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

Amorim Neto, Dionisio Pedro; de Castro Fonseca, Matheus

Amorim Neto, Dionisio Pedro; de Castro Fonseca, Matheus.

Afiliação

Amorim Neto DP; Department of Food Science and Nutrition, Faculty of Food Engineering, University of Campinas, Campinas, Brazil.
de Castro Fonseca M; Division of Biology and Biological Engineering, California Institute of Technology, 1200 E. California Boulevard, Pasadena, CA, USA. Electronic address: mdecastr@caltech.edu.

STAR Protoc ; 3(3): 101486, 2022 09 16.

Article em En | MEDLINE | ID: mdl-35755129

RESUMO

Gut microbiota influences neurodevelopment, behavior and contributes to neurodegenerative disorders. One possible mechanism is the direct modulation of calcium (Ca2+) signaling and protein homeostasis in enteroendocrine cells (EECs), a component of the gut epithelium. Here, we present a protocol to isolate fractions of conditioned media (CM) from the anaerobic bacteria Akkermansia muciniphila and the utilization of this CM to monitor Ca2+ fluctuation in EECs by imaging. 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).

Assuntos

Bactérias Anaeróbias; Sinalização do Cálcio; Anaerobiose; Meios de Cultivo Condicionados/farmacologia; Células Enteroendócrinas

Palavras-chave

Cell Biology; Microbiology; Microscopy

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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bactérias Anaeróbias / Sinalização do Cálcio Idioma: En Revista: STAR Protoc Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos

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