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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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| Published in: | Scientific reports 2020-04, Vol.10 (1), p.6849-6849, Article 6849 |
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| creator | Punetha, Meeti Chouhan, Vikrant S Sonwane, Arvind Singh, Gyanendra Bag, Sadhan Green, Jonathan A Whitworth, Kristin Sarkar, Mihir |
| description | 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. |
| doi_str_mv | 10.1038/s41598-020-63804-z |
| format | article |
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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.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-020-63804-z</identifier><identifier>PMID: 32321973</identifier><language>eng</language><publisher>England: Nature Publishing Group</publisher><subject>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</subject><ispartof>Scientific reports, 2020-04, Vol.10 (1), p.6849-6849, Article 6849</ispartof><rights>The Author(s) 2020. 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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.</description><subject>Angiogenesis</subject><subject>Animals</subject><subject>Bubalus bubalis</subject><subject>Buffaloes - genetics</subject><subject>Buffaloes - metabolism</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Cell proliferation</subject><subject>Corpus luteum</subject><subject>Corpus Luteum - metabolism</subject><subject>CRISPR</subject><subject>CRISPR-Cas Systems</subject><subject>EGR-1 protein</subject><subject>Egr-2 protein</subject><subject>Egr-3 protein</subject><subject>EGR-4 protein</subject><subject>Female</subject><subject>Fibroblast growth factor 2</subject><subject>Fibroblast Growth Factor 2 - biosynthesis</subject><subject>Fibroblast Growth Factor 2 - genetics</subject><subject>Gene Editing</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Gene Knockout Techniques</subject><subject>Genes</subject><subject>Genome editing</subject><subject>Growth factors</subject><subject>Steroidogenesis</subject><subject>Vascular endothelial growth factor</subject><subject>Vascular Endothelial Growth Factor A - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Punetha, Meeti</au><au>Chouhan, Vikrant S</au><au>Sonwane, Arvind</au><au>Singh, Gyanendra</au><au>Bag, Sadhan</au><au>Green, Jonathan A</au><au>Whitworth, Kristin</au><au>Sarkar, Mihir</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Early growth response gene mediates in VEGF and FGF signaling as dissected by CRISPR in corpus luteum of water buffalo</atitle><jtitle>Scientific reports</jtitle><addtitle>Sci Rep</addtitle><date>2020-04-22</date><risdate>2020</risdate><volume>10</volume><issue>1</issue><spage>6849</spage><epage>6849</epage><pages>6849-6849</pages><artnum>6849</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>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.</abstract><cop>England</cop><pub>Nature Publishing Group</pub><pmid>32321973</pmid><doi>10.1038/s41598-020-63804-z</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0002-4252-4027</orcidid><oa>free_for_read</oa></addata></record> |
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| 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 |
| title | Early growth response gene mediates in VEGF and FGF signaling as dissected by CRISPR in corpus luteum of water buffalo |
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