2020 Fiscal Year Annual Research Report
To figure our the role of CeA CRF neurons in anxiety-promote wakefulness
| Project/Area Number |
20J11041
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| Research Institution | Nagoya University |
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Principal Investigator |
洪 啓栄 名古屋大学, 医学系研究科, 特別研究員(DC2)
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| Project Period (FY) |
2020-04-24 – 2022-03-31
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| Keywords | anxiety / wakefulness / CRF / sleep |
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| Outline of Annual Research Achievements |
The higher level of anxiety causes insomnia, and insomnia also promote anxiety. This feedforward circle is thought to contribute in depression and mental disease. Understanding the neural mechanism of this circle helps to treat patients. I confirmed that corticotropin-releasing factor (CRF) producing neurons in the central amygdala (CeA) can promote anxiety and wakefulness, so I would like to figure out the neural circuit and molecular involved in anxiety and wakefulness.
Firstly, I identified the projection areas, such as the dorsal raphe and the lateral hypothalamus, from CeA CRF neurons by solely expression of fluorescence protein. These results help to understand the neuronal circuit that involved in anxiety-promoting wakefulness.
Next, the neuronal activity of CeA CRF neurons were successfully recorded under anxiety behavior and sleep/wakefulness by micro-endoscope. I had not finished the analysis, but this result would help to understand that the anxiety- and wakefulness-promoting neurons are the same neurons or not.
Thirdly, I successfully recorded several wake-promoting molecular which is able to activate CeA CRF neurons in vitro electrophysiological recording. This proofed that CeA CRF neurons have a role in the anxiety-wakefulness feedforward circle.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The experiment according to research plan submitted previously are performed well as my prediction. I found out several good results and reach numerous goals.
1. Successfully confirmation the role of central amygdala (CeA) corticotropin releasing factor (CRF) neurons in anxiety and wakefulness by designer receptors exclusively activated by designer drugs (DREADD).2. Successfully recording neural activity of (CRF) neurons in the central amygdala (CeA) during sleep and anxiety situation by micro-endoscope (nVista). 3. I found out that several wake-promoting-substances such as dopamine, serotonin and CRF itself activate CeA CRF neurons in vitro electrophysiological assay. 4. I also prepared the mouse (LSL-Cas9), adeno-associated virus (AAV) for in vivo genome editing via CRISPR/Cas9.
Based on above, I judged that his research is going well as expected.
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| Strategy for Future Research Activity |
In the final year of this project, I would focus on the in vivo and in vitro calcium imaging in wakefulness and anxiety, and neurotransmitter which is involved in anxiety and wakefulness.
Subsequently, successfully recorded in vivo neuronal activity during anxiety situation and wakefulness, I am developing auto-analysis system in Matlab and python. I would like to know the wake-activating neurons are also activated during anxiety or not. The preliminary results showed CeA CRF neurons are heterogenous in wake and anxiety. Since this, I need to record in vitro calcium imaging for screening the molecular involved in wakefulness and anxiety. Eventually, I use LSL-Cas9 mice and gRNA to specifically knockout the targeted neurotransmitters in CeA CRF neurons, and mouse would be examined the change of wakefulness and anxiety.
After all these jobs, I plan to make a story how CeA CRF neurons modify wakefulness.
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