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Plasticity in exocytosis revealed through the effects of repetitive stimuli affect the content of nanometer vesicles and the fraction of transmitter released.

ABSTRACT: Electrochemical techniques with disk and nano-tip electrodes, together with calcium imaging, were used to examine the effect of short-interval repetitive stimuli on both exocytosis and vesicular content in a model cell line. We show that the number of events decreases markedly with repeated stimuli suggesting a depletion of exocytosis machinery. However, repetitive stimuli induce a more stable fusion pore, leading to an increased amount of neurotransmitter release. In contrast, the total neurotransmitter content inside the vesicles decreases after repetitive stimuli, resulting in a higher average release fraction from each event. We suggest a possible mechanism regarding a link between activity-induced plasticity and fraction of release.

SUBMITTER: Gu C

PROVIDER: S-EPMC6815131 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Plasticity in exocytosis revealed through the effects of repetitive stimuli affect the content of nanometer vesicles and the fraction of transmitter released.

Gu Chaoyi C Larsson Anna A Ewing Andrew G AG

Proceedings of the National Academy of Sciences of the United States of America 20190930 43

Electrochemical techniques with disk and nano-tip electrodes, together with calcium imaging, were used to examine the effect of short-interval repetitive stimuli on both exocytosis and vesicular content in a model cell line. We show that the number of events decreases markedly with repeated stimuli suggesting a depletion of exocytosis machinery. However, repetitive stimuli induce a more stable fusion pore, leading to an increased amount of neurotransmitter release. In contrast, the total neurotr ...[more]

PMID: 31570594

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Project description:The roles of bacterial extracellular vesicles (EVs) in cell-to-cell signaling are progressively being unraveled. These membranous spheres released by many living cells carry various macromolecules, some of which influence host-pathogen interactions. Bacterial EVs contain RNA, which may serve in communicating with their infected hosts. Staphylococcus aureus, an opportunistic human and animal pathogen, produces EVs whose RNA content is still poorly characterized. Here, we investigated in depth the RNA content of S. aureus EVs. A high-throughput RNA sequencing approach identified RNAs in EVs produced by the clinical S. aureus strain HG003 under different environmental conditions: early- and late-stationary growth phases, and presence or absence of a sublethal vancomycin concentration. On average, sequences corresponding to 78.0% of the annotated transcripts in HG003 genome were identified in HG003 EVs. However, only ~5% of them were highly covered by reads (≥90% coverage) indicating that a large fraction of EV RNAs, notably mRNAs and sRNAs, were fragmented in EVs. According to growth conditions, from 86 to 273 highly covered RNAs were identified into the EVs. They corresponded to 286 unique RNAs, including 220 mRNAs. They coded for numerous virulence-associated factors (hld encoded by the multifunctional sRNA RNAIII, agrBCD, psmβ1, sbi, spa, and isaB), ribosomal proteins, transcriptional regulators, and metabolic enzymes. Twenty-eight sRNAs were also detected, including bona fide RsaC. The presence of 22 RNAs within HG003 EVs was confirmed by reverse transcription quantitative PCR (RT-qPCR) experiments. Several of these 286 RNAs were shown to belong to the same transcriptional units in S. aureus. Both nature and abundance of the EV RNAs were dramatically affected depending on the growth phase and the presence of vancomycin, whereas much less variations were found in the pool of cellular RNAs of the parent cells. Moreover, the RNA abundance pattern differed between EVs and EV-producing cells according to the growth conditions. Altogether, our findings show that the environment shapes the RNA cargo of the S. aureus EVs. Although the composition of EVs is impacted by the physiological state of the producing cells, our findings suggest a selective packaging of RNAs into EVs, as proposed for EV protein cargo. Our study shedds light to the possible roles of potentially functional RNAs in S. aureus EVs, notably in host-pathogen interactions.

Dataset Information

Plasticity in exocytosis revealed through the effects of repetitive stimuli affect the content of nanometer vesicles and the fraction of transmitter released.

Publications

Plasticity in exocytosis revealed through the effects of repetitive stimuli affect the content of nanometer vesicles and the fraction of transmitter released.

Similar Datasets

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