Early growth response gene mediates in VEGF and FGF signaling as dissected by CRISPR in corpus luteum of water buffalo

The EGR family comprises of EGR 1, EGR 2, EGR 3 and EGR 4 which are involved in the transactivation of several genes. A broad range of extracellular stimuli by growth factors is capable of activating EGR mediated transactivation of genes involved in angiogenesis and cell proliferation. However, thei...

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Bibliographic Details
Published in:Scientific reports 2020-04, Vol.10 (1), p.6849-6849, Article 6849
Main Authors: Punetha, Meeti, Chouhan, Vikrant S, Sonwane, Arvind, Singh, Gyanendra, Bag, Sadhan, Green, Jonathan A, Whitworth, Kristin, Sarkar, Mihir
Format: Article
Language:English
Subjects:
Angiogenesis
Animals
Bubalus bubalis
Buffaloes - genetics
Buffaloes - metabolism
Cell culture
Cell growth
Cell proliferation
Corpus luteum
Corpus Luteum - metabolism
CRISPR
CRISPR-Cas Systems
EGR-1 protein
Egr-2 protein
Egr-3 protein
EGR-4 protein
Female
Fibroblast growth factor 2
Fibroblast Growth Factor 2 - biosynthesis
Fibroblast Growth Factor 2 - genetics
Gene Editing
Gene expression
Gene Expression Regulation
Gene Knockout Techniques
Genes
Genome editing
Growth factors
Steroidogenesis
Vascular endothelial growth factor
Vascular Endothelial Growth Factor A - biosynthesis
Vascular Endothelial Growth Factor A - genetics
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Summary:The EGR family comprises of EGR 1, EGR 2, EGR 3 and EGR 4 which are involved in the transactivation of several genes. A broad range of extracellular stimuli by growth factors is capable of activating EGR mediated transactivation of genes involved in angiogenesis and cell proliferation. However, their role in controlling VEGF A and FGF 2 signaling in the CL of water buffalo is not known. The present study was conducted to understand the role of EGR mediated regulation of VEGF A and FGF 2 signaling in buffalo luteal cells. Towards this goal, luteal cells were cultured and treated with VEGF A and FGF 2 and the mRNA expression pattern of EGR family members were documented. The EGR 1 message was found to be up-regulated in luteal cells of buffalo at 72 hours of culture. The functional validation of EGR 1 gene was accomplished by knocking out (KO) of EGR 1 in cultured luteal cells by CRISPR/Cas9 mediated gene editing technology. The EGR 1 KO cells were then cultured and stimulated with VEGF A and FGF 2. It was observed that VEGF A and FGF 2 induced angiogenesis, cell proliferation and steroidogenesis in wild type luteal cells, whereas the response of the growth factors was attenuated in the EGR 1 KO cells. Taken together our study provides evidence convincingly that both VEGF and FGF mediate their biological action through a common intermediate, EGR 1, to regulate corpus luteum function of buffalo.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-63804-z