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Peroxisome proliferator-activated receptor β/δ does not regulate glucose uptake and lactose synthesis in bovine mammary epithelial cells cultivated in vitro

Published online by Cambridge University Press:  26 June 2018

Jayant Lohakare ,
Johan S Osorio  and
Massimo Bionaz
Jayant Lohakare
Affiliation:
Department of Animal and Rangeland Sciences, Oregon State University, Corvallis, OR 9733, USA
Johan S Osorio
Affiliation:
Department of Animal and Rangeland Sciences, Oregon State University, Corvallis, OR 9733, USA
Massimo Bionaz*
Affiliation:
Department of Animal and Rangeland Sciences, Oregon State University, Corvallis, OR 9733, USA
*
*For correspondence; e-mail: [email protected]
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Abstract

The hypothesis of the study was that inhibition of PPARβ/δ increases glucose uptake and lactose synthesis in bovine mammary epithelial cells by reducing the expression of the glucose transporter mRNA destabiliser calreticulin. Three experiments were conducted to test the hypothesis using immortalised bovine mammary alveolar (MACT) and primary bovine mammary (PBMC) cells. In Experiment 1, the most effective dose to inhibit PPARβ/δ activity among two synthetic antagonists (GSK-3787 and PT-s58) was assessed using a gene reporter assay. In Experiment 2, the effect on glucose uptake and lactose synthesis was evaluated by measuring glucose and lactose in the media and expression of related key genes upon modulation of PPARβ/δ using GSK-3787, the synthetic PPARβ/δ agonist GW-501516, or a combination of the two in cells cultivated in plastic. In Experiment 3, the same treatments were applied to cells cultivated in Matrigel and glucose and lactose in media were measured. In Experiment 1 it was determined that a significant inhibition of PPARβ/δ in the presence or absence of fetal bovine serum was achieved with ≥ 1000 nm GSK-3787 but no significant inhibition was observed with PT-s58. In Experiment 2, inhibition of PPARβ/δ had no effect on glucose uptake and lactose synthesis but they were both increased by GW-501516 in PBMC. The mRNA abundance of PPARβ/δ target gene pyruvate dehydrogenase kinase 4 was increased but transcription of calreticulin was decreased (only in MACT cells) by GW-501516. Treatment with GSK-3787 did not affect the transcription of measured genes. No effects on glucose uptake or lactose synthesis were detected by modulation of PPARβ/δ activity on cells cultivated in Matrigel. The above data do not provide support for the original hypothesis and suggest that PPARβ/δ does not play a major role in glucose uptake and lactose synthesis in bovine mammary epithelial cells.

Keywords

Peroxisome proliferator-activated receptor β/δmammary epithelial cellscattlelactoseglucose
Type
Research Article
Information
Journal of Dairy Research , Volume 85 , Issue 3 , August 2018 , pp. 295 - 302
Copyright
Copyright © Hannah Dairy Research Foundation 2018 

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Footnotes

†

Present address: Department of Agriculture–Animal Science, University of Arkansas at Pine Bluff, Pine Bluff, AR 71601, USA.

‡

Present address: Department of Dairy and Food Sciences, South Dakota State University, Brookings, SD 57007, USA.

