Methylcobalamin amplifies melatonin-induced circadian phase shifts by facilitation of melatonin synthesis in the rat pineal gland

Effects of methylcobalamin (methyl-B₁₂), a putative drug for treating human circadian rhythm disorders, on the melatonin-induced circadian phase shifts were examined in the rat. An intraperitoneal injection of 1–100 μg/kg melatonin 2-h before the activity onset time (CT 10) induced phase advances of free-running activity rhythms in a dose-dependent manner (ED₅₀=1.3 μg/kg). Injection of methyl-B₁₂ (500 μg/kg) prior to melatonin (1 μg/kg) injection induced larger phase advances than saline preinjected controls, while the injection of methyl-B₁₂ in combination with saline did not induce a phase advance. These results indicate amplification of melatonin-induced phase advances by methyl-B₁₂. Pinealectomy abolished the phase alternating effect of methyl-B₁₂, suggesting a site of action within the pineal gland. In fact, methyl-B₁₂ significantly increased the content of melatonin in the pineal collected 2-h after activity onset (CT 14). In contrast, no difference in melatonin content was found at CT 10, indicating that the effect of methyl-B₁₂ may be gated after the activity onset time when endogenous melatonin synthesis is known to increase. These results suggest that methyl-B₁₂ amplifies melatonin-induced phase advances via an increase in melatonin synthesis during the early subjective night at a point downstream from the clock regulation.