Elastic properties of the cell surface and metabolic profile of an embryonic primary mixed culture of hippocampal neurons under conditions of P2X3 receptor blockade

P2X3-receptors localized in the hippocampus participate in the transmission of excitation and the formation of synaptic plasticity underlying learning and memory. P2X3-receptors are of great importance in the occurrence of neuropathic pain in epilepsy, acute and inflammatory pain of various genesis and localization as well as in the activation and growth of nerves after traumatic brain injury. The aim of the study was to study the elastic properties of the surface and the metabolic profile of neurons in an embryonic primary mixed hippocampal culture under P2X3-receptor blockade. The study was performed on a primary mixed culture of hippocampal neurons obtained from CD1 mice on the 18th day of gestation (E18). The highly selective blocker 5-(5-iodo-2-isopropyl-4-methoxyphenoxy)pyrimidine-2.4-diamine monochloride salt was selected as a P2X3-receptor blocker. To assess the elastic properties of neurons Young's modulus that characterizes the rigidity of the cell surface was measured. Measurements on an atomic force microscope applying a load in 25 local areas of the cell surface were performed. At each point, the force curves of the cantilever approach and retraction were recorded with subsequent calculation of Young's modulus. The metabolic profile of the neuroglial culture in Energy Phenotype test on a Seahorse HS mini cell metabolism analyzer (USA) was studied. The Young's modulus of the cell surface of neurons in the control was in the range from 6.8 ± 0.1 to 9.7 ± 0.2 kPa, and under the action of the P2X3-receptor blocker in the range from 3.1 ± 0.1 kPa to 8.5 ± 0.1 kPa. Under the conditions of P2X3-receptor blockade on the 5th day of differentiation the Young's modulus of the cell surface was reduced by 62% (p 0.05), on the 8th day it increased by 22% (p 0.05) and by the 11th day it decreased by 16.7% (p 0.05) compared to the control. Aerobic respiration was characteristic of the embryonic hippocampal culture both in the control and with the P2X3-receptor blockade. Consequently, the blockade of the P2X3-receptor did not affect the metabolic profile of the E18 hippocampal culture. The obtained data indicate the direct participation of the P2X3-receptor in the formation of biomechanical properties of the cell surface in the processes of differentiation and signal transduction. It is possible, that the blockade of the P2X3-receptor will be one of the promising molecular targets that can reduce neuronal damage in brain injuries, neuroinflammation, hypoxia, and epilepsy. In addition, the study of the P2X3-receptor blockade can expand the fundamental understanding of the role of the purinergic signaling system in the formation of complex neuronal morphology at early stages of embryonic development under conditions of rapid excitatory signal transmission mediated by the ATP molecule.