References

Annison, EF & Linzell, JL 1964 The oxidation and utilization of glucose and acetate by the mammary gland of the goat in relation to their over-all metabolism and milk formation. Journal of Physiology 175 372–385Google Scholar
Bionaz, M & Loor, JJ 2007 Identification of reference genes for quantitative real-time PCR in the bovine mammary gland during the lactation cycle. Physiological Genomics 29 312–319Google Scholar
Bionaz, M & Loor, JJ 2011 Gene networks driving bovine mammary protein synthesis during the lactation cycle. Bioinformatics and Biology Insights 5 83–98Google Scholar
Bionaz, M, Periasamy, K, Rodriguez-Zas, SL, Everts, RE, Lewin, HA, Hurley, WL & Loor, JJ 2012 Old and new stories: revelations from functional analysis of the bovine mammary transcriptome during the lactation cycle. PLoS ONE 7 e33268Google Scholar
Bionaz, M, Chen, S, Khan, MJ & Loor, JJ 2013 Functional role of PPARs in ruminants: potential targets for fine-tuning metabolism during growth and lactation. PPAR Research 2013 684159Google Scholar
Bionaz, M, Osorio, J & Loor, JJ 2015 TRIENNIAL LACTATION SYMPOSIUM: nutrigenomics in dairy cows: nutrients, transcription factors, and techniques. Journal of Animal Science 93 5531–5553Google Scholar
Blum, JL, Zeigler, ME & Wicha, MS 1989 Regulation of mammary differentiation by the extracellular matrix. Environmental Health Perspectives 80 71–83Google Scholar
Cengiz, E & Tamborlane, WV 2009 A tale of Two compartments: interstitial versus blood glucose monitoring. Diabetes Technology & Therapeutics 11 S11–S16Google Scholar
Connaughton, S, Chowdhury, F, Attia, RR, Song, S, Zhang, Y, Elam, MB, Cook, GA & Park, EA 2010 Regulation of pyruvate dehydrogenase kinase isoform 4 (PDK4) gene expression by glucocorticoids and insulin. Molecular and Cellular Endocrinology 315 159–167Google Scholar
Harris, RA, Bowker-Kinley, MM, Huang, B & Wu, P 2002 Regulation of the activity of the pyruvate dehydrogenase complex. Advances in Enzyme Regulation 42 249–259Google Scholar
Hillreiner, M, Muller, NI, Koch, HM, Schmautz, C, Kuster, B, Pfaffl, MW & Kliem, H 2017 Establishment of a 3D cell culture model of primary bovine mammary epithelial cells extracted from fresh milk. In Vitro Cellular and Developmental Biology: Animal 53 706–720Google Scholar
Hosseini, A, Sharma, R, Loor, JJ & Bionaz, M 2013 Transcriptomics comparison of Mac-T cells versus mammary tissue during late pregnancy and peak lactation. Advances in Dairy Research 1 1–12Google Scholar
Hu, H, Wang, J, Bu, D, Wei, H, Zhou, L, Li, F & Loor, JJ 2009 In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow. PloS One 4 e7636Google Scholar
Jedrzejczak, M & Szatkowska, I 2014 Bovine mammary epithelial cell cultures for the study of mammary gland functions. In Vitro Cellular and Developmental Biology: Animal 50 389–398Google Scholar
Jeoung, NH & Harris, RA 2010 Role of pyruvate dehydrogenase kinase 4 in regulation of blood glucose levels. Korean Diabetes Journal 34 274–283Google Scholar
Kadegowda, AK, Bionaz, M, Piperova, LS, Erdman, RA & Loor, JJ 2009 Peroxisome proliferator-activated receptor-gamma activation and long-chain fatty acids alter lipogenic gene networks in bovine mammary epithelial cells to various extents. Journal of Dairy Science 92 4276–4289Google Scholar
Lennerz, BS, Alsop, DC, Holsen, LM, Stern, E, Rojas, R, Ebbeling, CB, Goldstein, JM & Ludwig, DS 2013 Effects of dietary glycemic index on brain regions related to reward and craving in men. American Journal of Clinical Nutrition 98 641–647Google Scholar
Lin, Y, Sun, X, Hou, X, Qu, B, Gao, X & Li, Q 2016 Effects of glucose on lactose synthesis in mammary epithelial cells from dairy cow. BMC Veterinary Research 12 81Google Scholar
