The upper part of a nodulated soybean root hydroponically cultured in a glass bottle was monitored using a computer microscope under controlled environmental conditions, and the diameter of individual nodules was measured from 10-24 d after planting. The diameter of a root nodule attached to the primary root increased from 1 mm to 6 mm for 2 weeks under nitrogen-free conditions. The increase in diameter of the nodules was almost completely stopped after 1 d of supplying 5 mM nitrate, and was due to the cessation of nodule cell expansion. However, nodule growth quickly returned to the normal growth rate following withdrawal of nitrate from the solution. The reversible depression of nodule growth by nitrate was similar to the restriction of photoassimilate supply by continuous dark treatment for 2 d followed by normal light/dark conditions. In addition, the inhibitory effect of nitrate was partially alleviated by the addition of 3% (w/v) sucrose to the medium. Plant leaves were exposed to (11)C or (14)C-labelled carbon dioxide to investigate the effects of 5 mM nitrate on the translocation and distribution of photosynthates to nodules and roots. Supplying 5 mM nitrate stimulated the translocation rate and the distribution of labelled C in nitrate-fed parts of the roots. However, the (14)C partitioning to nodules decreased from 9% to 4% of total (14)C under conditions of 5 mM nitrate supply. These results indicate that the decrease in photoassimilate supply to nodules may be involved in the quick and reversible nitrate inhibition of soybean nodule growth.

The ammonium ion is an indispensable nitrogen source for crops, especially paddy rice (Oryza sativa L. cv Nipponbare). Until now, it has been impossible to measure ammonium uptake and nitrogen movement in plants in real time. Using the new technologies of PETIS (positron emitting tracer imaging system) and PMPS (positron multi-probe system), we were able to visualize the real time translocation of nitrogen and water in rice plants. We used positron-emitting 13 N-labeled ammonium (13 NH 4 ϩ) and 15 O-water to monitor the movement. In plants cultured under normal conditions, 13 NH 4 ϩ supplied to roots was taken up, and a 13 N signal was detected at the discrimination center, the basal part of the shoots, within 2 minutes. This rapid translocation of 13 N was almost completely inhibited by a glutamine synthetase inhibitor, methionine sulfoximine. In general, nitrogen deficiency enhanced 13 N translocation to the discrimination center. In the dark, 13 N translocation to the discrimination center was suppressed to 40% of control levels, whereas 15 O-water flow from the root to the discrimination center stopped completely in the dark. In abscisic acid-treated rice, 13 N translocation to the discrimination center was doubled, whereas translocation to leaves decreased to 40% of control levels. Pretreatment with NO 3 Ϫ for 36 hours increased 13 N translocation from the roots to the discrimination center to 5 times of control levels. These results suggest that ammonium assimilation (from the roots to the discrimination center) depends passively on water flow, but actively on NH 4 ϩ-transporter(s) or glutamine synthetase(s).

The recently discovered family of RGS (regulators of G protein signaling) proteins acts as GTPase activating proteins which bind to ␣ subunits of heterotrimeric G proteins. We previously showed that a brain-specific RGS, RGS8 speeds up the activation and deactivation kinetics of the G protein-coupled inward rectifier K ؉ channel (GIRK) upon receptor stimulation (Saitoh, O., Kubo, Y., Miyatani, Y., Asano, T., and Nakata, H. (1997) Nature 390, 525-529). Here we report the isolation of a full-length rat cDNA of another brain-specific RGS, RGS7. In situ hybridization study revealed that RGS7 mRNA is predominantly expressed in Golgi cells within granule cell layer of cerebellar cortex. We observed that RGS7 recombinant protein binds preferentially to G␣ o , G␣ i3 , and G␣ z . When co-expressed with GIRK1/2 in Xenopus oocytes, RGS7 and RGS8 differentially accelerate G protein-mediated modulation of GIRK. RGS7 clearly accelerated activation of GIRK current similarly with RGS8 but the acceleration effect of deactivation was significantly weaker than that of RGS8. These acceleration properties of RGS proteins may play important roles in the rapid regulation of neuronal excitability and the cellular responses to short-lived stimulations.Numerous extracellular signals such as hormones, neurotransmitters, and odors stimulate seven transmembrane-spanning receptors that activate heterotrimeric G proteins. These G proteins function as signal transducing molecules by regulating cellular effectors including enzymes and ion channels (1, 2). The regulatory mechanisms that control G protein signaling have not been fully studied. Recently, a new family of regulators of G protein signaling (RGS) 1 was identified in organisms ranging from yeast to mammals (3, 4). Genetic screenings for negative regulators for pheromone response pathway in yeast identified a protein, Sst2 (5). By genetic and biochemical analyses, Sst2 was revealed to interact directly with G protein ␣ subunit (6). In the last few years, full or partial sequences of 22 RGS proteins have been identified in mammals. All of them share a conserved RGS domain of ϳ120 amino acids (7-15). It has been shown that several RGS proteins (RGS1, RGS3, RGS4, GAIP) attenuate G protein signaling in cultures (9,16,17). Biochemical studies have demonstrated that some RGS members (RGS1, RGS4, RGS10, GAIP, RGSr/RGS16, RET-RGS1) function as GTPase-activating proteins (GAPs) for the Gi family of ␣ subunit, including G␣ o , G␣ i , and transducin (10, 14, 18 -20). Hence, these characterized RGS proteins are proposed to down-regulate G protein signaling in vivo by enhancing the rate of G␣ GTP hydrolysis. However, whether other RGS proteins regulate G protein signalings in a similar manner remains to be established.Because there are many G protein signaling pathways which regulate important functions such as neural transmission in the brain, it is possible that certain RGS proteins might determine a mode of G protein signaling that control neural functions. We searched RGS proteins specifically ...

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