TY  - JOUR
AU  - Groh, Nicole
AU  - Gallotta, Ivan
AU  - Lechler, Marie C
AU  - Huang, Chaolie
AU  - Jung, Raimund
AU  - David, Della C
TI  - Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans.
JO  - JoVE journal
VL  - Biology
IS  - 129
SN  - 1940-087X
CY  - New Delhi
PB  - JoVE124831
M1  - DZNE-2020-05990
SP  - 56464
PY  - 2017
AB  - In the last decades, the prevalence of neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), has grown. These age-associated disorders are characterized by the appearance of protein aggregates with fibrillary structure in the brains of these patients. Exactly why normally soluble proteins undergo an aggregation process remains poorly understood. The discovery that protein aggregation is not limited to disease processes and instead part of the normal aging process enables the study of the molecular and cellular mechanisms that regulate protein aggregation, without using ectopically expressed human disease-associated proteins. Here we describe methodologies to examine inherent protein aggregation in Caenorhabditis elegans through complementary approaches. First, we examine how to grow large numbers of age-synchronized C. elegans to obtain aged animals and we present the biochemical procedures to isolate highly-insoluble-large aggregates. In combination with a targeted genetic knockdown, it is possible to dissect the role of a gene of interest in promoting or preventing age-dependent protein aggregation by using either a comprehensive analysis with quantitative mass spectrometry or a candidate-based analysis with antibodies. These findings are then confirmed by in vivo analysis with transgenic animals expressing fluorescent-tagged aggregation-prone proteins. These methods should help clarify why certain proteins are prone to aggregate with age and ultimately how to keep these proteins fully functional.
KW  - Age Factors
KW  - Animals
KW  - Animals, Genetically Modified
KW  - Caenorhabditis elegans: chemistry
KW  - Caenorhabditis elegans: genetics
KW  - Caenorhabditis elegans: metabolism
KW  - Caenorhabditis elegans Proteins: chemistry
KW  - Caenorhabditis elegans Proteins: genetics
KW  - Caenorhabditis elegans Proteins: metabolism
KW  - Models, Animal
KW  - Protein Aggregates
KW  - Recombinant Proteins: chemistry
KW  - Recombinant Proteins: genetics
KW  - Recombinant Proteins: metabolism
KW  - Caenorhabditis elegans Proteins (NLM Chemicals)
KW  - Protein Aggregates (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:29286457
C2  - pmc:PMC5755480
DO  - DOI:10.3791/56464
UR  - https://pub.dzne.de/record/139668
ER  -