Dynasore

Dynasore (Dynamin Inhibitor I) is a highly selective, reversible small-molecule dynamin inhibitor with an IC₅₀ of 15 μM. Dynasore also inhibits the mitochondrial dynamin Drp1 without affecting other small GTPases. By inhibiting the GTPase activity of dynamin, Dynasore blocks its function in membrane fission, thereby inhibiting clathrin-mediated endocytosis. Dynasore can be used in research on endocytosis, membrane transport, viral entry, and receptor signaling regulation.

Dynamin1/2:15 μM, moderate potenc:~15 μM (vitro)

Methods: Mouse bone marrow-derived macrophages were pretreated with 5, 10, 25, or 50 μM Dynasore for 1 hour, then stimulated with 100 ng/mL LPS for 24 hours. Supernatants and cell lysates were collected. Pro-inflammatory cytokine expression levels were detected by real-time quantitative PCR.
Results: Dynasore dose-dependently inhibited LPS-induced mRNA and protein levels of proinflammatory cytokines (TNF-α, IL-6, IL-12). [1]
Methods: HCLE cells (Fluo-4 loaded) were pretreated with 40 μM Dynasore or 1 μM YM-58483 for 30 min, followed by induction of oxidative stress with 1 mM tBHP. Cytoplasmic calcium changes were monitored in real-time via fluorescence intensity over 2 hours.
Results: Dynasore completely blocked tBHP-induced increases in cytoplasmic calcium. [2]
Methods: Horizontal brainstem slices containing the solitary tract and solitary tract nucleus were prepared from adult rats. Whole-cell patch-clamp recordings were performed on NTS neurons at a clamp voltage of -60 mV. 100 μM Dynasore dissolved in artificial cerebrospinal fluid was continuously administered via a perfusion system. Postsynaptic currents were recorded at different time points (pre-administration, during administration, post-washout).
Results: Dynasore rapidly increased the frequency of spontaneous excitatory postsynaptic currents (EPSCs). Concurrently, ST-evoked EPSCs progressively failed until completely blocked, with no reversal during the recovery period.[3]

Methods: An acute lung injury (ALI) model was established in C57BL/6J mice via intratracheal LPS (10 mg/kg) administration. Dynasore (10, 30, 50 mg/kg) was administered as a single intraperitoneal injection 3 hours prior to LPS challenge. Mice were euthanized 24 hours after LPS stimulation.
Results: Dynasore pretreatment (particularly at 30 and 50 mg/kg) significantly reduced lung tissue histopathology scores and decreased inflammatory mediators. [1]

RIP1 kinase assay: Phosphorylation of RIP1 requires its kinase activity. Expression constructs of FLAGtagged wild-type (WT) or a kinase-inactive point mutant of RIP1 (K45M) are are transfected into 293T cells and RIP1 kinase assay is performed as described in the Methods in the presence of [γ-32P]ATP for 30 min at 30℃. Samples are subjected to SDS-PAGE and RIP1 band is visualized by autoradiography. Relative intensities of radioactive bands are quantified and are shown (ratio) in this and all other autoradiographs. In parallel to kinase reactions, a sample of beads is subjected to western blot analysis using anti-RIP1 antibody to ensure equal protein amounts in kinase reactions.

Mouse ventricular myocytes are isolated from male adult C6/Black mouse. Cardiomyocytes subjects to 2 hours of drug treatment followed by oxidative stress (30 μM Water2 for 35 min). For ATP supplement experiments, the cells are treated with 3 mM ATP for 30 min before exposure to Water2. Cardiomyocyte survival and viability are analyzed by trypan blue exclusion (TBE) assay[3].

Ethanol: 1.6 mg/mL (4.96 mM), Sonication is recommended.

DMSO: 120 mg/mL (372.31 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 4 mg/mL (12.41 mM), Sonication is recommended.