Ciliobrevin D

Ciliobrevin D (compound 5) is a cell-permeable, reversible and specific antagonist of AAA+ (ATPases associated with diverse cellular activities) ATPase motor cytoplasmic dynein. Ciliobrevin D perturbs primary cilia formation and blocks Hedgehog (Hh) signaling.

Ciliobrevin D inactivate dynein in Sertoli cells. The inactivation of dynein by ciliobrevin D also perturbs gross disruption of F-actin across the Sertoli cells in vitro.^[2] Ciliobrevin D perturbs protein trafficking within the primary cilium, leading to their malformation and Hedgehog signaling blockade. Ciliobrevin D also prevents spindle pole focusing, kinetochore-microtubule attachment, melanosome aggregation, and peroxisome motility in cultured cells.^[1]

Sertoli cells cultured in vitro on Matrigel-coated bicameral units at 1.0 × 10⁶ cells/cm² are used for quantifying the transepithelial electrical resistance in ohms (Ω) across the cell epithelium to assess the TJ-barrier function. In brief, each bicameral unit is placed inside the well of a 24-well dish with 0.5 ml F12/DMEM in the apical and the basal compartments. Dynein is inhibited in Sertoli cells on days 3 and 4 using Ciliobrevin D at 15 µM or 30 µM for 1hr, and Sertoli cell TJ-permeability barrier function is monitored daily by quantifying transepithelial electrical resistance across the cell epithelium.

Ciliobrevin D inactivate dynein in the testis. The inactivation of dynein by ciliobrevin D also perturbs gross disruption of F-actin across the seminiferous epithelium in vivo, illustrating there are cross talks between the two cytoskeletons in the testis.^[2]