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Increased membrane unsaturation has been associated with shorter longevity due to higher sensitivity to lipid peroxidation (LP) leading to enhanced mitochondrial dysfunction and ROS overproduction. However, the role of LP during aging has been put in doubt along with the participation of electron leak at the electron transport chain (ETC) in ROS generation in aged organisms. Thus, to test these hypothesis and gain further information about how minimizing LP preserves ETC function during aging, we studied the effects of α-linolenic acid (C18:3) on in situ mitochondrial ETC function, ROS production and viability of chronologically aged cells of S. cerevisiae, whose membranes are intrinsically resistant to LP due to the lack of PUFA. Increased sensitivity to LP was observed in cells cultured with C18:3 at 6 days of aging. This was associated with higher viability loss, dissipated membrane potential, impaired respiration and increased ROS generation, being these effects more evident at 28 days. However, at this point, lower sensitivity to LP was observed without changes in the membrane content of C18:3, suggesting the activation of a mechanism counteracting LP. The cells without C18:3 display better viability and mitochondrial functionality with lower ROS generation even at 28 days of aging and this was attributed to full preservation of complex III activity. These results indicate that the presence of PUFA in membranes enhances ETC dysfunction and electron leak and suggest that complex III is crucial to preserve membrane potential and to maintain a low rate of ROS production during aging.

We evaluated different strains of the rotifer Asplanchna brightwellii collected from central Mexico using morphology, morphometry and molecular tools (CO1 and ITS). Three distinct clonal populations from each of the 3 regions (Mexico City, State of Mexico and State of Guerrero) were established under laboratory conditions. For a given waterbody, morphometric comparisons within the populations of A. brightwellii showed almost stable measurements of trophi and with no statistically significant differences among them (p > 0.05). However, asplanchnid body length and width as well as the cyst diameter varied significantly depending on the waterbody from which A. brightwellii was collected. The smallest adults (about 700 microm) were from Valerio Trujano lake (Guerrero State) samples while the largest were from Xochimilco lake. Similar tendencies were reflected in the diameter of resting eggs. In addition, morphologically the cysts of A. brightwellii from the three waterbodies showed slightly different pattern. The number of globular structures on the surface of cysts was smaller for Valerio Trujano strain, while these were larger and less numerous for both Xochimilco and Zumpango strains. The ITS region tree displayed two groups Xochimilco and Valerio Trujano -Zumpango, this analysis did not reflect the morphological grouping; on the contrary the CO1 gene tree separated the populations according to morphological clusters and location (Xochimilco, Valerio Trujano and Zumpango lakes). When the tree was built using the combination of both ITS and CO1 sequences, the phylogenetic relationships observed on CO1 gene were consistent; but showed differences with the relationships observed on ITS region tree (only two groups).

It has been previously reported that growth of Debaryomyces hansenii in 2 M NaCl induced the expression of ARO4. This gene codifies for DhAro4p, involved in the synthesis of the amino acid tyrosine. In this work we studied the activity of DhAro4p upon salt stress; a higher activity was observed in cells grown with 2 M NaCl, but tyrosine levels were not increased. On the other hand, the addition of tyrosine to the saline medium significantly enhanced the growth of D. hansenii. It was found that the oxidized form of tyrosine, 3-nitrotyrosine, increased in the presence of salt. Since NaCl protects against oxidative stress in D. hansenii (Navarrete et al., 2009), we propose that a protective pathway is the de novo synthesis of tyrosine and its immediate oxidation to 3-nitrotyrosine to counteract oxidative stress generated by salt stress, so we measured the production of reactive oxygen species (ROS) and nitric oxide (NO⁻) in D. hansenii after growing in 2 M NaCl. Results showed the presence of NO⁻ and the increased production of ROS; this is probably due to an increased respiratory activity in the cells grown in the presence of salt. Our results demonstrate that upon salt stress D hansenii responds to oxidative stress via the transcriptional activation of specific genes such as DhARO4., (Copyright © 2011 John Wiley & Sons, Ltd.)

The highly halotolerant yeast Debaryomyces hansenii when grown in the presence of 2M NaCl, increased the expression of ARO4 which is involved in the biosynthesis of aromatic amino acids. The function of the isolated gene was verified by complementation of a Saccharomyces cerevisiae null mutant, aro4Delta, restoring the specific activity of the enzyme (a 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase) to wild-type levels. DhARO4 transcript expression under high salinity was stimulated at the beginning of the exponential growth phase. As the DhARO4 promoter region presents putative GCRE and CRE sequences, its expression was evaluated under conditions of NaCl stress and amino acid starvation, showing similar expression levels for either condition. The combined effect of both stressors resulted in a further increase in transcript levels over the singly added stressors, indicating independent stimulatory events. Our results support the hypothesis that high salinity and amino acid availability are physiologically interconnected., (Copyright (c) 2006 John Wiley & Sons, Ltd.)

We have analyzed electrophoretic profiles of polypeptides extracted from various cell compartments of the yeast Debaryomyces hansenii, cultured under high osmolarity and under control conditions. We tested the effect of high concentrations of solutes with an osmotic component (sorbitol), and with osmotic and ionic components combined (NaCl or KCl). Densitometric analyses of the extracted polypeptides indicated that the stressing solutes had a differential effect on the relative concentration of total proteins as well as in proteins extracted from three subcellular compartments. Sorbitol caused a significant decrease in the concentration of various polypeptides associated with the mitochondria and the cytoplasm. By contrast, sodium ions elicited marked increases in concentration in four cytoplasmic polypeptides. KCl did not have a major effect in any of the subcellular compartments. Polypeptides were grouped as having a general osmotic response, or as having a response apparently modulated by the particular ionic environment of the growth medium. In all treatments, the number of polypeptides with an increase in their relative concentration was roughly similar to the number of polypeptides with a decrease in concentration, both relative to controls. Our results agree with previous observations on the complexity of the osmoregulatory response involving proteins whose concentration depends on the solute causing the stress. The results also indicate that subcellular compartments respond differently to stressors.