Gastrointestinal (GI) enterochromaffin (EC) cells are specialised sensors of luminal stimuli. They secrete most of the body's serotonin (5-HT), and are critical for modulating GI motility, secretion, Show more
Gastrointestinal (GI) enterochromaffin (EC) cells are specialised sensors of luminal stimuli. They secrete most of the body's serotonin (5-HT), and are critical for modulating GI motility, secretion, and sensation, while also signaling 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. Intestinal organoids from human duodenal biopsies were modified using CRISPR-Cas9 to specifically label EC cells with either the fluorescent protein Venus or the cyclic adenosine monophosphate (cAMP) sensor Epac1-S-H187. 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-hydroxytryptamine (5-HT) enzyme-linked immunosorbent assays (ELISAs). Human 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. 5-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. Show less
Single nucleotide polymorphism (SNP) in the A total of 229 hyperlipidaemic statin users (96.1% Malays) were recruited, and a single blood sample (3 mL) was obtained for DNA extraction. The genotypes w Show more
Single nucleotide polymorphism (SNP) in the A total of 229 hyperlipidaemic statin users (96.1% Malays) were recruited, and a single blood sample (3 mL) was obtained for DNA extraction. The genotypes were determined using PCR-RFLP method and validated by sequencing analysis. The minor allele frequency for rs708272 in all participants was 0.391, with no difference between females and males. At the baseline, the SNP was associated with different low-density lipoprotein (LDL-c) and triglyceride (TG) levels in females, but not males, when the GG and GA+AA genotypes were compared using a dominant genetic model. Regardless of the genotype, the total cholesterol and LDL-c levels decreased significantly ( To improve the management of hyperlipidaemia, future research should consider patient gender when assessing the Show less