Improved functional preservation with the protective recovery method:
Optogenetic manipulation of neuronal activity in mature adult brain slices
We explored the potential for optogenetic manipulation of neuronal populations across diverse brain regions in acute brain slices from 8 month old mature adult mice in order to demonstrate the suitability of the protective recovery method for optogenetics-based applications. The improvement in neuronal preservation with the protective recovery method led to greater success in preparing healthy slices from mature animals and in identifying and recording from opsin expressing neurons. These improvements will be helpful for researchers seeking to perform detailed cellular and circuitry analysis of brain function in the same cohorts of animals used in behavioral testing. Acute brain slices can be routinely prepared from mature adult animals upon completion of the behavioral battery of tests, regardless of the age of the animals.
Optogenetic silencing: 8 month old Thy1-NpHR2.0-EYFP transgenic mice
The Thy1-NPHR2.0-EYFP line 2 transgenic mice were developed in the laboratory of Guoping Feng (unpublished). These mice are available for purchase from the Jackson Laboratory (stock #012332).
Optogenetic stimulation: Viral injection of DIO-ChR2-EYFP into dorsal striatum of 8 month old
D2-Cre transgenic mice (viral vector from the laboratory of Karl Deisseroth, Stanford University).