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Regulation of leucine catabolism by metabolic fuels in mammary epithelial cells

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Abstract

Lactation is associated with elevated catabolism of branched-chain amino acids (BCAA) in mammary glands to produce glutamate, glutamine, alanine, aspartate, and asparagine. This study determined effects of metabolic fuels on the catabolism of leucine (a representative BCAA) in bovine mammary epithelial cells. Cells were incubated at 37 °C for 2 h in Krebs buffer containing 0.5 mM l-leucine and either l-[1-14C]leucine or l-[U-14C]leucine. The medium also contained 0–5 mM d-glucose, 0–2 mM l-glutamine, 0–4 mM dl-β-hydroxybutyrate, or 0–2 mM oleic acid. Rates of leucine decarboxylation were 60 % lower, but rates of α-ketoisocaproate production were 34 % higher, in the presence of 2 mM glucose than in its absence. All variables of leucine catabolism did not differ between 2 and 5 mM glucose or between 0 and 4 mM dl-β-hydroxybutyrate. Compared with 0–0.25 mM glutamine, 0.5 and 2 mM l-glutamine reduced leucine transport, transamination, and decarboxylation. In contrast, increasing the concentration of oleic acid from 0 to 2 mM dose-dependently stimulated leucine transamination, decarboxylation, and oxidation of carbons 2–6. Oleic acid also enhanced the abundance of cytosolic BCAA transaminase, while reducing the phosphorylated level (inactive state) of the E1α subunit of the mitochondrial branched-chain α-ketoacid dehydrogenase complex. Thus, hypoglycemia or ketosis in early lactation does not likely affect BCAA metabolism in mammary epithelial cells. Increasing circulating levels of BCAA and oleic acid may have great potential to increase the syntheses of glutamate, glutamine, aspartate, alanine, and asparagine by lactating mammary glands, thereby leading to enhanced production of milk for suckling neonates.

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Abbreviations

BCAA:

Branched-chain amino acids

BCAT:

Branched-chain amino acid aminotransferase

BCKA:

Branched-chain α-ketoacids

BCKAD:

Branched-chain α-ketoacid dehydrogenase

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

α-KIC:

α-Ketoisocaproate

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Acknowledgments

Jian Lei was supported by a Postgraduate Scholarship from South China Agricultural University. Work in the authors’ laboratories was supported by National Natural Science Foundation of China grants (#31172217, 30901041), the Thousand-People Talent program at China Agricultural University, Chinese Universities Scientific Fund (2012RC024), National Research Initiative Competitive Grants No. 2008-35206-18764 and 2008-35203-19120 from the USDA National Institute of Food and Agriculture, Texas AgriLife Research Hatch Project no. H-8200, and American Heart Association. We thank Dr. Susan Hutson and Dr. Christopher Lynch for the kind provision of BCAT and BCKAD E1α antibodies, respectively.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. College of Animal Science, South China Agricultural University, Guangzhou, 510642, People’s Republic of China

    Jian Lei, Dingyuan Feng, Yongliang Zhang & Guoyao Wu

  2. Department of Animal Science and Faculty of Nutrition, Texas A&M University, College Station, TX, 77843, USA

    Jian Lei, Sudath Dahanayaka, Xilong Li, Kang Yao, Junjun Wang & Guoyao Wu

  3. State Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, 100193, China

    Junjun Wang, Zhenlong Wu, Zhaolai Dai & Guoyao Wu

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  1. Jian Lei
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  2. Dingyuan Feng
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Correspondence to Jian Lei, Dingyuan Feng or Guoyao Wu.

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Lei, J., Feng, D., Zhang, Y. et al. Regulation of leucine catabolism by metabolic fuels in mammary epithelial cells. Amino Acids 43, 2179–2189 (2012). https://doi.org/10.1007/s00726-012-1302-2

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