Journal: bioRxiv

Article Title: Characterization of β-Hydroxybutyrate as a Cell Autonomous Fuel for Active Excitatory and Inhibitory Neurons

doi: 10.1101/2024.06.08.598077

Figure Lengend Snippet: A) Synaptic vesicle recycling following 100 action potential (AP), 10 Hz stimulus in excitatory neurons expressing CaMKII synaptophysin-pHluorin (physin-pHluorin), while fueled by 5 mM glucose or 5 mM β-hydroxybutyrate (BHB) (n=9, error bands SEM). B) Synaptic vesicle cycling in inhibitory neurons expressing hDlxI56i physin-pHluorin and fueled by glucose or BHB during 100 AP, 10 Hz stimulus (n=4, error bands SEM). C) Schematic of ketone catabolism in neurons by β-hydroxybutyrate dehydrogenase 1 (BDH1), succinyl-CoA:3-ketoacid CoA transferase (SCOT), and thiolase II (T2). D) Representative Western blot against BDH1 in WT cortical culture or culture treated with BDH1 shRNA virus for 10 days (full blots in Extended Data Figure 1-1, -2, -3, -4, -5, -6, -7, -8). E) Quantification of BDH1 knockdown efficiency in Western blot, normalized to average signal of uninfected controls (N=4 cultures, n=4 technical replicates, mean=0.6238. Error bars SEM, *p=0.011, Wilcoxon matched-pairs signed rank test). F) Synaptic vesicle recycling following 100 AP, 10 Hz stimulus in excitatory neurons expressing physin-pHluorin and shRNA against BDH1, while fueled by 5 mM glucose or BHB (n=4, error bands SEM). G) Synaptic vesicle cycling in inhibitory neurons expressing physin-pHluorin with and without shRNA against BDH1, while fueled by glucose or BHB during 100 AP, 10 Hz stimulus (n=4, error bands SEM).

Article Snippet: For SCOT knockdown validation, OXCT1 Rabbit polyclonal antibody (Proteintech 12175-1-AP) was used at 1:2000.

Techniques: Expressing, Western Blot, shRNA, Virus, Knockdown