title: Generation of Oxtr cDNA(HA)-Ires-Cre Mice for Gene Expression in an Oxytocin Receptor Specific Manner.
abstract: The neurohypophysial hormone oxytocin (OXT) and its receptor (OXTR) have critical roles in the regulation of pro-social behaviors, including social recognition, pair bonding, parental behavior, and stress-related responses. Supporting this hypothesis, a portion of patients suffering from autism spectrum disorder have mutations, such as single nucleotide polymorphisms, or epigenetic modifications in their OXTR gene. We previously reported that OXTR-deficient mice exhibit pervasive social deficits, indicating the critical role of OXTR in social behaviors. In the present study, we generated Oxtr cDNA(HA)-Ires-Cre knock-in mice, expressing both OXTR and Cre recombinase under the control of the endogenous Oxtr promoter. Knock-in cassette of Oxtr cDNA(HA)-Ires-Cre consisted of Oxtr cDNA tagged with the hemagglutinin epitope at the 3' end (Oxtr cDNA(HA)), internal ribosomal entry site (Ires), and Cre. Cre was expressed in the uterus, mammary gland, kidney, and brain of Oxtr cDNA(HA)-Ires-Cre knock-in mice. Furthermore, the distribution of Cre in the brain was similar to that observed in Oxtr-Venus fluorescent protein expressing mice (Oxtr-Venus), another animal model previously generated by our group. Social behavior of Oxtr cDNA(HA)-Ires-Cre knock-in mice was similar to that of wild-type animals. We demonstrated that this construct is expressed in OXTR-expressing neurons specifically after an infection with the recombinant adeno-associated virus carrying the flip-excision switch vector. Using this system, we showed the transport of the wheat-germ agglutinin tracing molecule from the OXTR-expressing neurons to the innervated neurons in knock-in mice. This study might contribute to the monosynaptic analysis of neuronal circuits and to the optogenetic analysis of neurons expressing OXTR.
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Shizu
authors.initials: S
authors.lastname: Hidema
authors.affiliation: Department of Animal Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Fukuda
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Hiraoka
authors.affiliation: Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mizukami
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Ryotaro
authors.initials: R
authors.lastname: Hayashi
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Ayano
authors.initials: A
authors.lastname: Otsuka
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Miyazaki
authors.affiliation: Department of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Nishimori
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008425:Maternal Behavior; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009043:Motor Activity / Q000235:genetics; D009474:Neurons / Q000378:metabolism; D011247:Pregnancy; D011401:Promoter Regions, Genetic / Q000235:genetics*; D018045:Receptors, Oxytocin / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012919:Social Behavior; D014909:Wheat Germ Agglutinins / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D018076:DNA, Complementary; D019426:Integrases; D008164:Luminescent Proteins; D018045:Receptors, Oxytocin; D014909:Wheat Germ Agglutinins; D000097573:Red Fluorescent Protein; C576074:OXTR protein, mouse; C045073:Cre recombinase
keywords: CRE RECOMBINASE; FLIP-EXCISION (FLEX) SWITCH; OXYTOCIN RECEPTOR; PRO-SOCIAL BEHAVIOR