Rapid Communication: Period2 gene silencing increases the synthesis of α^sub s^-casein protein in bovine mammary epithelial cells 1

Period2 (PER2), a core clock gene, encodes a circadian rhythm protein which has been shown to control mammary metabolism in rodents. Whether PER2 regulates milk component synthesis such as αs-casein protein in bovine mammary cells is unknown. Thus, we used gene silencing technology to determine if P...

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Bibliographic Details
Published inJournal of animal science Vol. 95; no. 10; pp. 4510 - 4513
Main Authors Hu, L Y, Wang, M Z, Ouyang, J L, Li, P F, Loor, J J
Format Journal Article
LanguageEnglish
Published Champaign Oxford University Press 01.10.2017
Subjects
Animal lactation
Casein
Cattle
Cell proliferation
Circadian rhythms
Clock gene
Enzyme-linked immunosorbent assay
Epithelial cells
Gene silencing
Genes
Lactation
Mammary gland
Metabolism
Milk
Oligonucleotides
Period 2 protein
Protein biosynthesis
Protein synthesis
Proteins
Ribonucleic acid
RNA
Rodents
siRNA
Transcription
Transfection
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Summary:Period2 (PER2), a core clock gene, encodes a circadian rhythm protein which has been shown to control mammary metabolism in rodents. Whether PER2 regulates milk component synthesis such as αs-casein protein in bovine mammary cells is unknown. Thus, we used gene silencing technology to determine if PER2 silencing could affect αs-casein synthesis and cell growth in cultured primary bovine mammary epithelial cells (BMEC). The BMEC were established by enzymatic digestion of mammary tissue from mid-lactation cows. A transient-transfection technique was used to insert a small interfering RNA (siRNA) oligonucleotide specific for PER2 to inhibit transcription. Control and siRNA-transfected cells were cultured for 48 h. qRT-PCR and ELISA analysis showed that PER2 silencing enhanced the synthesis of 2 kinds of αs-casein (P < 0.05) through upregulating the mRNA level of CSN1S1 and CSN1S2 (P < 0.01). Furthermore, the 3-(4, 5-dimethyl-2-thiazolyl)- 2, 5-diphenyl-2-H-tetrazolium bromide (MTT) results demonstrated that cell proliferation was not affected (P > 0.05). These data led us to hypothesize that PER2 protein may potentially play an important role in the control of milk protein synthesis and, hence, represents a target that can be used to regulate protein synthesis rate during lactation.
ISSN:0021-8812
1525-3163
DOI:10.2527/jas2017.1938