The following murine models were employed in this study: (a) C57BL/6J; (b) complement component C3‐targeted mutations, B6;129S4‐C3tm1Crr/J mice (C3−/−) (Wessels et al., 1995 (link)); (c) complement component C1q‐targeted mutation, B6(Cg)‐ C1qtm1c(EUCOMM)Wtsi (C1q−/−) (Fonseca et al., 2017 (link)). Mice used as homozygotes and genotypes were confirmed according to the appropriate genotyping PCR protocol (Jackson Laboratories). All mice were infected at 5–6 weeks of age and were euthanized by 8 weeks old. Wild‐type (WT) C57BL/6J animals were acquired from Jackson Laboratory and were age and sex‐matched to the genetically modified groups.
C57bl 6j animals
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C57BL/6J animals are a commonly used inbred strain of laboratory mice. They are known for their black coat color and are widely utilized in biomedical research due to their well-characterized genetic background.
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14 protocols using c57bl 6j animals
1
Murine Models in Complement Research
2
Transgenic Mouse Strain Generation
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C57BL/6J animals were purchased from Jackson Laboratory, Slac Laboratory Animal (Shanghai) and bred in house. Esr1-2a-Cre (Esr1Cre, #017913) and Vglut2-Ires-Cre (Vglut2Cre, #012569) were purchased from the Jackson Laboratory. The Vgat-Cre (VgatCre) line was previously generated68 (link). All Cre lines were bred onto C57BL/6J background for at least one generation. Animals were housed in the Institute of Neuroscience animal facility on 12 h:12 h light/dark cycle with food and water ad libitum. Only heterozygous animals of Cre alleles were used in the study. All experimental protocols were approved by the Animal Care and Use Committee of the Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China (IACUC No. NA-016-2016).
3
Transcriptomics of Mouse Lifespan
4
Pharmacogenetic Ablation of Neurogenesis
5
In-House Breeding and Care of C57/BL6J Mice
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C57/BL6J animals were purchased from Jackson Laboratory (Bar Harbor, ME) and mice were bred in‐house for work done at Università Cattolica del Sacro Cuore. Mice were housed, n ≤ 5 per cage, with food and water ad libitum. Mice were closely monitored for health and overall well‐being daily by veterinary staff and the investigators. Animal maintenance, all surgical procedures and experiments were performed in accordance with the US National Institutes of Health (NIH) guidelines and were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Texas Medical Branch. For work done at Università Cattolica del Sacro Cuore, all animal procedures were approved by the Ethics Committee of Università Cattolica del Sacro Cuore and complied with the Italian Ministry of Health guidelines and with national laws (Legislative Decree 116/1992) and European Union guidelines on animal research (86/609/EEC).
6
Modeling Lipoprotein Metabolism in Mice
7
Forelimb Reaching Task for Spinal Cord Injury
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All animal experiments and procedures were approved by the Weill Cornell Medicine Institutional Animal Care and Use Committee (protocol # 2015-0042) on May 12, 2018. All mice were housed on a 12-hour light/dark cycle from 9 a.m. to 9 p.m. at 25°C with free access to food and water. Twenty-four male and female C57BL/6J animals (8–12 weeks old) were purchased from Jackson Laboratory. For forelimb reaching task, animals were food restricted to 80–90% of their free-feeding bodyweight. Cervical spinal cord dorsal column lesion was used as a model of incomplete injury. Twelve mice used in this study performed the recessed single pellet-reaching task, the rotarod test, and the open field test. In each behavior test, six animals were injected with either nicotinic inhibitors or saline control. Experimental design is shown in Figure 1
8
Generating Obese Mouse Models for Metabolic Research
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Animal housing conditions and research protocols were approved by the Amgen Institutional Animal Care and Use Committee (IACUC). Mice were housed in a specified-pathogen free, AAALAC, Intl-accredited facility in ventilated microisolators. Procedures and housing rooms are positively pressured and regulated on a 12:12 dark:light cycle. All animals received reverse-osmosis purified water ad libitum via an automatic watering system. FGFR4 KO mice and WT littermates were generated as described earlier (Ge et al., 2014 (link)). C57BL/6J animals (The Jackson Laboratory) were singly housed and fed standard chow (2020 × Teklad global soy protein-free extruded rodent diet; Harlan). For diet-induced obese (DIO) animal studies, 16–18-week-old C57BL/6J male mice fed a 60 kcal% fat diet (D12492, Research Diets) for 10 weeks were purchased from the Jackson Laboratory.
Cholecystectomy was performed on 16–18 week old DIO animals at the Jackson Laboratory. In brief, after animals were treated with surgical anesthesia, excision of the gallbladder and ligature of the cystic duct and attached artery were performed. Animals were monitored for recovery before shipment.
For studies with protein injection, mice were intraperitoneally (i.p.) injected with recombinant FGF19 or human FGF21 protein (at 1 mg/kg body weight in 0.2 ml PBS) or an equal volume of PBS as a control.
Cholecystectomy was performed on 16–18 week old DIO animals at the Jackson Laboratory. In brief, after animals were treated with surgical anesthesia, excision of the gallbladder and ligature of the cystic duct and attached artery were performed. Animals were monitored for recovery before shipment.
For studies with protein injection, mice were intraperitoneally (i.p.) injected with recombinant FGF19 or human FGF21 protein (at 1 mg/kg body weight in 0.2 ml PBS) or an equal volume of PBS as a control.
9
Murine Models for Complement Deficiency
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The following murine models were employed in this study: (i) C57BL/6J mice; (ii) for complement component C3-targeted mutations, B6;129S4-C3tm1Crr/J mice (C3−/−) (4 (link)), (iii) for complement component C9-targeted mutation, B6N(Cg)-C9tm1.1(KOMP)Vlcg/J (C9−/−) mice (51 (link)); and (iv) for complement receptor 1/2 (cr2−/−)-targeted mutation, B6.129S7(NOD)-CR2tm1Hmo/J (CR1/2−/−) mice (24 (link)). Mice were used as homozygotes, and genotypes were confirmed according to the appropriate genotyping PCR protocol (Jackson Laboratories). All infected mice were utilized at 5 to 7 weeks of age and were euthanized by 11 weeks old. Wild-type (WT) C57BL/6J animals were acquired from Jackson Laboratory and were age and sex matched to the genetically modified groups.
10
Transcriptomics of Mouse Lifespan
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C57BL/6J animals were obtained from Jackson Laboratory. 5 mice at 5 different age points (8wk = human 20yr, 26wk = human 34yr, 60wk = human 60yr, 78wk = human 65yr, and 104wk = human 70yr) from across mouse lifespan were sacrificed to harvest 11 types of tissues including brain, hypothalamus, lung, bone marrow, gastric muscle, liver, kidney, heart, inguinal white adipose tissue (iWAT), epididymal white adipose tissue (eWAT), and brown adipose tissue (BAT) (Dutta and Sengupta, 2016 (link)). A total of 275 tissue samples were prepared for RNA sequencing.
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