Impact of Cell Culture and Copper Dose on Gene Expression in Bovine Liver

• Published in: Biological trace element research Vol. 200; no. 5; pp. 2113 - 2121
• Main Authors: Tillquist, N. M., Thorndyke, M. P., Thomas, T. A., Coleman, S. J., Engle, T. E.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-05-2022; Springer Nature B.V
• Subjects: Abundance; Actin; Adenosine triphosphatase; Aldehyde dehydrogenase; Aldehydes; Animals; Antioxidants; Apolipoprotein A; Betaine-homocysteine S-methyltransferase; Biochemistry; Biomedical and Life Sciences; Biotechnology; Carbonic anhydrase; Carbonic anhydrase II; Cattle; Cell culture; Cell Culture Techniques; Copper; Copper - metabolism; Copper - pharmacology; Cytochrome-c oxidase; Cytochromes; Dehydrogenase; Dehydrogenases; Experiments; Flavin; Flavin reductase; Gene Expression; Genes; Glutamate dehydrogenase; Glutathione; Hepatocytes; Homocysteine; Life Sciences; Liver; Liver - metabolism; Methyltransferase; Nutrition; Oncology; Protein disulfide-isomerase; Reductases; Relative abundance; Superoxide dismutase; Superoxide Dismutase - metabolism; Transportation; Zinc; Beef cattle; Cell culture; Copper homeostasis; Liver; Relative abundance
• ISSN: 0163-4984; 1559-0720
• DOI: 10.1007/s12011-021-02829-5

The objectives of these experiments were to investigate (1) the relative abundance of transcripts for Cu-responsive genes in whole bovine liver vs. cultured hepatocytes and (2) the influence of Cu dose on the relative abundance of transcripts for Cu-responsive genes in cultured bovine hepatocytes. Experiment 1: Liver samples were obtained immediately post-mortem from one healthy Angus steer. Half of the tissue samples were placed in RNA
later
solution; the remaining half was used to isolate hepatocytes. Experiment 2: A subset of cultured hepatocytes was incubated in media containing: 0 mg/L, 0.10 mg/L, 1.0 mg/L, 10.0 mg/L, and 100 mg/L Cu for 1 h. Transcripts analyzed were aldehyde dehydrogenase (ALDH2), apolipoprotein A-1 (APOA1), antioxidant 1 (ATOX1), ATPase copper transporting alpha (ATP7A), ATPase copper transporting beta (ATP7B), betaine homocysteine methyltransferase (BHMT), flavin reductase (BLVRB), carbonic anhydrase II (CA2), copper chaperone for superoxide dismutase (CCS), cytochrome c oxidase copper chaperone (COX17), Cu transporter 1 (CTR1), glutamate dehydrogenase (GLUD1), glutathione synthetase (GSS), protein disulfide isomerase A3 (PDIA3), and superoxide dismutase (Cu–Zn) (SOD1). Β-Actin (ACTB) was selected as the endogenous control in both experiments. Experiment 1: Whole liver had greater (
P
 < 0.01) relative abundance of mRNA for APOA1, ATOX1, ATP7A, ATP7B, COX17, CTR1, ALDH2, BHMT, BLVRB, CA2, GLUD1, and GSS when compared with cultured hepatocytes. Experiment 2: Copper dose impacted all identified transcripts. These results indicate that the relative abundance of Cu-responsive transcripts is different in whole vs. cultured hepatocytes and that the relative abundance of Cu-responsive genes is dependent on Cu dose in cultured hepatocytes.