We characterized an activation mechanism of the human LTRPC2 protein, a member of the transient receptor potential family of ion channels, and demonstrated that LTRPC2 mediates Ca2+ influx into immunocytes. Intracellular pyrimidine nucleotides, adenosine 5'-diphosphoribose (ADPR), and nicotinamide adenine dinucleotide (NAD), directly activated LTRPC2, which functioned as a Ca2+-permeable nonselective cation channel and enabled Ca2+ influx into cells. This activation was suppressed by intracellular adenosine triphosphate. These results reveal that ADPR and NAD act as intracellular messengers and may have an important role in Ca2+ influx by activating LTRPC2 in immunocytes.

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To find a novel human ion channel gene we have executed an extensive search by using a human genome draft sequencing data base. Here we report a novel twopore domain K ؉ channel, TRESK (TWIK-related spinal cord K ؉ channel). TRESK is coded by 385 amino acids and shows low homology (19%) with previously characterized two-pore domain K ؉ channels. However, the most similar channel is TREK-2 (two-pore domain K ؉ channel), and TRESK also has two pore-forming domains and four transmembrane domains that are evolutionarily conserved in the two-pore domain K ؉ channel family. Moreover, we confirmed that TRESK is expressed in the spinal cord. Electrophysiological analysis demonstrated that TRESK induced outward rectification and functioned as a background K ؉ channel. Pharmacological analysis showed TRESK to be inhibited by previously reported K ؉ channel inhibitors Ba 2؉ , propafenone, glyburide, lidocaine, quinine, quinidine, and triethanolamine. Functional analysis demonstrated TRESK to be inhibited by unsaturated free fatty acids such as arachidonic acid and docosahexaenoic acid. TRESK is also sensitive to extreme changes in extracellular and intracellular pH. These results indicate that TRESK is a novel two-pore domain K ؉ channel that may set the resting membrane potential of cells in the spinal cord.

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This prospective study was designed to evaluate whether static stretching can prevent training-related injuries in Japan Ground Self-Defense Force military recruits. A total of 901 recruits between 1996 and 1998 were divided into two groups. Of which, 518 recruits were assigned to the stretching group and practiced static stretching before and after each physical training session. The control subjects (383 recruits in the nonstretching group) did not stretch statically prior to exercise. The static stretching consisted of 18 exercises. We collected injury data from medical records and assessed the incidence and the location of injury. The total injury rate was almost the same between two groups; however, the incidences of muscle/tendon injury and low back pain were significantly lower in the stretching group (p < 0.05). Static stretching decreased the incidence of muscle-related injuries but did not prevent bone or joint injuries.

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The response to intracellular ADP-ribose in the rat CRI-G1 insulinoma cell line was studied using a patch-clamp method. Dialysis of ADP-ribose into cells induced a response in a dose-dependent manner. The reversal potentials in various solutions showed that the ADP-ribose-gated channel was a Ca2+-permeable nonselective cation channel. In inside-out recordings, ADP-ribose and b-NAD induced responses in the same patch. The single-channel current-voltage relationships for ADP-ribose- and b-NAD-induced responses were almost identical, indicating that ADP-ribose and b-NAD activated the same channel. The physiological properties of the ADP-ribose-gated channel are similar to those we reported previously for the cloned transient receptor potential channel TRPM2. Moreover, RT-PCR analysis showed that TRPM2 was abundantly expressed in CRI-G1 cells, suggesting that the ADP-ribose-gated channel represents the native TRPM2 channel in CRI-G1 cells. These results suggest that ADP-ribose can be an endogenous modulator of Ca2+ influx through the TRPM2 channel into CRI-G1 cells.

We report identification and characterization of Kv6.3, a novel member of the voltage-gated K + channel. Reverse transcriptase-polymerase chain reaction analysis indicated that Kv6.3 was highly expressed in the brain. Electrophysiological studies indicated that homomultimeric Kv6.3 did not yield a functional voltage-gated ion channel. When Kv6.3 and Kv2.1 were co-expressed, the heteromultimeric channels displayed the decreased rate of deactivation compared to the homomultimeric Kv2.1 channels. Immunoprecipitation studies indicated that Kv6.3 bound with Kv2.1 in co-transfected cells. These results indicate that Kv6.3 is a novel member of the voltage-gated K + channel which functions as a modulatory subunit. ß