cre-reporter expressing eyfp ai3 mice  (Jackson Laboratory)

Journal: Scientific Reports

Article Title: A platform for stereological quantitative analysis of the brain-wide distribution of type-specific neurons

doi: 10.1038/s41598-017-14699-w

Figure Lengend Snippet: Distribution of EYFP-labelled interneurons and specific co-localization with SOM. ( a ) Gene elements of the Ai3 Cre-reporter mouse and SOM-Cre-line mouse used in the experiments. ( b ) Representative examples of specific co-localization of EYFP-labelled neurons (top, green) from a SOM-IRES-Cre:Ai3-EYFP mouse with immunofluorescence for somatostatin (middle, red). Merged images are shown at the bottom. Scale bars, 100 µm. Nearly all of the EYFP-labelled neurons from SOM-IRES-Cre:Ai3-EYFP mice co-localized with somatostatin in MOp, HPF and CP. Abbreviations: MOp, Primary motor area; HPF, Hippocampal formation and CP, Caudoputamen. SOM-positive rates in MOp, HPF and CP were 74.27 ± 1.31% (n = 3), 80.80 ± 1.39% (n = 3) and 77.53 ± 2.19% (n = 3), respectively.

Article Snippet: The Cre-reporter expressing EYFP Ai3 mice (JAX no. 007903) were purchased from Jackson Laboratories.

Techniques: Immunofluorescence

Journal: Scientific Reports

Article Title: A platform for stereological quantitative analysis of the brain-wide distribution of type-specific neurons

doi: 10.1038/s41598-017-14699-w

Figure Lengend Snippet: Whole-brain imaging of a SOM-IRES-Cre:Ai3-EYFP mouse brain. Images of EYFP-labelled neurons ( a ) and PI-stained cytoarchitecture ( b ) in the hippocampal coronal plane. ( c ) Enlarged views of white rectangular boxes indicated in ( a ) and ( b ) and their merged image. Image size is 900 × 400 μm. Projection thickness in the EYFP channel was 20 μm. ( d ) Sagittal reconstruction of maximum intensity projections of the SOM-IRES-Cre:Ai3-EYFP mouse brain. The projection thickness shown in ( d ) is 20 μm. The insets (d1)–(d5) are enlarged views of white square boxes shown in ( d ). The sizes of (d1)–(d5) are 400 × 400 × 20 μm. Scale bar: ( a ) and ( b ) 1 mm, ( c ) 25 μm, (c1)–(c3) 50 μm, ( d ) 1 mm and (d1)–(d5) 100 μm.

Article Snippet: The Cre-reporter expressing EYFP Ai3 mice (JAX no. 007903) were purchased from Jackson Laboratories.

Techniques: Imaging, Staining

Journal: Scientific Reports

Article Title: A platform for stereological quantitative analysis of the brain-wide distribution of type-specific neurons

doi: 10.1038/s41598-017-14699-w

Figure Lengend Snippet: Stereological cell counting accuracy using the NeuroGPS algorithm. ( a ) Automatically locating the neurons in an SC data cube from a SOM-IRES-Cre:Ai3-EYFP mouse brain dataset using NeuroGPS. Grey represents EYFP-labelled somas in the raw data, and red dots indicate identified soma centres using the NeuroGPS algorithm. Overlapping grey somas and red dots indicate the correct identification of neurons. Yellow arrows represent erroneous commission. ( b ) The 100 μm-thick Z-projection image of the data cube in ( a ). Red arrows represent some disjunctive cells in the z-direction in ( a ) that partially overlap in ( b ). ( c ) Comparison between stereological and planar cell counting in 3D data cubes and their own z-projection images, respectively. ( d ) Accuracy of automated stereological cell counting in the regions of SOM-IRES-Cre:Ai3-EYFP mice brains (n = 3). Abbreviations: MOs, Secondary motor area; LS, Lateral septal nucleus; BST, Bed nuclei of the stria terminalis; cc, Corpus callosum; PIR, Piriform area; SSp, Primary somatosensory area; SSs, Supplemental somatosensory area; BLA, Basolateral amygdala nucleus; LA, Lateral amygdala nucleus; VISp, Primary visual area; VISl, Lateral visual areas; HY, Hypothalamus; CEA, Central amygdala nucleus; SC, Superior colliculus; CB, Cerebellum.

Article Snippet: The Cre-reporter expressing EYFP Ai3 mice (JAX no. 007903) were purchased from Jackson Laboratories.

