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 IOP Publishing
Résumé : The Bureau International des Poids et Mesures (BIPM), Sevres, France, hosted the 7th International Comparison of Absolute Gravimeters (ICAG) and the associated Relative Gravity Campaign (RGC) from August to September 2005.;ICAG 2005 was prepared and performed as a metrological pilot study, which aimed:;(1) To determine the gravity comparison reference values;;(2) To determine the offsets of the absolute gravimeters; and;(3) As a pilot study to accumulate experience for the CIPM Key Comparisons.;This document presents a complete and extensive review of the technical protocol and data processing procedures. The 1st ICAG-RGC comparison was held at the BIPM in 1980-1981 and since then meetings have been organized every 4 years.;In this paper, we present an overview of how the meeting was organized, the conditions of BIPM gravimetric sites, technical specifications, data processing strategy and an analysis of the final results. This 7th ICAG final report supersedes all previously published reports.;Readings were obtained from participating instruments, 19 absolute gravimeters and 15 relative gravimeters. Precise levelling measurements were carried out and all measurements were performed on the BIPM micro-gravity network which was specifically designed for the comparison.
ISSN: 0026-1394

hal-00644529
https://hal.archives-ouvertes.fr/hal-00644529
DOI : 10.1088/0026-1394/48/5/003