Menthol stimulates calcium absorption in the rumen but not in the jejunum of sheep

Stimulation of Ca²⁺ absorption can counteract hypocalcemia at the onset of lactation. The plant bioactive lipid compound (PBLC) menthol is an agonist for nonselective cation channels of the transient receptor potential (TRP) family. It acutely stimulated Ca²⁺ absorption in ruminal epithelia of nonadapted animals ex vivo and caused higher plasma Ca²⁺ concentrations in cows and sheep in vivo. To elucidate the pathway by which menthol feeding increases plasma Ca²⁺ level, the present study aimed to investigate the long-term dose-dependent effects of dietary menthol-rich PBLC on Ca²⁺ absorption and mRNA abundances of TRP channels in both rumen and jejunum. Twenty-four growing Suffolk sheep were equally distributed to a Con, PBLC-L, and PBLC-H group, which received 0, 80, and 160 mg/d of a menthol-rich PBLC. After 4 wk, ruminal and jejunal epithelia were analyzed for mRNA abundances of TRPA1, TRPV3, TRPV5–6, and TRPM6–8 genes. The Ca²⁺ flux rates and electrophysiological properties of epithelia from rumen and mid-jejunum were measured in Ussing chambers in the presence and absence of mucosal Na⁺. Acute changes in Ca²⁺ flux rates were measured after mucosal application of 50 µM menthol. Ruminal epithelia had quantifiable transcripts of TRPV3 = TRPM6 > TRPM7 > TRPA1 with no difference among feeding groups. Jejunum had quantifiable transcripts of TRPM7 > TRPA1 ≥ TRPM6 ≥ TRPV6 > TRPV5, where TRPA1, TRPV5, and TRPV6 tended to decrease linearly with increasing PBLC dose. Absorptive net flux of Ca²⁺ was detected only in the rumen, whereas jejunum showed a high passive permeability to Ca²⁺. Net flux rates of Ca²⁺ in the rumen increased in a quadratic manner (highest in PBLC-L animals) and were systematically decreased with the omission of mucosal Na⁺. Short-circuit current increased in both PBLC feeding groups compared with Con only in the rumen. Acute application of menthol-stimulated mucosal-to-serosal and net Ca²⁺ flux rates only in ruminal epithelia with higher stimulation in PBLC-fed animals. We conclude that Ca²⁺ transport is mainly active and transcellular in the rumen. It most likely involves TRPV3 that can be stimulated by menthol. Pre-feeding of menthol-rich PBLC enhances ruminal Ca²⁺ absorption and sensitizes it to acute stimulation by menthol. By contrast, intestinal Ca²⁺ absorption is not sensitive to menthol stimulation. Menthol could be used as a tool to enhance ruminal Ca²⁺ absorption and to prevent hypocalcemia in dairy cows.