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Overington, E.

Publications and source records attributed to Overington, E..

2 recordsLinked to original sources

Protease-activated receptor 1-mediated matrix metalloprotease signalling in sensory neurons

Visceral pain is a prevalent and debilitating symptom of inflammatory bowel disease (IBD). However, current pain therapies are often ineffective, raising the possibility that novel disease mediators might be contributing to pain during inflammation. Our study provides new insights into how matrix metalloproteinases (MMPs), which are elevated in IBD, stimulate sensory neurons. We demonstrate that MMP3, MMP8, and MMP9 induce intracellular Ca2+ release in dorsal root ganglion (DRG) neurons through activation of protease-activated receptor 1 (PAR1) and subsequent activation of phospholipase C (PLC). Characterisation of the neuronal populations stimulated by these MMPs suggests that a subset is likely nociceptive. In contrast, MMP2 and MMP13, although capable of cleaving PAR1 in other cell types, do not induce Ca2+ mobilisation in DRG neurons. Interestingly, pre-treatment with MMP2 or MMP13 reduces the neuronal response to MMP3 or PAR1 agonist, suggesting that MMP2 and MMP13 act on PAR1 in a manner which prevents further activation. Additionally, MMP10 induces Ca2+ mobilisation in DRG neurons but through a PAR1-independent mechanism. These findings uncover a previously unrecognised role for MMP signalling in sensory neurons, highlighting a potential mechanism by which MMPs could contribute to the pro-nociceptive environment in the inflamed bowel.

neuroscience↗

Mechanisms of Activation and Serotonin release from Human Enterochromaffin Cells

Background and AimsGastrointestinal (GI) enterochromaffin (EC) cells are specialised sensors of luminal stimuli. They secrete most of the bodys serotonin (5-HT), and are critical for modulating GI motility, secretion, and sensation, while also signalling satiety and intestinal discomfort. The aim of this study was to investigate mechanisms underlying the regulation of human EC cells, and the relative importance of direct nutrient stimulation compared with neuronal and paracrine regulation. MethodsIntestinal organoids from human duodenal biopsies were modified using CRISPR-Cas9 to specifically label EC cells with either the fluorescent protein Venus or the cAMP sensor Epac1-S. EC cells were purified by fluorescence-activated cell sorting for analysis by bulk RNA sequencing and liquid chromatography mass spectrometry peptidomics. The function of human EC cells was studied using single cell patch clamp, calcium and cAMP imaging and 5-HT ELISA assays. ResultsHuman EC cells showed expression of receptors for nutrients (including GPR142, GPBAR1, GPR119, FFAR2, OR51E1, OR51E2), gut hormones (including SSTR1,2&5, NPY1R, GIPR) and neurotransmitters (ADRA2A, ADRB1). Functional assays revealed EC responses (calcium, cAMP and/or secretion) to a range of stimuli, including bacterial metabolites, aromatic amino acids and adrenergic agonists. Electrophysiological recordings showed that isovalerate increased action potential firing. Conclusions5-HT release from EC cells controls many physiological functions and is currently being targeted to treat disorders of the gut-brain axis. Studying ECs from human organoids enables improved understanding of the molecular mechanisms underlying EC cell activation, which is fundamental for the development of new strategies to target 5-HT-related gut and metabolic disorders. SynopsisHuman duodenal organoids expressing fluorescent proteins in enterochromaffin cells were used to study mechanisms underlying serotonin secretion. Different expression of key sensory receptors was identified by transcriptomic analysis, and validated by live cell second messenger imaging and secretion assays.

cell biology↗