How Information Is Processed Through The Brain, Especially The Diseased Brain

2233 Words9 Pages
The authors in this article are trying to better understand how information is processed in the brain, especially the diseased brain. In order to research this information there first needs to be a precise method of spatio-temporal recording of electrical activity in larger neuronal circuits and individual neurons. The authors hypothesized that fluorescent indicators would be useful tools in recording brain activity because they would allow monitoring of a genetically defined neuronal circuit and they would not require chemical access to visualize and provide optical recording of brain activity. Since the current techniques could not meet the authors’ requirements to study neuronal activity in vivo, they sought out to create a voltage…show more content…
Next the authors made a circularly altered Clover GFP by creating termini at the amino acid positions 145 and 144 and linking its N- and C- termini using the peptide ‘GGTGGS’ between the C-terminal (K239) lysine and the original starting methionine. The authors obtained cpsfGFP-OPT145-144 (abbreviated as cpsfGFP-OPT), which is a circular permutant of a superfolder GFP variant and which contains 7 mutations that are not present in the original GFP superfolder; it was made for the use in a split GFP system. The authors then constructed circularly altered superecliptic pHlurinA227D (cpsepHluorinA227D), creating new termini at the amino acid positions 145 and linking its original N-and C- termini using the peptide GTGGSAS between the C-terminal lysine (K239) and a lysine originally near its N terminus (K4). Then authors constructed ASAP1 using the voltage-sensing domain (VSD) from the Gallus gallus voltage-sensitive phosphatase. The ASAP1 variants were made with the alternate VSDs by inserting cspfGFP-OPT following amino acid G146 (Xenopus laevis), G147 (Danio rerio), or amino acids L203 to G214 (Ciona intestinalis). The authors isolated the VSD from the VSP by cutting the C-termiminus at the same positions: T183 (G. gallus), T182 (D. rerio) or S244 (C. intestinalis). Then ASAP1 variants and Archlight Q239 were cloned between the NheI-HindIII sites of pcDNA3.1/Puro-CAG. The scientists used coexpressed sensors with a
Get Access