Éditeur(s) : HAL CCSD Landes Bioscience
Résumé : International audience
In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
Autophagy

hal-00735751
https://hal.archives-ouvertes.fr/hal-00735751
PUBMED : 22966490

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Kimberlites of the Udachnaya-East pipe (Siberia) include a uniquely dry and serpentine-free rock type with anomalously high contents of chlorine (Cl ≤ 6.1 wt%), alkalies (Na2O + K2O ≤ 10 wt%) and sulfur (S ≤ 0.50 wt%), referred to as a “salty” kimberlite. The straightforward interpretation is that the Na-, K-, Cl- and S-rich components originate directly from a carbonate-chloride kimberlitic magma that is anhydrous and alkali-rich. However, because brines and evaporites are present on the Siberian craton, previous studies proposed that the kimberlitic magma was contaminated by the assimilation of salt-rich crustal rocks. To clarify the origin of high Cl, alkalies and S in this unusual kimberlite, here we determine its sulfur speciation and isotopic composition and compare it to that of non-salty kimberlites and kimberlitic breccia from the same pipe, as well as potential contamination sources (hydrothermal sulfides and sulfates, country-rock sediment and brine collected in the area). The average δ34S of sulfides is − 1.4 ± 2.2‰ in the salty kimberlite, 2.1 ± 2.7‰ in the non-salty kimberlites and 14.2 ± 5.8‰ in the breccia. The average δ34S of sulfates in the salty kimberlites is 11.1 ± 1.8‰ and 27.3 ± 1.6‰ in the breccia. In contrast, the δ34S of potential contaminants range from 20 to 42‰ for hydrothermal sulfides, from 16 to 34‰ for hydrothermal sulfates, 34‰ for a country-rock sediment (Chukuck suite) and the regional brine aquifer. Our isotope analyses show that (1) in the salty kimberlites, neither sulfates nor sulfides can be simply explained by brine infiltration, hydrothermal alteration or the assimilation of known salt-rich country rocks and instead, we propose that they are late magmatic phases; (2) in the non-salty kimberlite and breccia, brine infiltration lead to sulfate reduction and the formation of secondary sulfides – this explains the removal of salts, alkali-carbonates and sulfates, as well as the minor olivine serpentinization; (3) hydrothermal sulfur was added to the kimberlitic breccia, but not to the massive kimberlites. In situ measurements of sulfides confirm this scenario, clearly showing the addition of two sulfide populations in the breccia (pyrite-pyrrhotites with average δ34S of 7.9 ± 3.4‰ and chalcopyrites with average δ34S of 38.0 ± 0.4‰) whereas the salty and non-salty kimberlites preserve a unique population of djerfisherites (Cl- and K-rich sulfides) with δ34S values within the mantle range. This study provides the first direct evidence of alkaline igneous rocks in which magmatic sulfate is more abundant than sulfide. Although sulfates have been rarely reported in mantle materials, sulfate-rich melts may be more common in the mantle than previously thought and could balance the sulfur isotope budget of Earth's mantle.
ISSN: 0009-2541

hal-01571982
https://hal.archives-ouvertes.fr/hal-01571982
DOI : 10.1016/j.chemgeo.2016.10.037