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- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- 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.
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- Fukutani, A, et al.
(författare)
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Evidence for Muscle Cell-Based Mechanisms of Enhanced Performance in Stretch-Shortening Cycle in Skeletal Muscle
- 2021
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Ingår i: Frontiers in physiology. - : Frontiers Media SA. - 1664-042X. ; 11, s. 609553-
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Tidskriftsartikel (refereegranskat)abstract
- Force attained during concentric contraction (active shortening) is transiently enhanced following eccentric contraction (active stretch) in skeletal muscle. This phenomenon is called stretch-shortening cycle (SSC) effect. Since many human movements contain combinations of eccentric and concentric contractions, a better understanding of the mechanisms underlying the SSC effect would be useful for improving physical performance, optimizing human movement efficiency, and providing an understanding of fundamental mechanism of muscle force control. Currently, the most common mechanisms proposed for the SSC effect are (i) stretch-reflex activation and (ii) storage of energy in tendons. However, abundant SSC effects have been observed in single fiber preparations where stretch-reflex activation is eliminated and storage of energy in tendons is minimal at best. Therefore, it seems prudent to hypothesize that factor(s) other than stretch-reflex activation and energy storage in tendons contribute to the SSC effect. In this brief review, we focus on possible candidate mechanisms for the SSC effect, that is, pre-activation, cross-bridge kinetics, and residual force enhancement (RFE) obtained in experimental preparations that exclude/control the influence of stretch-reflex activation and energy storage in tendons. Recent evidence supports the contribution of these factors to the mechanism of SSCs, and suggests that the extent of their contribution varies depending on the contractile conditions. Evidence for and against alternative mechanisms are introduced and discussed, and unresolved problems are mentioned for inspiring future studies in this field of research.
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