ΔFosB regulates Ca²⁺ release and proliferation of goat mammary epithelial cells.
ΔFosB is a member of the family of transcription factor activating proteins-1 (AP-1) and is known to play important roles in Ca(2+) metabolism processes of osteoblast formation and differentiation in humans and rodents. The postpartum mammary gland is one of the significant organs for Ca(2+) metabolism processes. However, very little information is available on the role of ΔFosB in goat mammary gland. In this investigation, the full-length cDNA of ΔFosB from Xinong Saanen dairy goats was cloned, which contains an open-reading frame (ORF) of 723 bp encoding 240 amino acids. The amino acid sequence is highly homologous with cattle (99.17%). Quantitative real time PCR (QRT-PCR) and western blotting assays showed that ΔFosB was expressed in goat heart, liver, lung, and breast, but little in the hypophysis and spleen. The fluorescence signals revealed that the Ca(2+) was decreased in goat mammary epithelial cells (GMECs) over-expressed ΔFosB at 72h. Consistently, intracellular Ca(2+) was increased in GMECs suppressing expressed ΔFosB at 72 h. QRT-PCR assay showed that ΔFosB positively regulated the mRNA expression of runt related transcription factor 2 (Runx2), SMAD family member 4 (Smad4), S100 calcium binding protein A4 (S100A4) and S100 calcium binding protein A13 (S100A13) genes in GMECs, which had been proven to be relative to calcium metabolism in humans and rodents. Ca(2+) could induce a dose-dependent increase of the ΔFosB mRNA expression and a dose-dependent decrease in cell viability when the GMECs were treated with CaCl2. Suppressing ΔFosB expression in GMECs also inhibited the cell viability. These discoveries suggest that ΔFosB plays important roles in regulating Ca(2+) release and proliferation of the GMECs, which may prove useful in regulation of milk production.
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China. Electronic address: zhenghuiling@nwsuaf.edu.cn.
Huiling
H
Zheng
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
Hui
H
Li
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
Lihui
L
Li
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
Shaoyang
S
Ma
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
Xuemei
X
Liu
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006041:Goats / Q000145:classification / Q000235:genetics; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D016760:Proto-Oncogene Proteins c-fos / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism
D018076:DNA, Complementary; D016760:Proto-Oncogene Proteins c-fos; D018808:Transcription Factor AP-1; D002118:Calcium
Calcium metabolism; Cell viability; Goat; ΔFosB