Akkermansia muciniphila induces mitochondrial calcium overload and α -synuclein aggregation in an enteroendocrine cell line

Gut microenvironmental changes as a potential trigger in Parkinson’s disease through the gut–brain axis

“…In transgenic mouse models of PD, the gut microbiota promotes pathological alpha-synuclein accumulation, neuroinflammation, and dopaminergic neuronal degeneration, along with aggravation of motor and non-motor symptoms, which were alleviated in germ-free conditions [ 113 ]. In addition, recent research has shown that Akkermansia induces intestinal alpha-synuclein aggregation as a result of mitochondrial calcium overload in intestinal enteroendocrine cells, which were hypothesized to be an initiating site for alpha-synucleinopathy in response to environmental factors because they are connected to enteric nerves and found to have adjacent accumulation of alpha-synuclein proteins [ 118 , 119 ].…”

Protocol for rapid obtention and fractionation of anaerobic bacterial conditioned media to study calcium signaling in enteroendocrine cells

“…Therefore, understanding how the secreted bacterial elements can interfere with protein homeostasis in EECs is a new path of investigation to understand the contribution of these cells to PD pathology. This protocol describes the detailed obtention of the conditioned-medium (CM) from the anaerobic microorganism A. muciniphila which is known to modulate intracellular calcium (Ca 2+ ) signaling in EECs and induce αSyn pathology in the same cell model ( Amorim Neto et al., 2022 ). In addition, we explain here how this CM can be used to monitor Ca 2+ signaling in EECs under confocal live imaging.…”

“…This protocol was written with a focus on Akkermansia muciniphila CM. Originally the Escherichia coli CM was also fractionated and also concentrated into >3 kDa and <3 kDa fractions and used to stimulate STC-1 cells in different experimental approaches ( Amorim Neto et al., 2022 ). However, NMR analysis of the concentrated fractions was done only for BHI CM from Akkermansia muciniphila .…”

“…This protocol can be adapted and applied to various bacterial cultures and cell types. For complete details on the use and execution of this protocol, please refer to Amorim Neto et al. (2022) .…”

“…For complete details on the use and execution of this protocol, please refer to Amorim Neto et al. (2022) .…”