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Hirokazu Hirai
Gunma University Graduate School of Medicine
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0721-4293
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GABAergic interneurons play a critical role in tuning neural networks in the central nervous system, and their defects cause neuropsychiatric disorders. Currently, the mDlx enhancer is solely used for adeno-associated virus (AAV) vector-mediated transgene delivery into cortical interneurons. Here, we developed a new inhibitory neuron-specific promoter of a 2.5-kb length from a genomic region of a mouse upstream of exon 1 of a gene encoding glutamic acid decarboxylase (GAD) 65 (mGAD65 promoter). Intravenous infusion of blood-brain barrier-penetrating AAV-PHPB expressing an enhanced green fluorescent protein under the control of the mGAD65 promoter transduced the whole brain in an inhibitory neuron-specific manner. The specificity and efficiency of the mGAD65 promoter for GABAergic interneurons, which was assessed at the motor cortex, were almost identical to or slightly higher than those of the mDlx enhancer. Immunohistochemical analysis revealed that the mGAD65 promoter preferentially transduced parvalbumin (PV)-expressing interneurons. Notably, the mGAD65 promoter transduced chandelier cells more efficiently than the mDlx enhancer and robustly labeled their synaptic boutons, called the cartridge, targeting the axon initial segments of excitatory pyramidal neurons. These results suggest that the mGAD65 promoter is useful for AAV-mediated targeting and the manipulation of GABAergic neurons with the dominance of cortical PV-expressing neurons, including chandelier cells.
Keywords
Interneuron, inhibitory neuron, Chandelier cell, GAD65, Dlx, GABA, Promoter, AAV, AAV-PHP.B
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Received 08 Sep, 2020
Editorial decision: Major revision
On 08 Sep, 2020
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Received 07 Sep, 2020
Review #1 received
Received 06 Sep, 2020
Reviewer #3 agreed
On 26 Aug, 2020
Reviewer #2 agreed
On 25 Aug, 2020
Reviewers invited
Invitations sent on 23 Aug, 2020
Reviewer #1 agreed
On 23 Aug, 2020
Editor invited
On 13 Aug, 2020
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See the published version of this preprint at Molecular Brain.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Hirokazu Hirai
Gunma University Graduate School of Medicine
Corresponding Author
ORCiD: https://orcid.org/0000-0002-0721-4293
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
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Peer Review Timeline
CURRENT STATUS: Published
View the published article at Molecular Brain.
No community comments so far
Reviewer #1 agreed
On 27 Jan, 2021
Editor assigned
On 26 Jan, 2021
Reviewers invited
Invitations sent on 26 Jan, 2021
Submission checks complete
On 26 Jan, 2021
Editor invited
On 26 Jan, 2021
No comments provided
Review #3 received
Received 13 Jan, 2021
Editorial decision: Minor revision
On 13 Jan, 2021
Review #1 received
Received 04 Jan, 2021
Reviewer #3 agreed
On 30 Dec, 2020
Reviewer #2 agreed
On 29 Dec, 2020
Review #2 received
Received 29 Dec, 2020
Editor assigned
On 28 Dec, 2020
Reviewers invited
Invitations sent on 28 Dec, 2020
Reviewer #1 agreed
On 28 Dec, 2020
Submission checks complete
On 28 Dec, 2020
Editor invited
On 28 Dec, 2020
Version 1
Posted 17 Aug, 2020
No comments provided
Review #2 received
Received 08 Sep, 2020
Editorial decision: Major revision
On 08 Sep, 2020
Review #3 received
Received 07 Sep, 2020
Review #1 received
Received 06 Sep, 2020
Reviewer #3 agreed
On 26 Aug, 2020
Reviewer #2 agreed
On 25 Aug, 2020
Reviewers invited
Invitations sent on 23 Aug, 2020
Reviewer #1 agreed
On 23 Aug, 2020
Editor invited
On 13 Aug, 2020
Editor assigned
On 13 Aug, 2020
Submission checks complete
On 12 Aug, 2020
First submitted
On 07 Aug, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cellular & Molecular Neuroscience
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Molecular Brain.
GABAergic interneurons play a critical role in tuning neural networks in the central nervous system, and their defects cause neuropsychiatric disorders. Currently, the mDlx enhancer is solely used for adeno-associated virus (AAV) vector-mediated transgene delivery into cortical interneurons. Here, we developed a new inhibitory neuron-specific promoter of a 2.5-kb length from a genomic region of a mouse upstream of exon 1 of a gene encoding glutamic acid decarboxylase (GAD) 65 (mGAD65 promoter). Intravenous infusion of blood-brain barrier-penetrating AAV-PHPB expressing an enhanced green fluorescent protein under the control of the mGAD65 promoter transduced the whole brain in an inhibitory neuron-specific manner. The specificity and efficiency of the mGAD65 promoter for GABAergic interneurons, which was assessed at the motor cortex, were almost identical to or slightly higher than those of the mDlx enhancer. Immunohistochemical analysis revealed that the mGAD65 promoter preferentially transduced parvalbumin (PV)-expressing interneurons. Notably, the mGAD65 promoter transduced chandelier cells more efficiently than the mDlx enhancer and robustly labeled their synaptic boutons, called the cartridge, targeting the axon initial segments of excitatory pyramidal neurons. These results suggest that the mGAD65 promoter is useful for AAV-mediated targeting and the manipulation of GABAergic neurons with the dominance of cortical PV-expressing neurons, including chandelier cells.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
This is a list of supplementary files associated with this preprint. Click to download.
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