Techniques: Cell Counting, Comparison

Journal: Scientific Reports

Article Title: A platform for stereological quantitative analysis of the brain-wide distribution of type-specific neurons

doi: 10.1038/s41598-017-14699-w

Figure Lengend Snippet: Brain-wide distribution of SOM-expressing neurons. ( a ) 3D volume rendering of the whole-brain dataset of a SOM-IRES-Cre:Ai3-EYFP mouse. Green represents the brain-wide expression of SOM. Dashed lines indicate the locations of images shown in ( b – g ). ( b – g ) A series of representative coronal images showing the distribution of SOM neurons in selected brain regions at a resampling resolution of 1 × 1 × 2 μm. The insets show corresponding partial enlarged images in ( b – g ) at the original resolution of 0.32 × 0.32 × 2 μm. Scale bar: ( b – g ) 1 mm and (insets) 50 μm.

Article Snippet: The Cre-reporter expressing EYFP Ai3 mice (JAX no. 007903) were purchased from Jackson Laboratories.

Techniques: Expressing

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: Specific EYFP-expression of CRH-IRES-Cre;Ai3 mouse brain. (A–J) Representative photomicrographs depicting CRH neurons expressing EYFP (green), CRH immunostaining (red) and their merged images in MOB, CEA, PVH, B, ECU, IO, MPN, PVT, PG, and TRN, respectively, in a CRH-IRES-Cre;Ai3 mouse brain. The arrowheads show EYFP and CRH double-positive neurons. Scale bar: 40 μm. Abbreviations: main olfactory bulb (MOB), central amygdalar nucleus (CEA), paraventricular hypothalamic nucleus (PVH), Barrington’s nucleus (B), external cuneate nucleus (ECU), inferior olivary complex (IO), medial preoptic nucleus (MPN), paraventricular nucleus of the thalamus (PVT), pontine gray (PG), and tegmental reticular nucleus (TRN).

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Expressing, Immunostaining

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: The brain-wide distribution of CRH neurons imaged by BPS. (A) Three-dimensional rendering of EYFP-labeled CRH neurons. Resampling at 2 μm × 2 μm × 2 μm. (B) Sagittal reconstruction of the maximum intensity projection at the location indicated by a dot dash line in (A) in the CRH-IRES-Cre;Ai3 mouse brain. Projection thickness was 100 μm. (C) The 50-μm-thick maximum intensity projections of the coronal slices indicated by dash lines in (B) , respectively. (D) Enlarged views of EYFP-labeled CRH neurons of the area indicated with text annotations and white boxes in (C). Images of MOs and HPF were the maximum intensity projections of 50 μm, reflecting the sparse distribution of CRH neurons. Images of PIR, ACB, PVH, B, and IO were original data of a single slice. (E) Original images of EYFP-labeled CRH neurons, PI-stained cytoarchitecture and the merge of enlarged views of the area indicated by a red arrowhead and a white box in CEA in (c3). Scale bars: (A,B) 1 mm, (C) 1 mm, and (D) 30 μm. Secondary motor area (MOs), piriform area (PIR), nucleus accumbens (ACB), PVH, CEA, hippocampal formation (HPF), B, and IO.

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Labeling, Staining

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: Locating EYFP-labeled CRH neurons in the whole CRH-IRES-Cre;Ai3 mouse brain. (A) Locating neurons in 100 μm typical image stacks, presented in maximum intensity projection. (B) Three-dimensional reconstruction and (C,D) projection views of the data cubes indicated as white boxes in (A) , respectively. Gray and green signals represent CRH neuronal somas. Red dots represent the recognized centers of CRH neurons. The white, red, blue and orange arrowheads indicated the axial-overlapping neurons, low intensity neurons, error recognition neurons and misidentification neurons, respectively. (E) Counting accuracy. Blue and orange represent the recall and precision ( n = 12), respectively. Each data cube was randomly selected at a size of 300 μm × 300 μm × 300 μm. Scale bars: (A) 1 mm and (C,D) 50 μm.

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Labeling

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: The segmentation of anatomical regions. (A) A 10-μm maximum intensity projection of representative PI-stained coronal slices in the same mouse brain. The cyan lines represent the segmented results using the ANTS tool. (B) EYFP-labeled CRH neurons and PI-stained cytoarchitecture of the enlarged view of the area indicated by a green rectangular box in (A) . Green represents the 100 μm maximum intensity projection of EYFP-labeled CRH neurons. Red represents the 2 μm original data of PI-stained cytoarchitecture. White lines represent the laminated contours of the cortex. (C) Quantitative statistics of EYFP-labeled CRH neurons in different layers in MOp, MOs, VIS, AUD, SSp and SSs. Scale bar: (A) 1 mm, (B) 100 μm. Abbreviations: olfactory areas (OLF), pallidum (PAL), striatum (STR), hypothalamus (HY), cortical subplate (CTXsp), hippocampal formation (HPF), thalamus (TH), midbrain (MB), pons (P), medulla (MY), cerebellum (CB), primary motor area (MOp), secondary motor area (MOs), visual area (VIS), auditory area (AUD), primary somatosensory area (SSp), and supplemental somatosensory area (SSs).

