Journal: Cureus

Article Title: Role of Adenosine A1 Receptor in Sleep Deprivation-Induced Neuroinflammation: Insights on Rapid Eye Movement Sleep and Fear Extinction Memory Recall in Rats

doi: 10.7759/cureus.75926

Figure Lengend Snippet: The study involved the following experimental timeline: 1) habituation and baseline behavioural recordings, 2) fear conditioning, 3) fear extinction training, 4) animals divided into three groups (a. CC (ad libitum sleep), b. SD (sleep deprivation for 48 hours + vehicle, i.e., 20% DMSO), c. SD + CPT (8-cyclopentyltheophylline, adenosine A1 receptor antagonist was administered for two days during 48-hour SD exposure), 5) fear extinction recall test and depressive behaviour assessment, 6) sleep architecture recording, 6) brain region dissection for molecular analyses. Out of the 44 animals screened for locomotor activity, 39 animals qualified for behavioural experiments. Five animals were excluded due to higher anxiety levels and abnormal locomotor activity. All experimental animals were handled and habituated by the experimenter, twice a day, for three days. Then, the animals were subjected to cued fear conditioning on day 1. After 24 hours, cued fear extinction training was given on day 2. Then, the animals were divided into three groups: CC (ad libitum sleep), SD (sleep deprivation for 48 hours + vehicle, i.e., 20% DMSO), SD + CPT (8-cyclopentyltheophylline, adenosine A1 receptor antagonist was administered for two days during the 48-hour SD exposure). Immediately, after the SD 48-hour exposure, all the experimental animals including control animals with ad libitum sleep were tested for the cued fear extinction recall test, open field test (OFT) on day 4, sucrose preference test for eight hours, forced swim test (FST) on day 5, followed by the dissection and collection of the brain sample. Brain samples were processed further for Immunohistochemistry (IHC), RT-PCR, ELISA, and high-performance liquid chromatography (HPLC) experiments. ‘n’ refers to sample size/number of animals in each group.

Article Snippet: Assessment of Fear Extinction Memory Recall Apparatus: The fear conditioning experiment was carried out in a chamber (Context A) constructed with Plexiglass cages and aluminium walls (Ugo Basile instruments, Italy) on day 1.

Techniques: Dissection, Activity Assay, Control, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, High Performance Liquid Chromatography

Journal: Cureus

Article Title: Role of Adenosine A1 Receptor in Sleep Deprivation-Induced Neuroinflammation: Insights on Rapid Eye Movement Sleep and Fear Extinction Memory Recall in Rats

doi: 10.7759/cureus.75926

Figure Lengend Snippet: A. Schematic representation of fear conditioning and cued extinction recall paradigm. B. Statistical comparison graphs (two-way ANOVA, Tukey’s post-hoc, n = 11) depicting the freezing scores exhibited during fear conditioning (day 1), fear extinction (day 2), and fear extinction recall (day 4) post either 48-hour sleep deprivation with vehicle (DMSO) (F(4, 60) = 15.8; p < 0.001; n = 11) or A1R antagonist CPT (p = 0.01; n = 11), and ad libitum sleep (cage control group). *p < 0.05 represents the comparison between CC and SD; #p < 0.05 represents the comparison between SD and SD + CPT. CC: cage control; SD: 48-hour sleep deprivation; SD + CPT: 48-hour sleep deprivation + 8-cyclopentyltheophylline

Article Snippet: Assessment of Fear Extinction Memory Recall Apparatus: The fear conditioning experiment was carried out in a chamber (Context A) constructed with Plexiglass cages and aluminium walls (Ugo Basile instruments, Italy) on day 1.

Techniques: Comparison, Control

Journal: Nature Communications

Article Title: A pH-sensitive closed-loop nanomachine to control hyperexcitability at the single neuron level

doi: 10.1038/s41467-024-49941-3

Figure Lengend Snippet: A Schematics of the bilateral stereotaxic injection of AAV2/1, encoding either Ctrl or pHIL under the CaMKIIα promoter, in the dorsal hippocampus of 2-month-old wild type C57BL/6 mice followed, one month later, by whole-body bioluminescence imaging and behavioral tests after vehicle or CTZ 400a (CTZ) administration (Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license ( https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en )). B Left: Representative whole-body bioluminescence images (merged emission of both RLuc8 and E 2 GFP) of Ctrl ( top row ) and pHIL ( bottom row ) transduced mice acquired every 5 min after the intravenous injection of CTZ (0.3 mg/kg). Radiance intensity is shown in pseudocolors. Right: The quantitative evaluation of the mean (± SEM) RLuc8/E 2 GFP live average radiance values (means ± SEM) in Ctrl ( black bars ) and pHIL ( red bars ) transduced mice displays an early emission peak 5 min after CTZ administration, followed by a progressive decrease ( n = 11 mice for both Ctrl and pHIL). C Left: Representative whole-body bioluminescence images acquired 5 min after the administration of increasing doses (0.15, 0.3 and 0.6 mg/kg) of CTZ to pHIL-transduced mice. Right: Corresponding RLuc8/E 2 GFP live average radiance values (means ± SEM) for the three doses as a function of time after CTZ administration. For further details see panel B ( n = 6, 10, 5 mice for 0.15, 0.3 and 0.6 mg/kg). D – F The locomotor activity and hippocampus-dependent behavior were investigated in pHIL-transduced mice upon administration of either CTZ (0.3 and 0.6 mg/kg) or the respective vehicle. The control condition (dose = 0 mg/kg) refers to untreated pHIL-transduced mice. D Open field test. The total distance covered by the mice ( top ) and the time spent in the center or along the border ( bottom ) were comparable under all tested conditions, proving no interference of the pharmaceutical treatment with locomotor activity (means ± SEM of n = 7, 7, 10 for 0, 0.3 and 0.6 mg/kg respectively in both Veh and CTZ groups). E Novel object recognition. No effects of CTZ were observed both in the familiarization phase ( top ) and in the recognition phase ( bottom ) when mice were exposed to one object previously explored and a novel unfamiliar object (means ± SEM of n = 7, 7, 7 for 0, 0.3 and 0.6 mg/kg respectively in both Veh and CTZ groups). F Contextual fear conditioning. No significant differences in freezing times were observed both during the fear conditioning phase ( top ) and in control exposure to a new context ( bottom ) (means ± SEM of n = 7, 7, 7 for 0, 0.3, and 0.6 mg/kg respectively in both Veh and CTZ groups). In E and F, either CTZ or vehicle was administered before the novel recognition phase and the conditioning session, respectively. p > 0.05, two-way repeated measures ANOVA (D-F).

Article Snippet: The contextual fear-conditioning test was conducted in the Video Fear Conditioning apparatus (Med Associates Inc., Fairfax, VT).

Techniques: Injection, Imaging, Activity Assay, Control