amigo2 cre transgenic mice  (Jackson Laboratory)

Journal: Journal of Biomedical Science

Article Title: A dorsal CA2 to ventral CA1 circuit contributes to oxytocinergic modulation of long-term social recognition memory

doi: 10.1186/s12929-022-00834-x

Figure Lengend Snippet: Deletion of Oxtr in CA2 excitatory neurons of the mouse hippocampus. A Doubled-labeled confocal immunofluorescent images showing the colocalization of OXTRs (green) and STEP (red) in dCA2 of the mouse hippocampus. The inserts represent high magnification of the boxed area. Scale bar: 50 μm. SO stratum oriens, SP stratum pyramidale, SR stratum radiatum. B PCR screening of tail-derived genomic DNA for selection of Oxtr −/− mice. C Dual-probe FISH images showing the expression of Oxtr mRNA and Amigo2 mRNA in dCA2 of WT and Oxtr −/− mice (counterstained with DAPI, blue). Scale bar, 50 μm. Data was replicated in 4 mice. D Representative images with cresyl violet staining of dCA2 showing that the number of pyramidal neurons was not affected by targeted deletion of Oxtr compared with age-matched WT mice. Group data showing the summary results from 3–4 mice of each group at 12 weeks old. Scale bars: left, 500 μm; right, 50 μm. The total number of animal examined is indicated by n in parenthesis. Error bars represent the SEM

Article Snippet: Homozygous Oxtr -floxed ( Oxtr J/J ; donating investigator: Dr. W. Scott Young III) and heterozygous Amigo2 -Cre transgenic mice (donating investigator: Dr. Steven A. Siegelbaum) were obtained from The Jackson Laboratory.

Techniques: Labeling, Derivative Assay, Selection, Expressing, Staining

Journal: Journal of Biomedical Science

Article Title: A dorsal CA2 to ventral CA1 circuit contributes to oxytocinergic modulation of long-term social recognition memory

doi: 10.1186/s12929-022-00834-x

Figure Lengend Snippet: dCA2 neuronal activity is necessary for the persistence of long-term SRM. A Schematic representation of the experimental design. B Schematic representation of viral injection. Three weeks after stereotaxic injection of AAV 5 -hSyn-DIO-hM4D(Gi)-mCherry or AAV 5 -hSyn-DIO-mCherry into the dCA2 of Amigo2 -Cre mice, mice were subjected to three-chamber paradigm test and long-term SRM retention was tested 7 days after the initial interaction. Mice were intraperitoneally injected with CNO 30 min before the initial interaction. Representative images showing the co-expression of hM4D(Gi)-mCherry and STEP immunoreactivity in dCA2 pyramidal neurons. Scale bar represents 50 μm. C Top, schematic representation of the three-chamber sociability test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing the juvenile stimulus mouse or the empty wire cage. Both hM4D(Gi)/CNO and mCherry/CNO subject mice spent significantly more time interacting with the wire cage containing the juvenile stimulus mouse than the empty wire cage. Bottom right, discrimination index (stimulus minus empty) was similar between groups in the sociability test. D Top, schematic representation of the three-chamber social novelty preference test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 1 mouse, 10 min after the sociability test. Both hM4D(Gi)/CNO and mCherry/CNO subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 1 minus familiar) was comparable between groups in the social novelty preference test. E Top, schematic representation of the three-chamber long-term SRM test. Bottom, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 2 mouse, 7 days after the initial interaction. mCherry/CNO, but not hM4D(Gi)/CNO, subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 2 minus familiar) of hM4D(Gi)/CNO subject mice was significantly less than mCherry/CNO subject mice in 7-day long-term SRM test. The total number of animal examined is indicated by n in parenthesis. Error bars represent the SEM; ** P < 0.01, *** P < 0.001

Article Snippet: Homozygous Oxtr -floxed ( Oxtr J/J ; donating investigator: Dr. W. Scott Young III) and heterozygous Amigo2 -Cre transgenic mice (donating investigator: Dr. Steven A. Siegelbaum) were obtained from The Jackson Laboratory.

