This work was carried out to study the antioxidant circadian system of two species of crayfish of different latitude origin. We investigated (1) whether both species possess glutathione circadian rhythms and (2) whether both species' rhythms differ in their ability to synchronize to 24 h cycles. Two batches of Procambarus clarkii and P. digueti were kept in (1) light–dark (LD) 12:12 low irradiance (LI) cycles and then exposed to (2) 72 h of complete darkness, (3) LD 12:12 high irradiance (HI), (4) LD 20:4 LI and (5) LD 20:4 HI for 2 weeks. The midgut and hemolymph were sampled and reduced and oxidized glutathione as well as glutathione reductase and glutathione peroxidase were assayed. Cosinor and analysis of variance revealed differences between both species. Procambarus clarkii robust antioxidant circadian rhythms are able to entrain to all conditions resetting to lights on or off. However, the P. digueti weak circadian glutathione system did not entrain to the LD cycles, showing a random distribution of phases. In this species, LD 12:12 and 20:4 HI evidenced significant daily rhythms indicating a damped circadian antioxidative system that is enhanced by the effect of light. This suggests that each species' photoperiodic history determines the adaptive abilities of the circadian antioxidative mechanisms.
The present study investigated the rhythmic changes in glutathione status in midgut gland and hemolymph as well as in glutathione reductase (GR) activity in the crayfish Procambarus clarkii. In order to determine the circadian nature of these rhythms different groups of crayfish were submitted to constant‐darkness conditions for 24 or 72 h after they had spent 15 days under light–dark 12:12 cycles. The animals of the different batches were killed at 6 h intervals during a 24 h cycle. Reduced glutathione (GSH) and oxidized glutathione (GSSG) in hemolymph and midgut as well as midgut GR activity were determined in midgut gland and hemolymph by fluorometric and spectrophotometric methods. Data analysis by chronogram and single Cosinor revealed circadian rhythmicity for GSH and GSSG concentration in both tissues as well as midgut GR activity. The rhythm parameters revealed oxidative stress induced by light. The possible correlation between the glutathione rhythm and other metabolic and behavioral rhythms of crayfish as well as the importance of the glutathione circadian temporal order in the adaptation of crayfish are discussed.
This work was carried out to study the antioxidant circadian system of two species of crayfish of different latitude origin. We investigated (1) whether both species possess glutathione circadian rhythms and (2) whether both species' rhythms differ in their ability to synchronize to 24 h cycles. Two batches of Procambarus clarkii and P. digueti were kept in (1) light-dark (LD) 12:12 low irradiance (LI) cycles and then exposed to (2) 72 h of complete darkness, (3) LD 12:12 high irradiance (HI), (4) LD 20:4 LI and (5) LD 20:4 HI for 2 weeks. The midgut and hemolymph were sampled and reduced and oxidized glutathione as well as glutathione reductase and glutathione peroxidase were assayed. Cosinor and analysis of variance revealed differences between both species. Procambarus clarkii robust antioxidant circadian rhythms are able to entrain to all conditions resetting to lights on or off. However, the P. digueti weak circadian glutathione system did not entrain to the LD cycles, showing a random distribution of phases. In this species, LD 12:12 and 20:4 HI evidenced significant daily rhythms indicating a damped circadian antioxidative system that is enhanced by the effect of light. This suggests that each species' photoperiodic history determines the adaptive abilities of the circadian antioxidative mechanisms.
The present study investigated the rhythmic changes in glutathione status in midgut gland and hemolymph as well as in glutathione reductase (GR) activity in the crayfish Procambarus clarkii. In order to determine the circadian nature of these rhythms different groups of crayfish were submitted to constant-darkness conditions for 24 or 72 h after they had spent 15 days under light-dark 12:12 cycles. The animals of the different batches were killed at 6 h intervals during a 24 h cycle. Reduced glutathione (GSH) and oxidized glutathione (GSSG) in hemolymph and midgut as well as midgut GR activity were determined in midgut gland and hemolymph by fluorometric and spectrophotometric method. Data analysis by chronogram and single Cosinor revealed circadian rhythmicity for GSH and GSSG concentration in both tissues as well as midgut GR activity. The rhythm parameters revealed oxidative stress induced by light. The possible correlation between the glutathione rhythm and other metabolic and behavioral rhythms of crayfish as well as the importance of the glutathione circadian temporal order in the adaptation of crayfish are discussed.
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