カルシウム/カルモジュリン依存性タンパク質キナーゼIαは大脳皮質興奮性神経細胞の放射状移動を制御する

学位の種別: 課程博士審査委員会委員 : (主査)東京大学教授 真鍋 俊也, 東京大学教授 狩野 方伸, 東京大学教授 岩坪 威, 東京大学教授 山岨 達也, 東京大学准教授 田中 輝幸University of Tokyo(東京大学

A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging in vivo

脳の神経活動を可視化する新規マウス系統を開発 --高感度・高速カルシウムセンサーによる神経活動の計測に成功--. 京都大学プレスリリース. 2022-02-15.Genetically encoded calcium indicators (GECIs) are widely used to measure calcium transients in neuronal somata and processes, and their use enables the determination of action potential temporal series in a large population of neurons. Here, we generate a transgenic mouse line expressing a highly sensitive green GECI, G-CaMP9a, in a Flp-dependent manner in excitatory and inhibitory neuronal subpopulations downstream of a strong CAG promoter. Combining this reporter mouse with viral or mouse genetic Flp delivery methods produces a robust and stable G-CaMP9a expression in defined neuronal populations without detectable detrimental effects. In vivo two-photon imaging reveals spontaneous and sensory-evoked calcium transients in excitatory and inhibitory ensembles with cellular resolution. Our results show that this reporter line allows long-term, cell-type-specific investigation of neuronal activity with enhanced resolution in defined populations and facilitates dissecting complex dynamics of neural networks in vivo

Additional file 3: Figure S1. of Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIα pathway during neuronal polarization

Activation of Wnt5a-Ca2+ signaling in immature cultured cortical neurons. Addition of Wnt5a (100 ng/ml, final concentration) to the culture medium evoked a sustained Ca2+ oscillatory response in immature cultured cortical neurons (Cell #1 and Cell #2). Vertical lines indicate the time points of Wnt5a application and of the respective image frames. Scale bar: 5 μm. (PDF 74 kb

Additional file 1: Movie 1. of Facilitation of axon outgrowth via a Wnt5a-CaMKK-CaMKIÎą pathway during neuronal polarization

Fluo-4 Ca2+ imaging of immature migrating cortical neurons during spontaneous Ca2+ activity. Movie 1 (related to Fig. 1c) shows transient Fluo-4 fluorescence signals induced by spontaneous Ca2+ rises, in migrating neurons in CP and IZ as identified by TagRFP expression. Migrating neurons in CP and IZ are magnified from boxed areas in the left panel of the movie. (AVI 3964 kb

Remote control of neural function by X-ray-induced scintillation

Scintillators emit visible luminescence when irradiated with X-rays. Given the unlimited tissue penetration of X-rays, the employment of scintillators could enable remote optogenetic control of neural functions at any depth of the brain. Here we show that a yellow-emitting inorganic scintillator, Ce-doped Gd3(Al,Ga)5O12 (Ce:GAGG), can effectively activate red-shifted excitatory and inhibitory opsins, ChRmine and GtACR1, respectively. Using injectable Ce:GAGG microparticles, we successfully activated and inhibited midbrain dopamine neurons in freely moving mice by X-ray irradiation, producing bidirectional modulation of place preference behavior. Ce:GAGG microparticles are non-cytotoxic and biocompatible, allowing for chronic implantation. Pulsed X-ray irradiation at a clinical dose level is sufficient to elicit behavioral changes without reducing the number of radiosensitive cells in the brain and bone marrow. Thus, scintillator-mediated optogenetics enables minimally invasive, wireless control of cellular functions at any tissue depth in living animals, expanding X-ray applications to functional studies of biology and medicine