Processing bodies (P-bodies) are subcellular ribonucleoprotein (RNP) granules that have been hypothesized to be sites of mRNA degradation, mRNA translational control, and/or mRNA storage. Importantly, P-bodies are conserved from yeast to mammals and contain a common set of evolutionarily conserved protein constituents. P-bodies are dynamic structures and their formation appears to fluctuate in correlation with alterations in mRNA metabolism. Despite these observations, little is understood about how P-body structures are formed within the cell. In this study, we demonstrate a relationship between P-bodies and microtubules in the budding yeast, Saccharomyces cerevisiae. First, we demonstrate that disruption of microtubules by treatment with the drug benomyl leads to aggregation of P-body components. Consistent with this finding, we also demonstrate that disruption of microtubules by a temperature-sensitive allele of the major a tubulin, TUB1 (tub1-724) stimulates P-body formation. Second, we find that the a-tubulin protein Tub1 colocalizes with P-bodies upon microtubule destabilization. Third, we determine that a putative tubulin tyrosine ligase, encoded by YBR094W, is a protein component of P-bodies, providing additional evidence for a physical connection between P-bodies and microtubules. Finally, we establish that P-bodies formed by microtubule destabilization fail to correlate with global changes in the stability of mRNA or in general mRNA translation. These findings demonstrate that the aggregation of P-body components is linked to the intracellular microtubule network, and, further, that P-bodies formed by disruption of microtubules aggregate independent of broad alterations in either mRNA decay or mRNA translation.

Sixty-four male Sprague-Dawley rats were randomly assigned to one of eight treatment groups to determine the effects of vitamin E (VitE), dehydroepiandrosterone (DHEA), and exercise on antioxidant status in plasma and skeletal muscle. Indexes of oxidative stress were determined by measuring two markers of lipid peroxidation and the activity of two free radical scavenging enzymes. One-half of the rats had their diets supplemented with 250 IU VitE/kg of diet. One-half of the rats were injected with 0.35 mol/kg body wt ip of DHEA-acetate, whereas the others were injected with vehicle. All treatments lasted 5 wk. Before being killed, one-half of each treatments group of rats was randomly assigned to run for 1 h on a motorized rodent treadmill at 21 m/min up a 12% grade. The other rats remained rested before being killed. Exercise increased thiobarbituric acid-reactive substances (TBARS) and lipid hydroperoxides in plasma and TBARS in red slow-twitch and white fast-twitch muscles. VitE reduced the amount of lipid hydroperoxides and TBARS in plasma and TBARS in all three muscle fiber types. VitE also reduced glutathione peroxidase (GPX) activity in plasma and red fast-twitch muscle. DHEA increased indexes of oxidative stress in plasma and white fast-twitch muscle. DHEA reduced GPX activity in plasma but increased GPX activity in all three muscle fiber types. These results indicate that aerobic exercise is a mild oxidative stressor with DHEA exacerbating this response and that VitE helps diminish this effect in certain muscle fiber types.

In infants, respiratory infection elicits tachypnea. To begin to evaluate the role of brainstem cytokine expression in modulation of breathing pattern changes, we compared the pattern generated after endotracheal instillation of lipopolysaccharide (LPS) in in vivo rat pups to local pro-inflammatory cytokine injection in the nucleus tractus solitarius (nTS) in an in vitro en bloc brainstem spinal cord preparation. We hypothesized that both challenges would elicit similar changes in patterning of respiration. In anesthetized, spontaneously breathing rat pups, lipopolysaccharide (LPS) or saline was instilled in the airway of urethane-anesthetized rats (postnatal day 10–11). We recorded diaphragm EMG over the subsequent 2 hours and saw a 20–30% decrease in interburst interval (Te) at 20–80 min post-injection in LPS-instilled animals with no significant change in Ti. In contrast, IL-1β injections into the nTS of en bloc in vitro brainstem-spinal cord preparations from 0 to 5 day-old pups maintained Ti and caused an increase in Te as early as 20 min later, decreasing frequency for 80 to 120 minutes after injection. Our results suggest that the neonatal respiratory response to the cytokine IL-1β mediated inflammatory response depends on the site of the inflammatory stimulus and that the direct effect of IL-1β in the nTS is to slow rather than increase rate.

Sixty-four male Sprague-Dawley rats were randomly assigned to one of eight treatment groups to determine whether vitamin E (VitE) could help protect the heart from oxidative stress induced by either dehydroepiandrosterone (DHEA) or exercise. Oxidative stress was indicated by lipid peroxidation [i.e., thiobarbituric acid-reactive substances (TBARS)] and two scavenger enzymes. VitE supplementation (250 IU VitE/kg of diet) was given to one-half of the rats. DHEA acetate (0.35 mol/kg body wt) was injected intraperitoneally to one-half of the animals while the others were injected with corn oil vehicle. All treatments lasted for 5 wk. Next, 32 rats were randomly assigned to run for 1 h on a motorized rodent treadmill at 21 m/min up a 12% grade and then were killed. The remaining rats were killed at rest. Exercise increased TBARS in heart independent of treatment (1.94 +/- 0.12 vs. 2.43 +/- 0.11 nmol/mg protein). VitE attenuated the amount of TBARS in heart when DHEA was given. DHEA significantly increased TBARS in heart. Total and selenium-dependent glutathione peroxidase activities in heart were unaffected by any treatment. DHEA increased catalase activity at rest. Exercise increased catalase activity (71.5 +/- 7.9 vs. 97.4 +/- 9.5 mu mol x min-1 x mg protein-1); however, when VitE was given, the response to exercise was attenuated (74.1 +/- 8.4 vs. 80.9 +/- 9.9 mu mol center dot min-1 x mg protein-1). These results suggest that aerobic exercise and DHEA are mild oxidative stressors on the heart and that VitE supplementation can be beneficial in attenuating these combined stressors on the heart.

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