The bovine TRPV3 as a pathway for the uptake of Na(+), Ca(2+), and NH(4)(+)

Absorption of ammonia from the gastrointestinal tract results in problems that range from hepatic encephalopathy in humans to poor nitrogen efficiency of cattle with consequences for the global climate. Previous studies on epithelia and cells from the native ruminal epithelium suggest functional involvement of the bovine homologue of TRPV3 (bTRPV3) in ruminal NH(4)(+) transport. Since the conductance of TRP channels to NH(4)(+) has never been studied, bTRPV3 was overexpressed in HEK-293 cells and investigated using the patch-clamp technique and intracellular calcium imaging. Control cells contained the empty construct. Divalent cations blocked the conductance for monovalent cations in both cell types, with effects higher in cells expressing bTRPV3. In bTRPV3 cells, but not in controls, menthol, thymol, carvacrol, or 2-APB stimulated whole cell currents mediated by Na(+), Cs(+), NH(4)(+)(,) and K(+), with a rise in intracellular Ca(2+) observed in response to menthol. While only 25% of control patches showed single-channel events (with a conductance of 40.8 ± 11.9 pS for NH(4)(+) and 25.0 ± 5.8 pS for Na(+)), 90% of bTRPV3 patches showed much larger conductances of 127.8 ± 4.2 pS for Na(+), 240.1 ± 3.6 pS for NH(4)(+), 34.0 ± 1.7 pS for Ca(2+), and ~ 36 pS for NMDG(+). Open probability, but not conductance, rose with time after patch excision. In conjunction with previous research, we suggest that bTRPV3 channels may play a role in the transport of Na(+), K(+), Ca(2+) and NH(4)(+) across the rumen with possible repercussions for understanding the function of TRPV3 in other epithelia.