Uptake of manganese from manganese–lysine complex in the primary rat intestinal epithelial cells

Abstract

This study was conducted to compare the differences of the uptake of Mn from Mn-lysine complex (MnLys) and MnSO and to determine the potential mechanisms of the uptake of Mn from MnLys. We first established the primary rat intestinal epithelial cell culture model and used it to determine the uptake of Mn from different Mn sources. The MnLys increased (p < 0.001) Mn uptake when compared to MnSO . The uptake of Mn decreased (p < 0.05) with added Fe concentration increasing in the medium regardless of Mn source. The MnLys decreased (p < 0.01) Mn efflux transporter ferroportin 1 (FPN1) mRNA level, but did not influence (p > 0.06) Mn influx transporter DMT1 mRNA expression when compared to MnSO . The results above indicated that the increase of Mn accumulation for MnLys at least partly was due to the decrease of Mn efflux by reduced FPN1 expression. The N-ethylmaleimide, as an l-lysine transport system y inhibitor, decreased (p < 0.001) the uptake of Mn from MnLys, but did not affect (p > 0.10) the uptake of Mn from MnSO . The cycloheximide, as an l-lysine transport system b activator, increased (p < 0.001) the uptake of Mn from MnLys, whereas also did not influence the uptake of Mn from MnSO . The MnLys increased (p < 0.01) the system y member cationic amino acid transporter (CAT) 1, and system b components rBAT and b AT mRNA expression when compared to MnSO . These results suggested that the uptake of Mn from MnLys complex might be transported by CAT1 and system b , which was different from ionized Mn uptake pathway. In conclusion, the uptake of MnLys complex not only might be absorbed as Mn , but also appeared to be transported through CAT1 and system b in the primary rat intestinal epithelial cells.

Effect of zinc sources and experimental conditions on zinc balance in growing wethers

“…Although Nockels et al (1993) observed decreases in Zn absorption during stress, no difference in absorption was observed between different sources of Zn (ZS and Zn-Met). The lower retention of Zn from ZG as compared with ZAA might be true based on the theory that trace minerals bound to amino acids are absorbed into the enterocyte via amino acids transport system vs. transporters such as ZIP4 ( Gao et al, 2014 ; Sauer et al, 2017 ; Zhang et al, 2017 ) and that Gly is less efficiently absorbed than other amino acids ( Gardner, 1976 ; Reichl, 1989 ). Further research is needed to explore differences in Zn metabolism among Zn sources as affected by factors such as stress, animal type (monogastric vs. ruminant), and ligand.…”

Uptake of Manganese from the Manganese-Lysine Complex in Primary Chicken Intestinal Epithelial Cells

“…And then chicken IECs proliferated to a monolayer of cuboidal cells at 3–4 days post-seeding. The morphological characteristics of chicken IECs were similar to the primary rat IECs but the time for forming confluence monolayer was earlier in primary chicken IECs than in primary rat IECs [33]. The presence of cytokeratin-18, an intermediate cytoskeletal filament that traverses the cell cytoplasm, is characteristic of IECs [37], and vimentin is an intermediate filament specific for intestinal fibroblast [40].…”

“…Through this method, cells were assessed for the presence of cobblestone morphology, crypts, and the degree of fibroblast contamination. Cytokeratin-18 (an intermediate filament specific for epithelial cells) and vimentin (an intermediate filament specific for mesenchymal cells such as intestinal fibroblast) were visualized by IF staining [33]. Cells were fixed with ice-cold methanol for 15 min, subsequently washed with PBST (PBS plus 0.05% Tween 20), and blocked for 45 min with 5% skim milk powder at room temperature (RT).…”

“…For time-dependent Mn uptake assay, the uptake was initiated by adding 2 mL this uptake buffer containing 0.25 mM Mn from MnSO 4 . Uptake of Mn was arrested by discarding the uptake buffer at 0, 15, 15, 30, 45, 60, 120, 180, or 240 min post-treatment [33] and washing cells three times with ice-cold uptake buffer. For dose-dependent Mn uptake assay, the uptake was initiated by adding 1 mL uptake buffer containing 0, 0.125, 0.25, 0.50, 0.75, or 1.00 mM Mn from MnSO 4 , and Mn uptake was determined during the linear uptake period (60 min post-treatment).…”

Leveraging machine learning to dissect role of combinations of amino acids in modulating the effect of zinc on mammalian cell growth

“…Many trace metals such as zinc, manganese, copper ions can be transported into the cells as free ions or transported as a complex with amino acids such as histidine, lysine and methionine through amino acid transporters. [14][15][16][17][18][19][20][21] The uptake rate via these two mechanisms are however different. 22 Amino acid concentration in the medium can thus affect the bioavailability and toxicity of trace metals.…”