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Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice

The suprachiasmatic nucleus (SCN), the central circadian clock in mammals, is a neural network consisting of various types of GABAergic neurons, which can be differentiated by the co-expression of specific peptides such as vasoactive intestinal peptide (VIP) and arginine vasopressin (AVP). VIP has b...

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Autores principales: Peng, Yubo, Tsuno, Yusuke, Matsui, Ayako, Hiraoka, Yuichi, Tanaka, Kohichi, Horike, Shin-ichi, Daikoku, Takiko, Mieda, Michihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9110775/
https://www.ncbi.nlm.nih.gov/pubmed/35592033
http://dx.doi.org/10.3389/fphys.2022.895633
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author Peng, Yubo
Tsuno, Yusuke
Matsui, Ayako
Hiraoka, Yuichi
Tanaka, Kohichi
Horike, Shin-ichi
Daikoku, Takiko
Mieda, Michihiro
author_facet Peng, Yubo
Tsuno, Yusuke
Matsui, Ayako
Hiraoka, Yuichi
Tanaka, Kohichi
Horike, Shin-ichi
Daikoku, Takiko
Mieda, Michihiro
author_sort Peng, Yubo
collection PubMed
description The suprachiasmatic nucleus (SCN), the central circadian clock in mammals, is a neural network consisting of various types of GABAergic neurons, which can be differentiated by the co-expression of specific peptides such as vasoactive intestinal peptide (VIP) and arginine vasopressin (AVP). VIP has been considered as a critical factor for the circadian rhythmicity and synchronization of individual SCN neurons. However, the precise mechanisms of how VIP neurons regulate SCN circuits remain incompletely understood. Here, we generated Vip ( tTA ) knock-in mice that express tetracycline transactivator (tTA) specifically in VIP neurons by inserting tTA sequence at the start codon of Vip gene. The specific and efficient expression of tTA in VIP neurons was verified using EGFP reporter mice. In addition, combined with Avp-Cre mice, Vip ( tTA ) mice enabled us to simultaneously apply different genetic manipulations to VIP and AVP neurons in the SCN. Immunostaining showed that VIP is expressed at a slightly reduced level in heterozygous Vip ( tTA ) mice but is completely absent in homozygous mice. Consistently, homozygous Vip ( tTA ) mice showed impaired circadian behavioral rhythms similar to those of Vip knockout mice, such as attenuated rhythmicity and shortened circadian period. In contrast, heterozygous mice demonstrated normal circadian behavioral rhythms comparable to wild-type mice. These data suggest that Vip ( tTA ) mice are a valuable genetic tool to express exogenous genes specifically in VIP neurons in both normal and VIP-deficient mice, facilitating the study of VIP neuronal roles in the SCN neural network.
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spelling pubmed-91107752022-05-18 Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice Peng, Yubo Tsuno, Yusuke Matsui, Ayako Hiraoka, Yuichi Tanaka, Kohichi Horike, Shin-ichi Daikoku, Takiko Mieda, Michihiro Front Physiol Physiology The suprachiasmatic nucleus (SCN), the central circadian clock in mammals, is a neural network consisting of various types of GABAergic neurons, which can be differentiated by the co-expression of specific peptides such as vasoactive intestinal peptide (VIP) and arginine vasopressin (AVP). VIP has been considered as a critical factor for the circadian rhythmicity and synchronization of individual SCN neurons. However, the precise mechanisms of how VIP neurons regulate SCN circuits remain incompletely understood. Here, we generated Vip ( tTA ) knock-in mice that express tetracycline transactivator (tTA) specifically in VIP neurons by inserting tTA sequence at the start codon of Vip gene. The specific and efficient expression of tTA in VIP neurons was verified using EGFP reporter mice. In addition, combined with Avp-Cre mice, Vip ( tTA ) mice enabled us to simultaneously apply different genetic manipulations to VIP and AVP neurons in the SCN. Immunostaining showed that VIP is expressed at a slightly reduced level in heterozygous Vip ( tTA ) mice but is completely absent in homozygous mice. Consistently, homozygous Vip ( tTA ) mice showed impaired circadian behavioral rhythms similar to those of Vip knockout mice, such as attenuated rhythmicity and shortened circadian period. In contrast, heterozygous mice demonstrated normal circadian behavioral rhythms comparable to wild-type mice. These data suggest that Vip ( tTA ) mice are a valuable genetic tool to express exogenous genes specifically in VIP neurons in both normal and VIP-deficient mice, facilitating the study of VIP neuronal roles in the SCN neural network. Frontiers Media S.A. 2022-05-03 /pmc/articles/PMC9110775/ /pubmed/35592033 http://dx.doi.org/10.3389/fphys.2022.895633 Text en Copyright © 2022 Peng, Tsuno, Matsui, Hiraoka, Tanaka, Horike, Daikoku and Mieda. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Peng, Yubo
Tsuno, Yusuke
Matsui, Ayako
Hiraoka, Yuichi
Tanaka, Kohichi
Horike, Shin-ichi
Daikoku, Takiko
Mieda, Michihiro
Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title_full Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title_fullStr Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title_full_unstemmed Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title_short Cell Type-Specific Genetic Manipulation and Impaired Circadian Rhythms in Vip ( tTA ) Knock-In Mice
title_sort cell type-specific genetic manipulation and impaired circadian rhythms in vip ( tta ) knock-in mice
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9110775/
https://www.ncbi.nlm.nih.gov/pubmed/35592033
http://dx.doi.org/10.3389/fphys.2022.895633
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