Citrus fruit and fabacea secondary metabolites potently and selectively block TRPM3.

BACKGROUND AND PURPOSE: The melastatin-related transient receptor potential TRPM3 is a calcium-permeable nonselective cation channel that can be activated by the neurosteroid pregnenolone sulphate (PregS) and heat. TRPM3-deficient mice show an impaired perception of noxious heat. Hence, drugs inhibiting TRPM3 possibly get in focus of analgesic therapy. EXPERIMENTAL APPROACH: Fluorometric methods were used to identify novel TRPM3-blocking compounds and to characterize their potency and selectivity to block TRPM3 but not other sensory TRP channels. Biophysical properties of the block were assessed using electrophysiological methods. Single cell calcium measurements confirmed the block of endogenously expressed TRPM3 channels in rat and mouse dorsal root ganglion (DRG) neurones. KEY RESULTS: By screening a compound library, we identified three natural compounds as potent blockers of TRPM3. Naringenin and hesperetin belong to the citrus fruit flavanones, and ononetin is a deoxybenzoin. Eriodictyol, a metabolite of naringenin and hesperetin, was still biologically active as a TRPM3 blocker. The compounds exhibited a marked specificity for recombinant TRPM3 and blocked PregS-induced [Ca(2+)]i signals in freshly isolated DRG neurones. CONCLUSION AND IMPLICATIONS: The data indicate that citrus fruit flavonoids are potent and selective blockers of TRPM3. Their potencies ranged from upper nanomolar to lower micromolar concentrations. Since physiological functions of TRPM3 channels are still poorly defined, the development and validation of potent and selective blockers is expected to contribute to clarifying the role of TRPM3 in vivo. Considering the involvement of TRPM3 in nociception, TRPM3 blockers may represent a novel concept for analgesic treatment.

Determination of organotin compounds in the foodweb of a shallow freshwater lake in the Netherlands.

An extensive study on the presence of nine organotin compounds (OTs) in a freshwater foodweb was made, using newly developed analytical procedures in order to obtain insight in accumulation and degradation processes. Tributyltin (TBT), Triphenyltin (TPT) and their degradation products were detected. Zebra mussels, eel, roach, bream, pike, perch, and pike perch and cormorant showed high OT body concentrations. At the lower trophic levels, phenyltin concentrations were higher in benthic species while butyltin concentrations were higher in pelagic species. This indicates that TBT is passed on primarily via the water, while TPT is passed on to a larger extent via the sediment. At the higher trophic levels, net bioaccumulation of TPT was greater than that of TBT, resulting in relatively higher TPT concentrations. High concentrations of biodegradation products of TBT, but not of TPT, were found in the livers of fish and birds, which indicates that TBT is more easily metabolized than TPT. A comparison with literature data of fish lethal body concentrations revealed that fish in the field may be endangered. With birds, the highest concentrations of OTs were present in liver and kidney and not in subcutaneous fat, which confirms that OTs accumulate via different mechanisms than traditional lipophilic compounds. As a whole the OT concentrations found in the foodweb may be considered to be quite alarming.