Combined effects of fasting and vinblastine treatment on serum insulin level, the size of autophagic-lysosomal compartment, protein content and lysosomal enzyme activities of liver and exocrine pancreatic cells of the mouse

Kovács, Attila; Lénárd, László; Fellinger, Elizabeth; Jakab, Andrea Emese; Orosz, Antal; Réz, Gábor; Kovács, János

doi:10.1016/0305-0491(89)90189-2

Cited by 12 publications

(10 citation statements)

References 17 publications

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“…Examples of actual morphometric measurements for the characterization of autophagic processes can be found in several articles. 21,108,111,115,116 It is appropriate to note here that a change in the volume fraction of the autophagic compartment may come from 2 sources; from the real growth of its size in a given cytoplasmic volume, or from the decrease of the cytoplasmic volume itself. To avoid this so-called "reference trap," the reference space volume can be determined by different methods.…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

“…A catalytic MTOR inhibitor that increases macroautophagic flux to a greater level than allosteric inhibitors such as rapamycin (and may be used to induce macroautophagy in cell lines that are resistant to rapamycin and its derivatives); 106,112,116,186,192,193,195,196,204,206,207,210,211,212,214,216,218 E Electron microscopy 30,34,35,61,84,117,121 Endosomal microautophagy 96,196,200 EPG 69,105,189,190,197,204,212,213,214,216,217 109,110,111,113,114,115,116,121,187,190,195,…”

Section: Wye-354mentioning

confidence: 99%

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Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Klionsky¹,

Abdelmohsen²,

Abe³

et al. 2016

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Section: Transmission Electron Microscopymentioning

confidence: 99%

Section: Wye-354mentioning

confidence: 99%

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Klionsky¹,

Abdelmohsen²,

Abe³

et al. 2016

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“…The best approach is to estimate the volume occupied by autophagic structures (as percent of cytoplasmic or cellular volume, using volumetric morphometry/stereology) in at least 20 cell profiles per sample (the number needed should be dictated by some form of power analysis, which indicates that the data are significant). 20,65,68,70,71 During quantification it is important to make sure that each imaged cell profile is captured and scored at the same magnification, and that every cell profile in the thin section has an equal probability to be included in the counting.…”

Section: Introductionmentioning

confidence: 99%

Guidelines for the use and interpretation of assays for monitoring autophagy

Klionsky¹,

Abdalla²,

Abeliovich³

et al. 2012

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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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“…17 The accumulation of autophagic vacuoles was influenced by the feeding status of the animals after vinblastine treatment, which also caused the decrease of the serum insulin levels. 16 These results point out the need for careful consideration of the possible multiple interactions of various substances and regulating factors to influence autophagy. This message is becoming more important again when in vivo studies of autophagy are gaining more ground.…”

Section: Djkmentioning

confidence: 99%

“…Morphometry also opened up the way to expand our work toward comparative studies of different tissues involving liver cells. [14][15][16][17][18][19] This was all the more important since practically no biochemical data were available in the literature for tissues other than the liver. Logically, therefore, while working with the tissue slices of the seminal vesicle, I also looked for in vitro methods to study liver cells.…”

mentioning

confidence: 99%