Comments and Questions:

Comments:

For several decades the vast majority of brain slice physiologists have relied upon aprotective cutting method for preparing healthy brain slices from juvenile and adolescent animals. The author used “protective recovering” method which reduces swelling and damage in superficial layers of the slices and improved the success rate for targeted patch clamp recordings, and gave many advices to select the opsin and report protein for using optical genetic technology in recordings. They provide more options to extend the versatility of our methods for particularly challenging applications.

Questions:

1. Both PH and osmotic pressure in intracellular pipette solution are lower than artificial cerebrospinal fluid. Is this used to mimic the physiological conditions? Are there any other reasons?

2. What are the effects of HEPES, sodium ascorbate, thiourea, sodium pyruvate and and NMDG in artificial cerebrospinal fluid?

3. Why did the author use artificial cerebrospinal fluid at room temperature during transcardial perfusion?

4. Normally, high potassium can trigger depolarization of neurons, but why did not it work in the result A in Figure 4?

5. Why did the author insert P2A between ChETA and tdTomato instead of between ChR2 and tdTomato?

6. What are the meanings of 0.31, 0.64 and 1.00 in Figure7D?

7. Is averdin the best anesthetic drug in this work?

8. Could the protective recovering method be used to dramatically improving the health of magnocellular endocrine neurons in brain slices?

2017年12月25日

2017-12- 19 Literature Analysis

Jonathan T. Ting, Tanya L. Daigle, Qian Chen, and Guoping Feng. Acute brain slice methods for adult and aging animals: application of targeted patch clampanalysis and optogenetics.Methods Mol Biol. 2014 ; 1183: 221–242. doi:10.1007/978-1-4939-1096-0_14. PMID: 25023312, Presented by Tongli. Edited by Xiaoran Wang.

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