Effects of eccentrically and concentrically biased training on mouse muscle phenotype.

INTRODUCTION: The molecular adaptations specifically induced by different muscle contraction types have only been partially elucidated. We previously demonstrated that eccentric contractions in human quadriceps elicited proteome modifications that suggest a muscle fiber typology adaptation. We address this question in a more systematic way by examining here the effects of different running modes on the mouse muscle proteome and the muscle fiber typology. METHODS: Male adult mice (C57BL6) were randomly divided into downhill running (DHR) (quadricipital eccentrically biased contractions), uphill running (UHR) (quadricipital concentrically biased contractions), and untrained control (CONT) groups. Running groups performed five training sessions on an inclined treadmill for 75 to 135 min · d(-1), and the quadriceps muscles were dissected 96 h after the last session. Muscle protein extracts of DHR and UHR groups (n = 4/group) were subjected to a two-dimensional difference in gel electrophoresis (2D-DIGE) analysis coupled with mass spectrometry. The assessment of fiber type, size, and number was performed on the rectus femoris of the three groups (n = 6/group) using myosin heavy chain immunohistochemistry. RESULTS: In the proteomic analysis, eight spots identified as the fast myosin heavy chain isoforms exhibited a lower abundance in DHR compared with UHR (P < 0.05, t-test). In contrast, adenosine triphosphate (ATP) synthase subunit α and tubulin ß were more expressed in DHR (P < 0.05). A significant higher proportion of Type I and IIa fibers was found for DHR compared with UHR or CONT groups (P < 0.05, one-way ANOVA). CONCLUSIONS: Our data suggest that the eccentrically biased contractions in mice induced specific adaptations in protein expression and muscle fiber composition, which may reflect a more oxidative muscle phenotype. The differences in stress placed on the muscle between both trainings may be responsible for some unique adaptations resulting from the eccentrically biased training.

Secretion and maturation of conotoxins in the venom ducts of Conus textile.

The 700 or more species of cone snail attack prey by employing complex venom which can vary considerably both within species and from one species to another. Cone snail venom is remarkable for the high proportion of conotoxins with varied post-translational modifications (PTMs) and for the production of more diverse toxin scaffolds than any other known venomous animal. The venom gland, which is several times longer than its shell, is also unique in being tubular. These unusual characteristics both raise questions, and provide the opportunity for research, concerning the secretion and maturation of conotoxins along the venom duct, a process which is currently not fully understood. This research uses the two mass spectrometric techniques of isotope Coded Affinity Tagging (ICAT) and label-free quantification to study each of five portions of the venom duct of Conus textile snails from New Caledonia. Fifteen conotoxins, several with different post-translational modifications (PTMs) were identified and quantified. One hundred and forty three non-identified conotoxins were also quantified. Distinctive patterns emerged, with the largest group of conotoxins increasing, then peaking in the central-proximal part, before decreasing; whilst the second largest group peaked in the distal region, generally displaying nothing in the first parts. Conotoxins from different superfamilies were commonly found to have similar distributions. A new conotoxin, PCCSKLHDNSCCGL*, was sequenced. A comparison is made with other studies to see how the process varies in cone snails from different regions.

Identification and quantification of concentration-dependent biomarkers in MCF-7/BOS cells exposed to 17ß-estradiol by 2-D DIGE and label-free proteomics.

This paper reports the identification of biomarkers resulting from the exposure of MCF-7/BOS cells to 17ß-estradiol (E(2)). The biomarkers were identified using 2 independent and complementary techniques, 2-D DIGE/MALDI-TOF peptide mass fingerprint, and 2-D UPLC-ESI MS/MS. They were identified from the cytosolic fractions of cells treated for 24h with mitogenic concentrations of 1, 30 and 500 pM of 17ß-estradiol. Five biomarkers were up-regulated proteins, namely HSP 74, EF2, FKBP4, EF1 and GDIB and one was a down-regulated protein, namely K2C8. Three of these proteins, EF2, FKBP4 and K2C8 are implicated in a network centered on the estrogen receptors ESR1 and ESR2 as well as on AKT1. After the discovery phase, three biomarkers were selected to test the presence of estrogens using selected reaction monitoring (SRM). They were monitored using SRM after incubation of MCF-7/BOS in the presence of E(2) for confirmation or selected xenoestrogens. Daidzein, coumestrol and enterolactone induced an up-regulation of EF2 and FKPB4 proteins, while tamoxifen and resveratrol induced a down-regulation. The exposure of all phytoestrogens induced the down-regulation of K2C8. These markers form a preliminary molecular signature that can be used when testing the estrogenic activity of xenobiotics, either pure or in mixtures.