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WeightNameValue
1000 Titel
  • Determination of the autophagic flux in murine and human peripheral blood mononuclear cells
1000 Autor/in
  1. Walter, Sophia |
  2. Jung, Tobias |
  3. Herpich, Catrin |
  4. Norman, Kristina |
  5. Pivovarova-Ramich, Olga |
  6. Ott, Christiane |
1000 Erscheinungsjahr 2023
1000 LeibnizOpen
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2023-03-13
1000 Erschienen in
1000 Quellenangabe
  • 11:1122998
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2023
1000 Lizenz
1000 Verlagsversion
  • https://doi.org/10.3389/fcell.2023.1122998 |
  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10040559/ |
1000 Ergänzendes Material
  • https://www.frontiersin.org/articles/10.3389/fcell.2023.1122998/full#h13 |
1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • The autophagy lysosomal system (ALS) is crucial for cellular homeostasis, contributing to maintain whole body health and alterations are associated with diseases like cancer or cardiovascular diseases. For determining the autophagic flux, inhibition of lysosomal degradation is mandatory, highly complicating autophagy measurement in vivo. To overcome this, herein blood cells were used as they are easy and routinely to isolate. Within this study we provide detailed protocols for determination of the autophagic flux in peripheral blood mononuclear cells (PBMCs) isolated from human and, to our knowledge the first time, also from murine whole blood, extensively discussing advantages and disadvantages of both methods. Isolation of PBMCs was performed using density gradient centrifugation. To minimize changes on the autophagic flux through experimental conditions, cells were directly treated with concanamycin A (ConA) for 2 h at 37°C in their serum or for murine cells in serum filled up with NaCl. ConA treatment decreased lysosomal cathepsins activity and increased Sequestosome 1 (SQSTM1) protein and LC3A/B-II:LC3A/B-I ratio in murine PBMCs, while transcription factor EB was not altered yet. Aging further enhanced ConA-associated increase in SQSTM1 protein in murine PBMCs but not in cardiomyocytes, indicating tissue-specific differences in autophagic flux. In human PBMCs, ConA treatment also decreased lysosomal activity and increased LC3A/B-II protein levels, demonstrating successful autophagic flux detection in human subjects. In summary, both protocols are suitable to determine the autophagic flux in murine and human samples and may facilitate a better mechanistic understanding of altered autophagy in aging and disease models and to further develop novel treatment strategies.
1000 Sacherschließung
lokal TFEB
lokal cardiomyocytes
lokal lysosomes
lokal NZO
lokal sequestome-1
lokal concanamycin A
lokal LC3
lokal aging
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
  1. https://frl.publisso.de/adhoc/uri/V2FsdGVyLCBTb3BoaWE=|https://frl.publisso.de/adhoc/uri/SnVuZywgVG9iaWFz|https://frl.publisso.de/adhoc/uri/SGVycGljaCwgQ2F0cmlu|https://frl.publisso.de/adhoc/uri/Tm9ybWFuLCBLcmlzdGluYQ==|https://frl.publisso.de/adhoc/uri/UGl2b3Zhcm92YS1SYW1pY2gsIE9sZ2E=|https://frl.publisso.de/adhoc/uri/T3R0LCBDaHJpc3RpYW5l
1000 Label
1000 Förderer
  1. German Center for Cardiovascular Research |
  2. Deutsche Forschungsgemeinschaft |
1000 Fördernummer
  1. 81Z2100502
  2. FOR 2558; RA 3340/3-1, OP-R; 491394008
1000 Förderprogramm
  1. -
1000 Dateien
1000 Objektart article
1000 Beschrieben durch
1000 @id frl:6452646.rdf
1000 Erstellt am 2023-05-25T08:33:21.401+0200
1000 Erstellt von 317
1000 beschreibt frl:6452646
1000 Bearbeitet von 317
1000 Zuletzt bearbeitet Thu May 25 08:34:58 CEST 2023
1000 Objekt bearb. Thu May 25 08:34:22 CEST 2023
1000 Vgl. frl:6452646
1000 Oai Id
  1. oai:frl.publisso.de:frl:6452646 |
1000 Sichtbarkeit Metadaten public
1000 Sichtbarkeit Daten public
1000 Gegenstand von

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