACs. Plot shows current in pseudo-color versus applied potential (ordinate) and time (abscissa). Oxidative DA current in green. Inset, The voltammogram at the DA voltage potential. B, The COMT inhibitor, tolcapone (10 μm), increases DA signal. C, FSCV DA oxidation current from (A) control and from (B) tolcapone. D, Average graph of the peak DA oxidative current in control conditions and in the presence of tolcapone. E, FSCV detection of DA release evoked by optical stimulation (blue bar) of ChR2-expressing SACs. F, The DAT inhibitor, GBR12909 (5 μm), does not alter the DA signal. G, FSCV DA oxidation current from (A) control and from (B) GBR12909. H, Average graph of the peak DA oxidative current in control conditions and in the presence of GBR12909.

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The Journal of Neuroscience: 36 (29)
Journal of Neuroscience
Vol. 36, Issue 29
20 Jul 2016
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Subsecond Regulation of Synaptically Released Dopamine by COMT in the Olfactory Bulb
Renee Cockerham, Shaolin Liu, Roger Cachope, Emi Kiyokage, Joseph F. Cheer, Michael T. Shipley, Adam C. Puche
Journal of Neuroscience 20 July 2016, 36 (29) 7779-7785; DOI: 10.1523/JNEUROSCI.0658-16.2016

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Subsecond Regulation of Synaptically Released Dopamine by COMT in the Olfactory Bulb
Renee Cockerham, Shaolin Liu, Roger Cachope, Emi Kiyokage, Joseph F. Cheer, Michael T. Shipley, Adam C. Puche
Journal of Neuroscience 20 July 2016, 36 (29) 7779-7785; DOI: 10.1523/JNEUROSCI.0658-16.2016
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Keywords

  • COMT
  • dopamine
  • enzymatic
  • olfactory
  • transporter
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