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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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- Baum, Matthew L, et al.
(författare)
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CSMD1 regulates brain complement activity and circuit development
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Ingår i: Brain, Behavior, and Immunity. - 1090-2139.
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Tidskriftsartikel (refereegranskat)abstract
- Complement proteins facilitate synaptic elimination during neurodevelopmental pruning, but neural complement regulation is not well understood. CUB and Sushi Multiple Domains 1 (CSMD1) can regulate complement activity in vitro, is expressed in the brain, and is associated with increased schizophrenia risk. Beyond this, little is known about CSMD1 including whether it regulates complement activity in the brain or otherwise plays a role in neurodevelopment. We used biochemical, immunohistochemical, and proteomic techniques to examine the regional, cellular, and subcellular distribution as well as protein interactions of CSMD1 in the brain. To evaluate whether CSMD1 is involved in complement-mediated synapse elimination, we examined Csmd1-knockout mice and CSMD1-knockout human stem cell-derived neurons. We interrogated synapse and circuit development of the mouse visual thalamus, a process that involves complement pathway activity. We also quantified complement deposition on synapses in mouse visual thalamus and on cultured human neurons. Finally, we assessed uptake of synaptosomes by cultured microglia. We found that CSMD1 is present at synapses and interacts with complement proteins in the brain. Mice lacking Csmd1 displayed increased levels of complement component C3, an increased colocalization of C3 with presynaptic terminals, fewer retinogeniculate synapses, and aberrant segregation of eye-specific retinal inputs to the visual thalamus during the critical period of complement-dependent refinement of this circuit. Loss of CSMD1 in vivo enhanced synaptosome engulfment by microglia in vitro, and this effect was dependent on activity of the microglial complement receptor, CR3. Finally, human stem cell-derived neurons lacking CSMD1 were more vulnerable to complement deposition. These data suggest that CSMD1 can function as a regulator of complement-mediated synapse elimination in the CNS during development.
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- Lipoma, Lucrecia, et al.
(författare)
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No general support for functional diversity enhancing resilience across terrestrial plant communities
- 2024
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Ingår i: Global Ecology and Biogeography. - : John Wiley & Sons. - 1466-822X .- 1466-8238.
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Forskningsöversikt (refereegranskat)abstract
- Aim: Understanding the mechanisms promoting resilience in plant communities is crucial in times of increasing disturbance and global environmental change. Here, we present the first meta-analysis evaluating the relationship between functional diversity and resilience of plant communities. Specifically, we tested whether the resilience of plant communities is positively correlated with interspecific trait variation (following the niche complementarity hypothesis) and the dominance of acquisitive and small-size species (following the mass ratio hypothesis), and for the context-dependent effects of ecological and methodological differences across studies.Location: Global.Time Period: 2004–2021.Major Taxa Studied: Vascular plants.Methods: We compiled a dataset of 69 independent sites from 26 studies that have quantified resilience. For each site, we calculated functional diversity indices based on the floristic composition and functional traits of the plant community (obtained from the TRY database) which we correlated with resilience of biomass and floristic composition. After transforming correlation coefficients to Fisher's Z-scores, we conducted a hierarchical meta-analysis, using a multilevel random-effects model that accounted for the non-independence of multiple effect sizes and the effects of ecological and methodological moderators. Results: In general, we found no positive functional diversity–resilience relationships of grand mean effect sizes. In contrast to our expectations, we encountered a negative relationship between resilience and trait variety, especially in woody ecosystems, whereas there was a positive relationship between resilience and the dominance of acquisitive species in herbaceous ecosystems. Finally, the functional diversity–resilience relationships were strongly affected by both ecological (biome and disturbance properties) and methodological (temporal scale, study design and resilience metric) characteristics. Main Conclusions: We rejected our hypothesis of a general positive functional diversity–resilience relationship. In addition to strong context dependency, we propose that idiosyncratic effects of single resident species present in the communities before the disturbances and biological legacies could play major roles in the resilience of terrestrial plant communities.
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- Li, Shanghuo, et al.
(författare)
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The ALMA Survey of 70 μm Dark High-mass Clumps in Early Stages (ASHES). VII. Chemistry of Embedded Dense Cores
- 2022
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Ingår i: Astrophysical Journal. - : American Astronomical Society. - 1538-4357 .- 0004-637X. ; 939:2
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Tidskriftsartikel (refereegranskat)abstract
- We present a study of the chemistry toward 294 dense cores in 12 molecular clumps, using data obtained from the ALMA Survey of 70 μm dark High-mass clumps in Early Stages. We identified 97 protostellar cores and 197 prestellar core candidates, based on the detection of outflows and molecular transitions of high upper-energy levels (E u /k > 45 K). The detection rate of the N2D+ emission toward the protostellar cores is 38%, which is higher than 9% for the prestellar cores, indicating that N2D+ does not exclusively trace prestellar cores. The detection rates of the DCO+ emission are 35% for the prestellar cores and 49% for the protostellar cores, which are higher than those for N2D+, implying that DCO+ appears more frequently than N2D+ in both prestellar and protostellar cores. Both the N2D+ and DCO+ abundances appear to decrease from the prestellar to the protostellar stage. The DCN, C2D, and 13CS emission lines are rarely seen in the dense cores of early evolutionary phases. The detection rate of the H2CO emission toward dense cores is 52%, three times higher than that for CH3OH (17%). In addition, the H2CO detection rate, abundance, line intensities, and line widths increase with the core evolutionary status, suggesting that the H2CO line emission is sensitive to protostellar activity.
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