| Autor: |
Punetha M; ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India., Chouhan VS; ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India., Sonwane A; ICAR-Indian Veterinary Research Institute, Division of Animal Genetics, Izatnagar, Bareilly, Uttar Pradesh, 243122, India., Singh G; ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India., Bag S; ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India., Green JA; University of Missouri-Columbia, Division of Animal Science, Columbia, Missouri, USA., Whitworth K; University of Missouri-Columbia, Division of Animal Science, Columbia, Missouri, USA., Sarkar M; ICAR-Indian Veterinary Research Institute, Physiology & Climatology Division, Izatnagar, Bareilly, Uttar Pradesh, 243122, India. msarkar24@gmail.com. |
| Abstrakt: |
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. |
