Mechanosensitive channel gating has been suggested to occur via two models: the bilayer model, whereby forces generated within the lipid bilayer serve to gate the mechanosensitive channel, and the tethered model, whereby cytoskeletal tension plays a role in mechanosensitive channel gating. With respect to stretch-inactivated mechanosensitive channels in supraoptic neurons that are important in osmoreception, it is unclear which model prevails. To determine if the tethered model, and therefore the cytoskeleton, plays a role in mechanosensitive channel gating, neurons were subjected to drugs that function to depolymerize (Cytochalasin D; CytD) or stabilize (Jasplakinolide; Jasp) the actin cytoskeleton. A mechanosensitivity index was calculated in control, CytD and Jasp conditions by performing whole cell voltage clamp recordings on acutely isolated supraoptic neurons (magnocellular neurosecretory cells; MNCs) from adult Long Evans rats. Conductance and volume data revealed that cells treated in Cytochalasin D possessed a decreased mechanosensitivity compared to control, whereas cells treated with Jasplakinolide possessed an increased mechanosensitivity. The results provide evidence that actin is involved in mechanosensitive channel gating in supraoptic neurons and suggest that mechanosensitive gating occurs via the tethered model.