Naruhn, S, Toth, PM, Adhikary, T, Kaddatz, K, Pape, V, Dorr, S, Klebe, G, Muller-Brusselbach, S, Diederich, WE & Muller, R 2011 High-affinity peroxisome proliferator-activated receptor beta/delta-specific ligands with pure antagonistic or inverse agonistic properties. Molecular Pharmacology 80 828–838Google Scholar
Nikkhah, A, Furedi, CJ, Kennedy, AD, Crow, GH & Plaizier, JC 2008 Effects of feed delivery time on feed intake, milk production, and blood metabolites of dairy cows. Journal of Dairy Science 91 4249–4260Google Scholar
Oliver, WR Jr., Shenk, JL, Snaith, MR, Russell, CS, Plunket, KD, Bodkin, NL, Lewis, MC, Winegar, DA, Sznaidman, ML, Lambert, MH, Xu, HE, Sternbach, DD, Kliewer, SA, Hansen, BC & Willson, TM 2001 A selective peroxisome proliferator-activated receptor delta agonist promotes reverse cholesterol transport. Proceedings of the National Academy of Sciences of the United States of America 98 5306–5311Google Scholar
Ordelheide, AM, Heni, M, Thamer, C, Machicao, F, Fritsche, A, Stefan, N, Haring, HU & Staiger, H 2011 In vitro responsiveness of human muscle cell peroxisome proliferator-activated receptor delta reflects donors’ insulin sensitivity in vivo. European Journal of Clinical Investigation 41 1323–1329Google Scholar
Osorio, JS & Bionaz, M 2017 Plasmid transfection in bovine cells: optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay. Gene 626 200–208Google Scholar
Palkar, PS, Borland, MG, Naruhn, S, Ferry, CH, Lee, C, Sk, UH, Sharma, AK, Amin, S, Murray, IA, Anderson, CR, Perdew, GH, Gonzalez, FJ, Muller, R & Peters, JM 2010 Cellular and pharmacological selectivity of the peroxisome proliferator-activated receptor-beta/delta antagonist GSK3787. Molecular Pharmacology 78 419–430Google Scholar
Riahi, Y, Sin-Malia, Y, Cohen, G, Alpert, E, Gruzman, A, Eckel, J, Staels, B, Guichardant, M & Sasson, S 2010 The natural protective mechanism against hyperglycemia in vascular endothelial cells: roles of the lipid peroxidation product 4-hydroxydodecadienal and peroxisome proliferator-activated receptor delta. Diabetes 59 808–818Google Scholar
Rosa, F, Osorio, JS, Trevisi, E, Yanqui-Rivera, F, Estill, CT & Bionaz, M 2017 2,4-Thiazolidinedione treatment improves the innate immune response in dairy goats with induced subclinical mastitis. PPAR Research 2017 7097450Google Scholar
Ruijter, JM, Ramakers, C, Hoogaars, WM, Karlen, Y, Bakker, O, van den Hoff, MJ & Moorman, AF 2009 Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Research 37 e45Google Scholar
Sasaki, M & Keenan, TW 1978 Membranes of mammary-gland .18. 2-deoxy-D-glucose and 5-Thio-D-glucose decrease lactose content, inhibit secretory maturation and depress protein-synthesis and secretion in lactating Rat mammary-gland. International Journal of Biochemistry 9 579Google Scholar
Song, S, Attia, RR, Connaughton, S, Niesen, MI, Ness, GC, Elam, MB, Hori, RT, Cook, GA & Park, EA 2010 Peroxisome proliferator activated receptor alpha (PPARalpha) and PPAR gamma coactivator (PGC-1alpha) induce carnitine palmitoyltransferase IA (CPT-1A) via independent gene elements. Molecular and Cellular Endocrinology 325 54–63Google Scholar
Totary-Jain, H, Naveh-Many, T, Riahi, Y, Kaiser, N, Eckel, J & Sasson, S 2005 Calreticulin destabilizes glucose transporter-1 mRNA in vascular endothelial and smooth muscle cells under high-glucose conditions. Circulation Research 97 1001–1008Google Scholar
Vandesompele, J, De Preter, K, Pattyn, F, Poppe, B, Van Roy, N, De Paepe, A & Speleman, F 2002 Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology 3 RESEARCH0034Google Scholar
Wang, YF, Chao, HR, Wu, CH, Tseng, CH, Kuo, YT & Tsou, TC 2010 A recombinant peroxisome proliferator response element-driven luciferase assay for evaluation of potential environmental obesogens. Biotechnology Letters 32 1789–1796Google Scholar
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