Gastric Serotonin Biosynthesis and Its Functional Role in L-Arginine-Induced Gastric Proton Secretion

Among mammals, serotonin is predominantly found in the gastrointestinal tract, where it has been shown to participate in pathway-regulating satiation. For the stomach, vascular serotonin release induced by gastric distension is thought to chiefly contribute to satiation after food intake. However, little information is available on the capability of gastric cells to synthesize, release and respond to serotonin by functional changes of mechanisms regulating gastric acid secretion. We investigated whether human gastric cells are capable of serotonin synthesis and release. First, HGT-1 cells, derived from a human adenocarcinoma of the stomach, and human stomach specimens were immunostained positive for serotonin. In HGT-1 cells, incubation with the tryptophan hydroxylase inhibitor p-chlorophenylalanine reduced the mean serotonin-induced fluorescence signal intensity by 27%. Serotonin release of 147 ± 18%, compared to control HGT-1 cells (set to 100%) was demonstrated after treatment with 30 mM of the satiating amino acid L-Arg. Granisetron, a 5-HT3 receptor antagonist, reduced this L-Arg-induced serotonin release, as well as L-Arg-induced proton secretion. Similarly to the in vitro experiment, human antrum samples released serotonin upon incubation with 10 mM L-Arg. Overall, our data suggest that human parietal cells in culture, as well as from the gastric antrum, synthesize serotonin and release it after treatment with L-Arg via an HTR3-related mechanism. Moreover, we suggest not only gastric distension but also gastric acid secretion to result in peripheral serotonin release.

Genotype difference in the physiological characteristics of phosphorus acquisition by wheat seedlings in alkaline soils

Phosphorus (P) in soils occurs predominately as insoluble inorganic P and organic P. However, key factors controlling P acquisition by wheat (Triticum aestivum L.) seedlings are unclear. In this study, the difference in the physiological characteristics of P acquisition in alkaline soils was investigated in wheat seedlings of two cultivars Aikang 58 and Zhoumai 22. The results indicated that the shoot P concentration of Aikang 58 was significantly higher than that of Zhoumai 22 when supplied with 0 and 70 kg/ha of pure P under field conditions. When cultured in sterile nutrition solutions with equimolar amounts of P corresponding to KH₂PO₄, Ca₃(PO₄)₂, and Ca(H₂PO₄)₂ for 6 days, the P concentration in the shoots and roots of the seedlings of Aikang 58 was significantly higher than that of Zhoumai 22. However, the P concentration of seedlings of Aikang 58 did not exhibit a significant difference than that of Zhoumai 22 when cultured in phytic acid solution. Further studies suggested that the proton secretion rate was higher, and the root phosphatase activity was significantly lower in Aikang 58 compared with those in Zhoumai 22. After 48 h of successive P starvation, the inorganic phosphate (P_i) uptake rate of Aikang 58 was significantly higher compared with that of Zhoumai 22. However, no significant differences existed in the root morphology between the two cultivars. Hence, the higher P acquisition in the wheat seedlings of Aikang 58 was attributed to a higher rate of proton secretion and a stronger capacity for P_i uptake.

Circulating aldosterone induces the apical accumulation of the proton pumping V-ATPase and increases proton secretion in clear cells in the caput epididymis

Clear cells express the vacuolar proton-pumping H+-ATPase (V-ATPase) and acidify the lumen of the epididymis, a process that is essential for male fertility. The renin-angiotensin-aldosterone system (RAAS) regulates fluid and electrolyte balance in the epididymis, and a previous study showed binding of aldosterone exclusively to epididymal clear cells (Hinton BT, Keefer DA. Steroid Biochem 23: 231–233, 1985). We examined here the role of aldosterone in the regulation of V-ATPase in the epididymis. RT-PCR showed expression of the mineralocorticoid receptor [MR; nuclear receptor subfamily 3, group C member 2 (NR3C2)] and 11-β-dehydrogenase isozyme 2 (HSD11β2) mRNAs specifically in clear cells, isolated by fluorescence-activated cell sorting from B1-enhanced green fluorescent protein (EGFP) mice. Tail vein injection of adult rats with aldosterone, 1,2-dioctanoyl-sn-glycerol (DOG), or 8-(4-chlorophenylthio)-cAMP (cpt-cAMP) induced V-ATPase apical membrane accumulation and extension of V-ATPase-labeled microvilli in clear cells in the caput epididymis but not in the cauda. V-ATPase activity was measured in EGFP-expressing clear cells using the intracellular pH (pHi)-sensing dye seminaphthorhodafluor-5F-5-(and 6)-carboxylic acid, acetoxymethyl ester acetate (SNARF-5F). Aldosterone induced a rapid increase in the rate of Na+- and bicarbonate-independent pHi recovery following an NH4Cl-induced acid load in clear cells isolated from the caput but not the cauda. This effect was abolished by concanamycin A, spironolactone, and chelerythrine but not myristoylated-protein kinase inhibitor (mPKI) or mifepristone. Thus aldosterone increases V-ATPase-dependent proton secretion in clear cells in the caput epididymis via MR/NR3C2 and PKC activation. This study, therefore, identifies aldosterone as an active member of the RAAS for the regulation of luminal acidification in the proximal epididymis.