Thy1-YFP-H mice (stock 003782)57 (link), which express yellow fluorescent protein (YFP) in L5 pyramidal neurons, Rosa26-stop-DTR mice (stock 007900 )58 (link), which express a conditional DTR-encoding allele in the ROSA locus, Rag1−/− mice (stock 002216)59 (link), Tnf−/− mice (stock 005540)60 (link), Il6−/− mice (stock 002650) and B6 CD45.1 mice (stock 002014) were purchased from the Jackson Laboratory. Cx3cr1CreER mice35 (link) were obtained from the laboratory of Wen-Biao Gan (New York University). Cx3cr1GFP mice34 (link) were obtained from the laboratory of Dan Littman (New York University). To image dendritic spine plasticity in Cx3cr1CreER/+;R26iDTR/+ and Tnf−/− mice, these mice were crossed to Thy1-YFP-H mice. For systemic immune challenge, mice were administered poly(I:C) (Sigma P1530; by i.p. injection) that was dissolved in DPBS. Mice were group-housed in temperature-controlled rooms on a 12-h light–dark cycle and were randomly assigned to different treatment groups. The group size was determined based on previous studies using the same methodologies28 (link),29 (link),30 (link),31 (link),32 (link). Both male and female mice were used. All mice were maintained at the NYU Skirball Institute specific-pathogen-free animal facility and handled in accordance with the institutional guidelines for animal care and use.
Thy1 yfp h mice
Manufactured by Jackson ImmunoResearch
Sourced in United States
Thy1-YFP-H mice express yellow fluorescent protein (YFP) under the control of the Thy1 promoter, resulting in the expression of YFP in a subset of neurons. This line is useful for visualization and study of neuronal morphology and connectivity.
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Lab products found in correlation
Rosa26 stop dtr mice, by Jackson ImmunoResearch (2 mentions)
B6 cd45.1 mice, by Jackson ImmunoResearch (2 mentions)
Rag1 mice, by Jackson ImmunoResearch (2 mentions)
Tnf mice, by Jackson ImmunoResearch (2 mentions)
Il 6 mice, by Jackson ImmunoResearch (2 mentions)
P1530, by Merck Group (2 mentions)
Cd 1 animals, by Charles River Laboratories (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
Cx3cr1 gfp mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6j, by Jackson ImmunoResearch (1 mentions)
Thy1 gfp m mice, by Jackson ImmunoResearch (1 mentions)
B6 cg tg camk2a tta 1mmay dboj, by Jackson ImmunoResearch (1 mentions)
9 protocols using thy1 yfp h mice
1
Transgenic Mouse Models for Dendritic Spine Imaging
2
Two-Photon Imaging of Thy1-YFP-H Mice
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Thy1-YFP-h mice with a significant presence of expressing yellow fluorescent protein (YFP) in the motor neurons (Feng et al., 2000 (link)) were obtained from the Jackson Laboratory (United States). They were kept under specific-pathogen-free conditions on a 12-h dark/light cycle with food and water ad libitum until use. All animal experiments were approved by the Institutional Animal Care and Use Committee of Hamamatsu University School of Medicine (Permission no.: 2019009) and were performed following the relevant guidelines and regulations. A one year and 6 months-old female heterozygous Thy1-YFP-H mouse was anesthetized using a mixed anesthetic (Kawai et al., 2011 (link)) and subsequently perfusion-fixed with 4% PFA in 0.1-M phosphate buffer (pH 7.4). The brain was isolated and fixed in the same fixative for 1 h at room temperature (approximately 25°C). After several washes with PBS, images were obtained via two-photon microscopy.
3
Retinal Cell Characterization in Mice
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All mice were used in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and with the approval of the University of Colorado Denver Institutional Animal Care and Use Committee. Wild-type CD-1 mice were used for retinal sections at embryonic and postnatal stages. Thy1-YFP-H mice were acquired from Jackson Laboratories (stock #3782; Bar Harbor, ME, USA)62 (link) and maintained by outcrossing to CD-1 animals (Charles River Laboratories, Wilmington, MA, USA). Flatmount stains were done with CD-1 mice or the wild-type littermates of Thy1-YFP-H mice. The Prdm16 staining pattern in retinal flatmounts (below) was equivalent in C57BL/6J mice (Jackson Laboratories, stock #664) and at all ages examined (3–25 weeks; data not shown).
4
Transgenic Mouse Models for Dendritic Spine Imaging
5
Transgenic Mice for DRG Neuron Imaging
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Transgenic mice expressing GCaMP6s in a subset of DRG sensory neurons, Thy1-GCaMP6 slow founder line 3, were generated at New York University School of Medicine. Thy1-YFP-H mice (stock no. 003782) expressing YFP in DRG sensory neurons and Cx3cr1-GFP mice (stock no. 005582) expressing GFP in DRG macrophages were purchased from the Jackson Laboratory.
Two-month to 3-month-old animals of both sexes were used for all the experiments. Mice were group-housed in temperature-controlled rooms on a 12-h light–dark cycle. After vertebral window implantation, mice were housed individually to minimize the risk of window damage. All animal procedures were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee (IACUC) of New York University and Columbia University as consistent with National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals.
Two-month to 3-month-old animals of both sexes were used for all the experiments. Mice were group-housed in temperature-controlled rooms on a 12-h light–dark cycle. After vertebral window implantation, mice were housed individually to minimize the risk of window damage. All animal procedures were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee (IACUC) of New York University and Columbia University as consistent with National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals.
6
In Vivo Imaging of Neuronal Dendritic Spines
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One-to-two-month-old C57BL/6J (The Jackson Laboratory, 000664) and Thy1-YFP-H mice (The Jackson Laboratory, 003782), both male and female, were received from Jackson laboratory, and housed on a 12 dark/12 light cycle at temperatures of 20–26 °C (set point 22.8 °C) with 30–70% humidity (set point 50%) and used for the in vivo experiments described in Fig. 1d–f , Supplementary Figs. 1 –5 and in Fig. 7 and Supplementary Fig. 8 , respectively. The Thy1-YFP-H mice, as previously reported, have YFP expression restricted mainly to subsets of layer V neurons facilitating the identification of dendrites and dendritic spines76 (link).
7
Imaging Dendritic Spines and Calcium Dynamics
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Thy1-YFP-H mice (Stock No.: 003782) were purchased from the Jackson Laboratory and used for dendritic spine imaging experiments. Thy1-GCaMP6slow mice (Founder line 1) were used for Ca2+ imaging experiments (Cichon et al., 2020 (link)). Mice were group-housed in animal facilities at New York University Medical Center and Columbia University Medical Center. All animal procedures were carried out following protocols approved by Institutional Animal Care and Use Committees (IACUC) at New York University and Columbia University as consistent with National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals.
8
In Vivo Imaging of Neuronal Activity
9
Murine Husbandry and Blinding in Animal Studies
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Adult male C57Bl/6 mice (3-months-old) were purchased from the Guangdong Medical Laboratory Animal Center. Adult male Thy1-YFP-H mice (3 months old) were purchased from the Jackson Laboratory (USA). Mice were maintained in a pathogen-free SPFII animal facility in a condition-controlled room (23 ± 1°C, 50 ± 10% humidity). A 12 h light/dark cycle was automatically imposed. Mice were housed in groups of four to five per individually ventilated cages and given access to food and water ad libitum unless otherwise stated in the methods below. Experimenters were blinded to animals’ treatments and sample processing throughout the subsequent experimentation and analyses.
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