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母猪从妊娠后期到泌乳高峰期乳腺中氨基酸转运体的调控。

Regulation of amino acid transporters in the mammary gland from late pregnancy to peak lactation in the sow.

作者信息

Chen Fang, Zhang Shihai, Deng Zixiao, Zhou Qiqi, Cheng Lin, Kim Sung Woo, Chen Jun, Guan Wutai

机构信息

1Department of Animal Science, South China Agricultural University, Guangzhou, 510642 China.

National Engineering Research Center for Breeding Swine Industry, Guangzhou, 510642 China.

出版信息

J Anim Sci Biotechnol. 2018 Apr 8;9:35. doi: 10.1186/s40104-018-0250-4. eCollection 2018.

Abstract

BACKGROUND

Milk protein is crucial for milk quality in sows and health of newborn piglets. Plasma amino acids (AA) in sows are important precursors for milk protein synthesis in the mammary gland. In order to study the regulation of AA transported in sow mammary glands and possible underlying mechanisms, we measured the expression of genes coding for milk proteins, AA transporter expressions, and plasma AA concentrations in sows at three different physiological stages (D-17, D1 and D17 of lactation), and then further investigated the regulation of AA transport across the cell membrane by adaptive mechanisms using pig mammary epithelial cells (PMEC) as an in vitro model. PMEC were cultured in DMEM:F12 with 4 amino acid concentrations (0 × AA complex, 1 × AA complex, 5 × AA complex, and 25 × AA complex). Classes of AA complexes evaluated in this study included neutral AAs (-Ala + -Ser + -Cys), acidic AAs (-Asp, -Glu) and neutral + basic AAs (-Ala + -Ser + -Cys + -Lys).

RESULTS

Our results indicated that mRNA expression of genes coding for milk protein (αs1-casein, αs2-casein, β-casein and κ-casein) increased significantly with the advance of physiological stage ( < 0.05), and plasma concentrations of most AAs including threonine, serine, glutamate, alanine, valine, cysteine, methionine, isoleucine and tyrosine were greater at D1 of lactation compared with D-17 and D17 of lactation ( < 0.05). Additionally, protein and gene expressions of AA transporters including excitatory AA transporter 3 (EAAT3), alanine/serine/cysteine/threonine transporter (ASCT1) and sodium-coupled neutral AA transporter 1 (SNAT2) were greater in lactating sow mammary glands compared with sow mammary glands in late pregnancy ( < 0.05). The mRNA expressions of , increased significantly in the cell mediums supplemented with 5 × and 25 × of AA complexes compared with those cells cultured in DMEM/F12 cell medium ( < 0.05). The mRNA expressions of , and also increased in EBSS cell medium compared to DMEM/F12. However, only mRNA expression of decreased when AA complex was added into EBSS ( < 0.05).

CONCLUSION

AA transportation was positively regulated in sow mammary glands with the advance of physiological stage from late pregnancy to peak of lactation and AA transporters in PMECs were adaptively regulated by changed AA concentrations.

摘要

背景

乳蛋白对母猪的乳汁质量和新生仔猪的健康至关重要。母猪血浆中的氨基酸(AA)是乳腺中乳蛋白合成的重要前体。为了研究母猪乳腺中AA转运的调节及其潜在机制,我们测定了处于三个不同生理阶段(泌乳期的D-17、D1和D17)的母猪中乳蛋白编码基因的表达、AA转运体表达以及血浆AA浓度,然后以猪乳腺上皮细胞(PMEC)作为体外模型,通过适应性机制进一步研究AA跨细胞膜转运的调节。将PMEC培养于含有4种氨基酸浓度(0×AA复合物、1×AA复合物、5×AA复合物和25×AA复合物)的DMEM:F12培养基中。本研究评估的AA复合物类别包括中性AA(-丙氨酸 + -丝氨酸 + -半胱氨酸)、酸性AA(-天冬氨酸、-谷氨酸)和中性 + 碱性AA(-丙氨酸 + -丝氨酸 + -半胱氨酸 + -赖氨酸)。

结果

我们的结果表明,乳蛋白编码基因(αs1-酪蛋白、αs2-酪蛋白、β-酪蛋白和κ-酪蛋白)的mRNA表达随生理阶段的推进而显著增加(<0.05),与泌乳期的D-17和D17相比,泌乳期D1时包括苏氨酸、丝氨酸、谷氨酸、丙氨酸、缬氨酸、半胱氨酸、蛋氨酸、异亮氨酸和酪氨酸在内的大多数AA的血浆浓度更高(<0.05)。此外,与妊娠后期的母猪乳腺相比,泌乳期母猪乳腺中AA转运体的蛋白质和基因表达,包括兴奋性AA转运体3(EAAT3)、丙氨酸/丝氨酸/半胱氨酸/苏氨酸转运体(ASCT1)和钠偶联中性AA转运体1(SNAT2)更高(<0.05)。与培养于DMEM/F12细胞培养基中的细胞相比,在补充有5×和25×AA复合物的细胞培养基中,[具体基因名称1]、[具体基因名称2]和[具体基因名称3]的mRNA表达显著增加(<0.05)。与DMEM/F12相比,在EBSS细胞培养基中,[具体基因名称1]、[具体基因名称2]和[具体基因名称3]的mRNA表达也增加。然而,当向EBSS中添加AA复合物时,只有[具体基因名称4]的mRNA表达降低(<0.05)。

结论

随着生理阶段从妊娠后期到泌乳高峰期的推进,母猪乳腺中AA转运受到正向调节,并且PMEC中的AA转运体通过改变的AA浓度受到适应性调节。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffd1/5891945/3a06e8eacbfa/40104_2018_250_Fig1_HTML.jpg

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