Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X
Dynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indi...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2022-10-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/76691 |
| _version_ | 1811186679197728768 |
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| author | Jinli Geng Yingjun Tang Zhen Yu Yunming Gao Wenxiang Li Yitong Lu Bo Wang Huiming Zhou Ping Li Nan Liu Ping Wang Yubo Fan Yaxiong Yang Zengcai V Guo Xiaodong Liu |
| author_facet | Jinli Geng Yingjun Tang Zhen Yu Yunming Gao Wenxiang Li Yitong Lu Bo Wang Huiming Zhou Ping Li Nan Liu Ping Wang Yubo Fan Yaxiong Yang Zengcai V Guo Xiaodong Liu |
| author_sort | Jinli Geng |
| collection | DOAJ |
| description | Dynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indicators that have been widely applied. Here, we demonstrate the performance of GCaMP-X in chronic Ca2+ imaging of cortical neurons, where GCaMP-X by design is to eliminate the unwanted interactions between the conventional GCaMP and endogenous (apo)CaM-binding proteins. By expressing in adult mice at high levels over an extended time frame, GCaMP-X showed less damage and improved performance in two-photon imaging of sensory (whisker-deflection) responses or spontaneous Ca2+ fluctuations, in comparison with GCaMP. Chronic Ca2+ imaging of one month or longer was conducted for cultured cortical neurons expressing GCaMP-X, unveiling that spontaneous/local Ca2+ transients progressively developed into autonomous/global Ca2+ oscillations. Along with the morphological indices of neurite length and soma size, the major metrics of oscillatory Ca2+, including rate, amplitude and synchrony were also examined. Dysregulations of both neuritogenesis and Ca2+ oscillations became discernible around 2–3 weeks after virus injection or drug induction to express GCaMP in newborn or mature neurons, which were exacerbated by stronger or prolonged expression of GCaMP. In contrast, neurons expressing GCaMP-X were significantly less damaged or perturbed, altogether highlighting the unique importance of oscillatory Ca2+ to neural development and neuronal health. In summary, GCaMP-X provides a viable solution for Ca2+ imaging applications involving long-time and/or high-level expression of Ca2+ probes. |
| first_indexed | 2024-04-11T13:49:50Z |
| format | Article |
| id | doaj.art-5f23f2781b704e41ba0d4a4b9fdef286 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T13:49:50Z |
| publishDate | 2022-10-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-5f23f2781b704e41ba0d4a4b9fdef2862022-12-22T04:20:41ZengeLife Sciences Publications LtdeLife2050-084X2022-10-011110.7554/eLife.76691Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-XJinli Geng0Yingjun Tang1Zhen Yu2Yunming Gao3Wenxiang Li4Yitong Lu5Bo Wang6Huiming Zhou7Ping Li8Nan Liu9Ping Wang10Yubo Fan11Yaxiong Yang12https://orcid.org/0000-0002-3313-6049Zengcai V Guo13https://orcid.org/0000-0002-4140-7961Xiaodong Liu14https://orcid.org/0000-0002-3171-9611Advanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaTsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, School of Medicine, Tsinghua University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, China; Tsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, School of Medicine, Tsinghua University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, ChinaCenter for Life Sciences, School of Life Sciences, Yunnan University, Kunming, ChinaKey Laboratory for Biomedical Engineering of Ministry of Education, Zhejiang University, Hangzhou, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, ChinaTsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, School of Medicine, Tsinghua University, Beijing, ChinaAdvanced Innovation Center for Biomedical Engineering, School of Biological Science and Medical Engineering, School of Engineering Medicine, Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, Beihang University, Beijing, China; X-Laboratory for Ion-Channel Engineering, Beihang University, Beijing, ChinaDynamic Ca2+ signals reflect acute changes in membrane excitability, and also mediate signaling cascades in chronic processes. In both cases, chronic Ca2+ imaging is often desired, but challenged by the cytotoxicity intrinsic to calmodulin (CaM)-based GCaMP, a series of genetically-encoded Ca2+ indicators that have been widely applied. Here, we demonstrate the performance of GCaMP-X in chronic Ca2+ imaging of cortical neurons, where GCaMP-X by design is to eliminate the unwanted interactions between the conventional GCaMP and endogenous (apo)CaM-binding proteins. By expressing in adult mice at high levels over an extended time frame, GCaMP-X showed less damage and improved performance in two-photon imaging of sensory (whisker-deflection) responses or spontaneous Ca2+ fluctuations, in comparison with GCaMP. Chronic Ca2+ imaging of one month or longer was conducted for cultured cortical neurons expressing GCaMP-X, unveiling that spontaneous/local Ca2+ transients progressively developed into autonomous/global Ca2+ oscillations. Along with the morphological indices of neurite length and soma size, the major metrics of oscillatory Ca2+, including rate, amplitude and synchrony were also examined. Dysregulations of both neuritogenesis and Ca2+ oscillations became discernible around 2–3 weeks after virus injection or drug induction to express GCaMP in newborn or mature neurons, which were exacerbated by stronger or prolonged expression of GCaMP. In contrast, neurons expressing GCaMP-X were significantly less damaged or perturbed, altogether highlighting the unique importance of oscillatory Ca2+ to neural development and neuronal health. In summary, GCaMP-X provides a viable solution for Ca2+ imaging applications involving long-time and/or high-level expression of Ca2+ probes.https://elifesciences.org/articles/76691genetically encoded calcium indicatorscortical neuronscalmodulincalcium oscillationsin vivo calcium imagingneurite outgrowth |
| spellingShingle | Jinli Geng Yingjun Tang Zhen Yu Yunming Gao Wenxiang Li Yitong Lu Bo Wang Huiming Zhou Ping Li Nan Liu Ping Wang Yubo Fan Yaxiong Yang Zengcai V Guo Xiaodong Liu Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X eLife genetically encoded calcium indicators cortical neurons calmodulin calcium oscillations in vivo calcium imaging neurite outgrowth |
| title | Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X |
| title_full | Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X |
| title_fullStr | Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X |
| title_full_unstemmed | Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X |
| title_short | Chronic Ca2+ imaging of cortical neurons with long-term expression of GCaMP-X |
| title_sort | chronic ca2 imaging of cortical neurons with long term expression of gcamp x |
| topic | genetically encoded calcium indicators cortical neurons calmodulin calcium oscillations in vivo calcium imaging neurite outgrowth |
| url | https://elifesciences.org/articles/76691 |
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