Expcsure of adult male albino rats to higher environmental temperature (HET) at 35° for 2-12 hr or at 45° for 1-2 hr increases hypothalamic synaptosomal acetylcholinesterase (AChE) activity. Synaptosomal AChE activity in cerebral cortex of rats exposed to 35° for 12 hr and in cerebral cortex and pons-medulla of rats exposed to 45° for 1-2 hr are also activated. AChE activity of synaptosomes prepared from normal rat brain regions incubated in-vitro at 39° or 41° for 0.5 hr increases significantly in cerebral cortex and hypothalamus. The activation of AChE in ponsmedulla is also observed when this brain region is incubated at 41° for 0.5 hr. Increase of (a) the duration of incubation at 41° and (b) the incubation temperature to 43° under in-vitro condition decreases the synaptosomal AChE activity. Lioneweaver-Burk plots indicate that (a) in-vivo and invitro HET-induced increases of brain regional synaptosomal AChE activity are coupled with an increase of Vmax without any change in Km (b) very high temperature (43° under in-vitro condition) causes a decrease in Vmax with an increase in Km of AChE activity irrespective of brain regions. Arrhenius plots show that there is a decrease in transition temperature in hypothalamus of rats exposed to either 35° or 45°; whereas such a decrease in transition temperature of the pons-medulla and cerebral cortex regions are observed only after exposure to 45°. These results suggests that heat exposure increases the lipid fluidity of synaptosomal membrane depending on the brain region which may expose the catalytic site of the enzyme (AChE) and hence activate the synaptosomal membrane bound AChE activity in brain regions. Further the in-vitro higher temperature (43°C)-induced inhibition of synaptosomal AChE activity irrespective of brain regions may be the cause iof partial proteolysis/disaggregation of AChE oligomers and/or solubilization of this membrane-bound enzyme. © 1990 Plenum Publishing Corporation.