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) : Edp Sciences S A
Résumé : Sperm characteristics of scallops have not been well described in the scientific literature. The effects of sperm release technique (thermal shock versus serotonin injection), of sperm collection technique (testis sampling versus serotonin injection), of sperm sampling location along the genital tract, of in vitro sperm maturation, and of time post activation on scallop sperm characteristics were assessed in the present work. Whatever sperm release technique used, no significant differences were observed regarding the percentage of motile spermatozoa and the velocity of the average path (YAP). Compared to testicular sperm, a higher percentage of motile spermatozoa, YAP and intracellular adenosine triphosphate (ATP) content were observed for sperm shed after serotonin injection. From the distal part of testes up to the gonopore, an increase of the percentage of motile spermatozoa and YAP was assessed, suggesting a sperm 'maturation process' along the genital ducts. A higher increase in the percentage of motile sperm was recorded during a 5 mm incubation of testicular sperm in seawater containing 2 mM serotonin and seawater containing 10 mM caffein compared to seawater (control). In addition, a higher YAP was assessed, incubating testicular sperm in caffein, compared to control or serotonin. Then, the percentage of motile spermatozoa, YAP and intracellular ATP content exhibited a progressive reduction during the 10 h swimming period. Mean values of the percentage of motile spermatozoa, YAP, sperm track linearity (LIN) and intracellular ATP content recorded at the beginning of the movement period for sperm samples collected after intragonadal serotonin injection, were 82 +/- 7%, 162 +/- 15 mu m s(-1), 0.33 +/- 0.12 and 212 +/- 133 nmol x 10(-9) spermatozoa (n = 9 males), respectively. The present study confirms the existence of a sperm "maturation process" along scallop genital ducts. In addition, the cessation of scallop sperm movement can be explained by the exhaustion of ATP content at the end of the movement phase.
Aquatic Living Resources (0990-7440) (Edp Sciences S A), 2013-07 , Vol. 26 , N. 3 , P. 215-220

Droits : EDP Sciences, IFREMER, IRD 2013
http://archimer.ifremer.fr/doc/00157/26852/24983.pdf
DOI:10.1051/alr/2013048
http://archimer.ifremer.fr/doc/00157/26852/

Éditeur(s) : 3e Rencontres Scientifiques Internationales du contrat de baie de la rade de Brest. Brest 14-16 mars 1995
Résumé : Aquaculture production of scallop has been identified by a series of technical operations during the past ten years programme under a partnership involving the fishermen organizations with regional and national grants. 3 million juveniles have been produced annually, giving a potential recapture by fishing of 100 to 120 tonnes 3 years later. Results in terms of technical and economical approach have pointed out the principal unknown factors responsable for high costs. 7he pilot programme aims to increase the production to reach a level of 5 million juveniles to support the fishery of the rade de Brest and induce new management of the resources. Two priorities were selected, control of the reproduction in the hatchery to obtain reliable larval and postlarval rearing during a larger period of the year, mainly during summer, and improvement of survival rate of the juveniles by minimizing stress when manipulating the animals. But biological and ecological mechanisms are still poorly understoodfor scallop and generally bibvalves and the last two years have shown unchanged results. The technical pathways shows interactions between the different steps of the production of juveniles where different teams operate separately. Management of the livestock has to be developed to increase the survival rate by better breeding but also to reduce the loss by adapted and coordinated practices. Recapture results indicate that fishery strategies depend also on the production of juvenile, and could orientate the management of the hatchery.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00044/15494/12860.pdf
http://archimer.ifremer.fr/doc/00044/15494/

Éditeur(s) : Elsevier Science Inc
Résumé : The sex-determining system of Crassostrea gigas is still poorly known, especially regarding the potential influence of temperature. In order to address this question, mRNA expressions of actors of the molecular cascade (Cg-DMI, Cg-SoxE, Cg-beta-catenin, Cg-Foxl2/Cg-FoxI2os) and of Oyvlg, a germ cell marker, were investigated by real-time PCR in spat grown at different temperatures (18, 22, 25 and 28 degrees C). In parallel, gonadic differentiation, gametogenesis and sex ratios were assessed by histology at each of these temperatures. Whatever the temperature, Cg-DMI, Cg-SoxE, Cg-beta-catenin and Oyvlg expressions peaked at the same developmental stage, always after Cg-Foxl2/Cg-Foxl2os (around 40-44 dpf for spat grown at 18 degrees C). Temperatures increased the kinetics of first gonadic differentiation and gametogenesis. At 25 degrees C a significant switch occurred in sex ratio towards males and in the balance of expression between male and female genes, in favor of males. A slight gametogenesis disturbance was also observed. These results strengthen the hypotheses about the sex-determining time window and molecular cascade governing the development of C gigas, with notably the involvement of Cg-Foxl2/Cg-Foxl2os in the very early steps. They also suggest an influence of temperature on the oyster's sex determination which, associated to genetic control, would induce a mixed sex determination system (GSD +TSD). (C) 2013 Elsevier Inc. All rights reserved.
Comparative Biochemistry And Physiology A-molecular & Integrative Physiology (1095-6433) (Elsevier Science Inc), 2013-05 , Vol. 165 , N. 1 , P. 61-69

Droits : 2013 Elsevier Inc. All rights reserved
http://archimer.ifremer.fr/doc/00136/24767/22874.pdf
DOI:10.1016/j.cbpa.2013.02.007
http://archimer.ifremer.fr/doc/00136/24767/