Magnesium-l-threonate (L-TAMS) attenuates the mechanical allodynia and thermal hyperalgesia and prevents magnesium deficiency induced by vincristine (VCR). The time courses of paw withdrawal threshold (A) and paw withdrawal latency (B) in vehicle (Veh), L-TAMS, VCR, and L-TAMS + VCR groups are shown. Treatment with L-TAMS significantly suppressed VCR-induced mechanical allodynia and thermal hyperalgesia (n = 12 per group). (C, D) Free Mg²⁺ concentrations in serum and in cerebrospinal fluid of different groups measured with magnesium green (MG) at different time point are shown. The number of animals used in each group is indicated in the histograms. (E) Representative flow physical diagram and histograms show the level of intracellular free magnesium of dorsal root ganglion (DRG) neurons (assessed as mean fluorescence intensity [MFI] of 30,000 cells per rat). The number of animals used in each group is indicated in the histograms. *P < 0.05, **P < 0.01, ***P < 0.001, compared with the Veh group; #P < 0.05, ##P < 0.01, compared with the VCR group. FITC-H = fluorescein isothiocyanare-high; FS = forward scatter; SS = side scatter.