斑马鱼Danio rerio肽转运体1a,营养基因组学和营养研究中的新兴模型:分子表征,功能特性和表达分析。

Genes & Nutrition Pub Date : 2019-12-19 eCollection Date: 2019-01-01 DOI:10.1186/s12263-019-0657-3
Francesca Vacca, Amilcare Barca, Ana S Gomes, Aurora Mazzei, Barbara Piccinni, Raffaella Cinquetti, Gianmarco Del Vecchio, Alessandro Romano, Ivar Rønnestad, Elena Bossi, Tiziano Verri
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引用次数: 11

摘要

背景:肽转运蛋白1 (PepT1,别名Slc15a1)介导所有脊椎动物对膳食二肽/三肽的摄取。然而,在硬骨鱼中,由于在进化过程中发生的特定全基因组复制事件导致比高等脊椎动物(四足动物)更复杂的旁系基因库,因此可能有不止一种pept1型转运蛋白起作用。结果:在这里,我们描述了斑马鱼(Danio rerio)中一种新的二/三肽转运蛋白,即斑马鱼肽转运蛋白1a (PepT1a;也被称为溶质载体家族15成员a1, Slc15a1a),它是前面描述的斑马鱼肽转运体1b (PepT1b,别名PepT1;也称为溶质载体家族15成员1b, Slc15a1b)。此外,我们报告了斑马鱼成年组织/器官和胚胎/早期幼虫发育阶段pept1a (slc15a1a) mRNA表达水平的基本分析。通过在非洲爪鱼卵母细胞中的表达和双电极电压钳测量来评估,斑马鱼PepT1a,即PepT1b,具有电致性、Na+无关性和pH依赖性,是一个低亲和系统,gy - gln在- 60 mV时的K值为6.92 mmol/L, pH值为7.6和0.24 mmol/L,在- 120 mV时的K值为3.61 mmol/L, pH值为7.6和0.45 mmol/L。斑马鱼pept1a mRNA在成年鱼的肠道和卵巢中高度表达,但其在发育早期的表达经历了一个复杂的趋势,在受精后1和2天(dpf)检测到pept1a mRNA,可能是由于它出现在RNA母池中,在3 dpf时下降(约0.5倍),在4至7 dpf时增加1-2 dpf,在6 dpf时达到峰值(约7倍)。结论:我们发现斑马鱼pept1a型转运体在成年鱼肠中与pept1b (slc15a1b)共表达。在卵黄合胞层存在和卵黄蛋白吸收过程活跃的发育早期,也证实了它的表达。这些结果填补了斑马鱼中pept1型转运蛋白功能的缺失信息,为进一步研究PepT1a/PepT1b在斑马鱼和硬骨鱼生理中的相似/差异作用开辟了道路。
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The peptide transporter 1a of the zebrafish Danio rerio, an emerging model in nutrigenomics and nutrition research: molecular characterization, functional properties, and expression analysis.

Background: Peptide transporter 1 (PepT1, alias Slc15a1) mediates the uptake of dietary di/tripeptides in all vertebrates. However, in teleost fish, more than one PepT1-type transporter might function, due to specific whole genome duplication event(s) that occurred during their evolution leading to a more complex paralogue gene repertoire than in higher vertebrates (tetrapods).

Results: Here, we describe a novel di/tripeptide transporter in the zebrafish (Danio rerio), i.e., the zebrafish peptide transporter 1a (PepT1a; also known as Solute carrier family 15 member a1, Slc15a1a), which is a paralogue (78% similarity, 62% identity at the amino acid level) of the previously described zebrafish peptide transporter 1b (PepT1b, alias PepT1; also known as Solute carrier family 15 member 1b, Slc15a1b). Also, we report a basic analysis of the pept1a (slc15a1a) mRNA expression levels in zebrafish adult tissues/organs and embryonic/early larval developmental stages. As assessed by expression in Xenopus laevis oocytes and two-electrode voltage clamp measurements, zebrafish PepT1a, as PepT1b, is electrogenic, Na+-independent, and pH-dependent and functions as a low-affinity system, with K 0.5 values for Gly-Gln at - 60 mV of 6.92 mmol/L at pH 7.6 and 0.24 mmol/L at pH 6.5 and at - 120 mV of 3.61 mmol/L at pH 7.6 and 0.45 mmol/L at pH 6.5. Zebrafish pept1a mRNA is highly expressed in the intestine and ovary of the adult fish, while its expression in early development undergoes a complex trend over time, with pept1a mRNA being detected 1 and 2 days post-fertilization (dpf), possibly due to its occurrence in the RNA maternal pool, decreasing at 3 dpf (~ 0.5-fold) and increasing above the 1-2 dpf levels at 4 to 7 dpf, with a peak (~ 7-fold) at 6 dpf.

Conclusions: We show that the zebrafish PepT1a-type transporter is functional and co-expressed with pept1b (slc15a1b) in the adult fish intestine. Its expression is also confirmed during the early phases of development when the yolk syncytial layer is present and yolk protein resorption processes are active. While completing the missing information on PepT1-type transporters function in the zebrafish, these results open to future investigations on the similar/differential role(s) of PepT1a/PepT1b in zebrafish and teleost fish physiology.

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