Differential proteome profiling in the hippocampus of amnesic mice

Abstract

Amnesia or memory loss is associated with brain aging and several neurodegenerative pathologies including Alzheimer's disease (AD). This can be induced by a cholinergic antagonist scopolamine but the underlying molecular mechanism is poorly understood. This study of proteome profiling in the hippocampus could provide conceptual insights into the molecular mechanisms involved in amnesia. To reveal this, mice were administered scopolamine to induce amnesia and memory impairment was validated by novel object recognition test. Using two-dimensional gel electrophoresis coupled with MALDI-MS/MS, we have analyzed the hippocampal proteome and identified 18 proteins which were differentially expressed. Out of these proteins, 11 were downregulated and 7 were upregulated in scopolamine-treated mice as compared to control. In silico analysis showed that the majority of identified proteins are involved in metabolism, catalytic activity, and cytoskeleton architectural functions. STRING interaction network analysis revealed that majority of identified proteins exhibit common association with Actg1 cytoskeleton and Vdac1 energy transporter protein. Furthermore, interaction map analysis showed that Fascin1 and Coronin 1b individually interact with Actg1 and regulate the actin filament dynamics. Vdac1 was significantly downregulated in amnesic mice and showed interaction with other proteins in interaction network. Therefore, we silenced Vdac1 in the hippocampus of normal young mice and found similar impairment in recognition memory of Vdac1 silenced and scopolamine-treated mice. Thus, these findings suggest that Vdac1-mediated disruption of energy metabolism and cytoskeleton architecture might be involved in scopolamine-induced amnesia.

Transcriptional co‐repressor SIN3A silencing rescues decline in memory consolidation during scopolamine‐induced amnesia

“…The SIN3A siRNA sequence (sense 5ˊ-CUACGUCUCAAGGAAC-CUtt-3ˊ, antisense 5 0 -UAGGUUCCUUGAGAC GUAGtt-3 0 ) was validated from NCBI (https://www.ncbi.nlm.nih.gov/nuccore/). Furthermore, 3 lg SIN3A and scrambled siRNA were infused in dorsal hippocampi bilaterally, as suggested in manufacturer's protocol and earlier reports (Singh et al 2015;Baghel and Thakur 2017). Initially, we have assessed the silencing efficiency of siRNA using administration of 3 lg SIN3A siRNA diluted in 0.9% NaCl in nuclease-free water, scrambled siRNA and control (administered with water) in each hippocampus.…”

Natural silibinin modulates amyloid precursor protein processing and amyloid-β protein clearance in APP/PS1 mice

“…Based on these studies, silibinin potentially acts on multiple targets to inhibit AD pathologies. However, the effects of In the present study, learning and memory deficits were assessed using the Y-maze test, novel object recognition test and Morris water maze test, which are widely used methods to investigate the behaviors of mice [17][18][19][20]. The contents of soluble and insoluble Aβ and Aβ plaques are increased in 6-month-old APP/PS1 mice, comparable to patients with AD [21,22].…”

Lymphatic filarial serum proteome profiling for identification and characterization of diagnostic biomarkers

“…After IEF, the strips were first equilibrated with 40 mM Tris-HCl buffer (pH 8.8) containing 6 M urea, 25% w/v glycerol, 2% w/v SDS, 1% w/v DTT and 2.5% iodoacetamide. The second dimension was done on 10% SDS PAGE followed by staining with Coomassie Brilliant Blue G-250 (10% Aluminum sulfate, 10% ethanol, 0.02% CBB G-250, and 2.5% ortho-phosphoric acid), and images were taken with a gel documentation system (Alpha Innotech, USA) and analyzed by PDQuest software (BioRad, USA) [ 23 ]. Three independent experiments were performed to ensure the reproducibility of results.…”