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 Springer Verlag
Résumé : International audience
Transverse drying shrinkage was measured at microscopic and mesoscopic levels in poplar wood characterised by an increasing growth strain (GS), from normal to tension wood. Results show that: (a) The drying shrinkage, measured as a relative thickness decrease, was significantly higher for G-layer (GL) than the other layers (OL), GL shrinkage was not significantly correlated with GS and OL shrinkage was negatively correlated with GS. (b) In gelatinous fibre (G-fibre), lumen size increased during drying and this increase was positively related with GS, but in normal wood fibre lumen size decreased during drying. These findings suggest that GL shrank outwards, so that its shrinkage feebly affected the total cell shrinkage and the mesoscopic shrinkage was controlled by the OL shrinkage which shrank inwards. (c) Measurements done on 7×7 mm² thin sections evidenced a negative correlation between transverse shrinkage and GS, significant in T direction but weak in R direction. These observations at both levels allow to discuss the contribution of GL to the mesoscopic shrinkage of tension wood.
ISSN: 0043-7719

hal-00194930
https://hal.archives-ouvertes.fr/hal-00194930
https://hal.archives-ouvertes.fr/hal-00194930/document
https://hal.archives-ouvertes.fr/hal-00194930/file/WST_FANG2006.pdf
DOI : 10.1007/s00226-007-0148-3