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Staining, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: Quantitative statistics of the distribution of EYFP-labeled CRH neurons in the CRH-IRES-Cre;Ai3 mouse brain. (A) Recognized positions of EYFP-labeled neurons in a CRH-IRES-Cre;Ai3 mouse brain. Different color points represent the soma centers of CRH neurons in different regions. (B) Cell densities of EYFP-labeled CRH neurons in 12 brain regions. Color definitions are the same as those shown in (A) . (C) Cell density and nuclei distance of EYFP-labeled CRH neurons in the sub-regions of the above brain regions.

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Labeling

Journal: Frontiers in Neuroanatomy

Article Title: A Quantitative Analysis of the Distribution of CRH Neurons in Whole Mouse Brain

doi: 10.3389/fnana.2017.00063

Figure Lengend Snippet: Analyzing soma morphology of EYFP-labeled CRH neurons in eleven discrete brain areas. (A) EYFP-labeled CRH neurons in 12 discrete brain areas. Each data cube was randomly selected at the size of 200 μm × 200 μm × 200 μm. Scale bar: 50 μm. (B) Extracting the morphological features of CRH neuronal somas using PCA transformation. Two normalized main features were acquired after PCA on the six morphological parameters. The cyan lines surrounding the most concentrated neurons with similar soma morphology account for 80% of the total neurons in each region. (C) SDs of both main features in the main regions except CB. (D) An ANOVA was conducted to analyze both of the main features. The light and dark colors represent P < 0.05 and P ≥ 0.05, respectively. Primary somatosensory area (SSp), MOB, Ammon’s horn (CA), endopiriform nucleus, dorsal part (EPd), CEA, bed nuclei of the stria terminalis (BST), central medial nucleus of the thalamus (CM), PVH, periaqueductal gray (PAG), PG, IO, and hemispheric regions (HEM).

Article Snippet: CRH-Cre transgenic mice ( ) and EYFP Cre reporter Ai3 mice ( ) were obtained from the Josh Huang Lab (CSHL, Cold Spring Harbor, NY, United States) and Jackson Laboratories (JAX Mice stock number: 007903), respectively.

Techniques: Labeling, Transformation Assay

Journal: Nature medicine

Article Title: Neuronal Targets of Mutant Huntingtin Genetic Reduction to Ameliorate Huntington’s Disease Pathogenesis in Mice

doi: 10.1038/nm.3514

Figure Lengend Snippet: ( a ) Schematic representation of Cre-mediated genetic reduction of mHTT in BACHD mice. Mutant HTT Exon-1 in the BACHD transgene is flanked by two LoxP sites, so fl-mHTT levels can be genetically reduced in cell types expressing Cre. ( b ) Histochemical staining for β-galactosidase to illustrate the Cre-mediated recombination patterns in double transgenic mice containing the Rosa26-LacZ reporter and one of the Cre mouse lines. Black arrows point to the ventral hippocampus, where Cre recombinase is expressed in Emx1-Cre and Emx1-Cre/Rgs9-Cre lineages. Scale bar = 2 mm. ( c,d ) Cre recombinase expression patterns in cortex ( c ) and striatum ( d ) of Emx1-Cre, Rgs9-Cre and Emx1-Cre/Rgs9-Cre mice crossed with Ai3 Cre reporter mice (mediating Cre-dependent YFP expression) . Each panel is a composite that shows Cre activity (YFP signals in green) and neuronal staining (NeuN immunofluorescence in red). Scale bar = 100 μm. ( e ) Western blot with 1C2 (specific to expanded polyglutamine epitope) to detect mHTT but not WT murine Htt in BACHD mice. Subsequent quantitation reveals fl-mHTT protein levels in the cortex, striatum, and cerebellum of BACHD, BE, BR and BER mice ( n = 4 per genotype; *** P < 0.001, ** P < 0.01, * P < 0.05, one-way ANOVA followed by LSD post hoc test). ( f ) In situ hybridization with human m HTT exon-1 specific riboprobe reveals selective reduction of mHTT transcripts in the cortex, striatum or both regions in BE, BR and BER brains, respectively. Scale bar = 1 mm.

Article Snippet: Additionally, Emx1-Cre/Rgs9-Cre double transgenic mice were mated with Ai3 (Gt(ROSA)26Sor tm3(CAG-EYFP)Hze /J) Cre reporter mice from the Jackson Laboratory (JAX007903) .

Techniques: Mutagenesis, Expressing, Staining, Transgenic Assay, Activity Assay, Immunofluorescence, Western Blot, Quantitation Assay, In Situ Hybridization