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

Shiping Bai,Zuowei Su,Jianping Wang,Jie Bai,Xuemei Ding,Qiufeng Zeng,Yue Xuan,Bin Wu,Keying Zhang,Huanwei Peng

doi:10.3390/ani9080559

Abstract

Simple SummaryManganese (Mn) supplementation is especially necessary to avian species because the absorption of dietary Mn is relatively inefficient in birds. Recently, there has been increasing interest in the use of organic Mn to replace inorganic Mn as dietary Mn supplements in poultry. This study compared the uptake of Mn from Mn-lysine complex (MnLys) and MnSO4 in the primary chicken intestinal epithelial cells when the Fe, N-ethylmaleimide (a transport system y+ inhibitor), or cycloheximide (a transport system b0,+ activator) added in the culture medium. The results revealed that the uptake of Mn from the MnLys complex not only might be transported through the ionized Mn2+ pathway, but also appeared to be transported through the transport systems y+ and b0,+ in the intestine of chickens.Organic manganese (Mn) sources can replace inorganic Mn as dietary Mn supplements in poultry. To compare the uptake of Mn from the Mn-lysine complex (MnLys) and MnSO4, we first established the primary chicken intestinal epithelial cells (IECs) model and used it to determine Mn uptake. The MnLys increased the uptake of Mn compared to MnSO4. The uptake of Mn decreased in the IECs with Fe addition in the medium regardless of the Mn sources. The MnLys decreased the Mn2+ efflux transporter ferroportin 1 (FPN1) mRNA level but did not influence the Mn2+ influx transporter divalent metal transporter 1 (DMT1) mRNA expression when compared to MnSO4. 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 addition of N-ethylmaleimide, an L-lysine transport system y+ inhibitor, decreased the uptake of Mn from MnLys but did not affect the uptake of Mn from MnSO4. The cycloheximide, as an L-lysine transport system b0,+ activator, increased the uptake of Mn from MnLys, whereas they did not influence the uptake of Mn from MnSO4. The MnLys increased the system y+ members cationic amino acid transporter (CAT) 1 and CAT2, and system b0,+ components rBAT and b0,+AT mRNA expression when compared to MnSO4. These results suggested that the uptake of MnLys complex might be transported by CAT1/2 and system b0,+, which was different from the ionized Mn2+ uptake pathway. In conclusion, the uptake of Mn from MnLys complex not only might be uptake through the ionized Mn2+ pathway, but also appeared to be transported through the CAT1/2 and system b0,+ in primary chicken IECs.

Highlights

Manganese (Mn) is necessary for normal homeostatic processes controlling reproduction, the formation of connective tissue and bone, carbohydrate, lipid metabolism, and brain function [1].The utilization of Mn has been an increasing concern because of its low efficiency of absorption from foods to animals [2,3]
Using a combination of dispase and collagenase, chicken duodenal intestinal epithelial cells (IECs) were isolated from the underlying basement membrane in clusters (Figure 1A)
The primary chicken IECs grew as a monolayer from the explants and exhibited an epithelial morphology with polygonal shape and cobblestone appearance at 3 to 4 days post-seeding

Summary

Introduction

Manganese (Mn) is necessary for normal homeostatic processes controlling reproduction, the formation of connective tissue and bone, carbohydrate, lipid metabolism, and brain function [1].The utilization of Mn has been an increasing concern because of its low efficiency of absorption from foods to animals [2,3]. Manganese (Mn) is necessary for normal homeostatic processes controlling reproduction, the formation of connective tissue and bone, carbohydrate, lipid metabolism, and brain function [1]. Several organic Mn forms, including metal amino acid complexes, have been developed as dietary Mn supplements in poultry. Mn-amino acid complex was more bioavailable than inorganic Mn sulfate in ligated chicken duodenal loop [4,5] and Caco-2 cell models [6]. Several studies reported no differences in the Mn bioavailability between organic and inorganic Mn sources [7,8]. The differences in the ligands of organic Mn forms and the evaluation criterion might partly explain the discrepancies in relative bioavailability estimates among the research reports. It is necessary to compare the uptake of Mn from manganese-lysine complex (MnLys) or Mn sulfate (MnSO4 )

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