Techniques: Activity Assay, Injection, Expressing

Journal: Journal of Biomedical Science

Article Title: A dorsal CA2 to ventral CA1 circuit contributes to oxytocinergic modulation of long-term social recognition memory

doi: 10.1186/s12929-022-00834-x

Figure Lengend Snippet: dCA2 projections to vCA1 is necessary for long-term SRM. A Schematic representation of the experimental design. B Schematic representation of viral injection and CNO administration. Three weeks after stereotaxic injection of AAV 5 -hSyn-DIO-hM4D(Gi)-mCherry into the dCA2 of Amigo2 -Cre mice, mice were subjected to three-chamber paradigm test and long-term SRM retention was tested 7 days after the initial interaction. Mice were bilaterally administered of vehicle (Veh) or CNO into vCA1 20 min before the initial interaction. Representative images showing the co-expression of hM4D(Gi)-mCherry and STEP immunoreactivity in dCA2 pyramidal neurons. Scale bars represent 50 μm. mCherry signals of axonal projections were observed in vCA1. Scale bar represents 100 μm. NeuN neuronal nuclear protein, SO stratum oriens, SP stratum pyramidale, SR stratum radiatum. C Top, schematic representation of the three-chamber sociability test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing the juvenile stimulus mouse or the empty wire cage. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time interacting with the wire cage containing the juvenile stimulus mouse than the empty wire cage. Bottom right, discrimination index (stimulus minus empty) was similar between hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice in the sociability test. D Top, schematic representation of the three-chamber social novelty preference test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 1 mouse, 10 min after the sociability test. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 1 minus familiar) was comparable between hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice in the social novelty preference test. E Top, schematic representation of the three-chamber long-term SRM test. Bottom, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 2 mouse, 7 days after the initial interaction. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 2 minus familiar) of hM4D(Gi)/CNO subject mice was significantly less than hM4D(Gi)/Veh subject mice in 7-day long-term SRM test. The total number of animal examined is indicated by n in parenthesis. Error bars represent the SEM; ** P < 0.01, *** P < 0.001

Article Snippet: Homozygous Oxtr -floxed ( Oxtr J/J ; donating investigator: Dr. W. Scott Young III) and heterozygous Amigo2 -Cre transgenic mice (donating investigator: Dr. Steven A. Siegelbaum) were obtained from The Jackson Laboratory.

Techniques: Injection, Expressing

Journal: Journal of Biomedical Science

Article Title: A dorsal CA2 to ventral CA1 circuit contributes to oxytocinergic modulation of long-term social recognition memory

doi: 10.1186/s12929-022-00834-x

Figure Lengend Snippet: dCA2 projections to dLS is not involved in long-term SRM. A Schematic representation of the experimental design. B Schematic representation of viral injection. Three weeks after stereotaxic injection of AAV 5 -hSyn-DIO-hM4D(Gi)-mCherry into the dCA2 of Amigo2 -Cre mice, mice were subjected to three-chamber paradigm test and long-term SRM retention was assessed 7 days after the initial interaction. Mice were bilaterally administered of vehicle (Veh) or CNO into LS 20 min before the initial interaction. Representative images showing the co-expression of hM4D(Gi)-mCherry and STEP immunoreactivity in dCA2 pyramidal neurons. Scale bars represent 50 μm. mCherry signals of axonal projections were observed in dLS. Scale bar represents 100 μm. NeuN neuronal nuclear protein. C Top, schematic representation of the three-chamber sociability test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing the juvenile stimulus mouse or the empty wire cage. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time interacting with the wire cage containing the juvenile stimulus mouse than the empty wire cage. Bottom right, discrimination index (stimulus minus empty) was similar between hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice in the sociability test. D Top, schematic representation of the three-chamber social novelty preference test. Bottom left, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 1 mouse, 10 min after the sociability test. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 1 minus familiar) was comparable between hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice in the social novelty preference test. E Top, schematic representation of the three-chamber long-term SRM test. Bottom, time spent by the subject mouse in sniffing directed at the wire cage containing a familiar mouse or a novel 2 mouse, 7 days after the initial interaction. Both hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice spent significantly more time sniffing the cage containing the novel mouse than the familiar mouse. Bottom right, discrimination index (novel 2 minus familiar) was similar between hM4D(Gi)/Veh and hM4D(Gi)/CNO subject mice in 7-day long-term SRM test. The total number of animal examined is indicated by n in parenthesis. Error bars represent the SEM; * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Homozygous Oxtr -floxed ( Oxtr J/J ; donating investigator: Dr. W. Scott Young III) and heterozygous Amigo2 -Cre transgenic mice (donating investigator: Dr. Steven A. Siegelbaum) were obtained from The Jackson Laboratory.

Techniques: Injection, Expressing