A new phenotype identification method with the fluorescent expression in cross-sectioned tails in <i>Thy1</i>-GCaMP6s transgenic mice
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- Kobayashi Shutaro
- Department of Pharmacology, Nihon University School of Dentistry Department of Oral and Maxillofacial Surgery, Nihon University School of Dentistry
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- O’Hashi Kazunori
- Department of Pharmacology, Nihon University School of Dentistry Division of Oral and Craniomaxillofacial Research, Dental Research Center, Nihon University School of Dentistry Department of Mental Disorder Research, National Institute of Neuroscience, National Center of Neurology and Psychiatry
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- Kaneko Keisuke
- Department of Pharmacology, Nihon University School of Dentistry Department of Anesthesiology, Nihon University School of Dentistry
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- Kobayashi Satomi
- Department of Pharmacology, Nihon University School of Dentistry Division of Oral and Craniomaxillofacial Research, Dental Research Center, Nihon University School of Dentistry Department of Biology, Nihon University School of Dentistry
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- Ogisawa Shouhei
- Department of Pharmacology, Nihon University School of Dentistry Department of Oral and Maxillofacial Surgery, Nihon University School of Dentistry
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- Tonogi Morio
- Department of Oral and Maxillofacial Surgery, Nihon University School of Dentistry
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- Fujita Satoshi
- Division of Oral and Craniomaxillofacial Research, Dental Research Center, Nihon University School of Dentistry Department of Biology, Nihon University School of Dentistry
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- Kobayashi Masayuki
- Department of Pharmacology, Nihon University School of Dentistry Division of Oral and Craniomaxillofacial Research, Dental Research Center, Nihon University School of Dentistry Molecular Dynamics Imaging Unit, RIKEN Center for Life Science Technologies
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<p>Purpose: Unless the phenotype of the transgenic mice is distinguishable, genotyping in each mouse is required prior to experiments. This study aimed to establish a new identification method for the phenotype in Thy1-GCaMP6s transgenic mice to reduce the cost and time.</p><p>Methods: Tail biopsies (2 mm) were performed under general anesthesia with isoflurane in 3 to 4-week-old mice. Then, the resected tail was cut again with a sharp razor, and the cross-sections were observed with two-photon microscopy (excitation wavelength = 940 nm). The emitted light was split into green and red light by a dichroic mirror (570 nm) with bandpass filters (495-540 nm for green, 575-645 nm for red).</p><p>Results: Two types of expressed fluorescent pattern were found in the tail tissue: the presence of green fluorescent structures (type 1) and the absence of the structures (type 2). Cortical imaging confirmed that type 1 expressed the cortical GCaMP6s, while type 2 did not.</p><p>Conclusion: These results suggest that observation of the cross-sectioned tail in Thy1-GCaMP6s mice enabled to identify the phenotype within approximately 10 min/mouse, which reduces the cost and time for genotyping.</p>
収録刊行物
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- Ｊｏｕｒｎａｌ ｏｆ Ｏｒａｌ Ｓｃｉｅｎｃｅ
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Ｊｏｕｒｎａｌ ｏｆ Ｏｒａｌ Ｓｃｉｅｎｃｅ 64 (2), 156-160, 2022
日本大学歯学部
