Expression and localization of fibroblast growth factor (FGF) family in corpus luteum during different stages of estrous cycle and synergistic role of FGF2 and vascular endothelial growth factor (VEGF) on steroidogenesis, angiogenesis and survivability of cultured buffalo luteal cells

Q1 Agricultural and Biological Sciences

Agri Gene Pub Date : 2016-08-01 DOI:10.1016/j.aggene.2016.07.001

S.R. Mishra , M.S. Parmar , V.S. Chouhan , G. Rajesh , V.P. Yadav , M.K. Bharti , Jaya Bharati , T. Mondal , R. Reshma , A. Paul , S.S. Dangi , B.C. Das , L.A. González , G.T. Sharma , G. Singh , M. Sarkar

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引用次数: 10

Abstract

The aim of this study was to document the expression and localization of fibroblast growth factor (FGF) family members comprising of fibroblast growth factor (FGF1, FGF2, FGF7, FGF10), and their receptors (FGFR1, FGFR2, FGFR3, FGFR4, FGFR2IIIB, FGFR2IIIC) in buffalo corpus luteum (CL) obtained at different stages of the estrous cycle. In addition, the synergistic role of FGF2 and/or vascular endothelial growth factor (VEGF) on P₄ secretion and mRNA expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 (CYP11A1), 3-beta-hydroxysteroid dehydrogenase (3βHSD), proliferating cell nuclear antigen (PCNA), BCL-2 associated X protein (BAX) and von willebrand factor (vWF) were studied in luteal cell culture obtained from mid-luteal phase (MLP) of estrous cycle in buffalo. Real-time PCR (qPCR), western blot, and immunohistochemistry were used to investigate mRNA and protein expressions, and the localization of examined factors whereas P₄ secretion was assessed by RIA. The mRNA and protein expression of FGF1 and FGFR1 were maximum (P < 0.05) during MLP whereas FGF2 was maximum (P < 0.05) during early luteal phase (ELP). FGF7, FGF10, FGFR2, FGFR3, FGFR4, FGFR2IIIb, and FGFR2IIIc mRNA and protein expression did not change among luteal phases. FGF family members were localized in cytoplasm of luteal cells as well as in endothelial cells. P₄ secretion in luteal cells treated with FGF2 or VEGF alone showed the maximum values (P < 0.05) with the highest dose at 72 h. P₄ secretion was found to be greater (P < 0.05) in luteal cells treated with FGF2 + VEGF compared to FGF2 or at 72 h of incubation. The mRNA expression of all factors were maximum (P < 0.05) whereas BAX was minimum (P < 0.05) at highest dose cultured for 72 h of luteal cells subjected with either protein alone or in combination. Summarizing, the present findings explore the synergistic role of FGF2 and VEGF on steroidogenesis, angiogenesis, cell viability through an autocrine and paracrine actions in buffalo CL.

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成纤维细胞生长因子(FGF)家族在黄体不同发情周期的表达和定位及FGF2和血管内皮生长因子(VEGF)对培养水牛黄体细胞甾体生成、血管生成和存活能力的协同作用

本研究的目的是记录成纤维细胞生长因子(FGF)家族成员(FGF1, FGF2, FGF7, FGF10)及其受体(FGFR1, FGFR2, FGFR3, FGFR4, FGFR2IIIB, FGFR2IIIC)在发情周期不同阶段获得的水牛黄体(CL)中的表达和定位。此外，本实验还研究了FGF2和/或血管内皮生长因子(VEGF)对水牛黄体发情中期黄体细胞P4分泌和甾体急性调节蛋白(StAR)、细胞色素P450 (CYP11A1)、3- β -羟基类固醇脱氢酶(3βHSD)、增殖细胞核抗原(PCNA)、BCL-2相关X蛋白(BAX)和血管性血维病因子(vWF) mRNA表达的协同作用。采用实时荧光定量PCR (Real-time PCR, qPCR)、免疫印迹(western blot)和免疫组织化学(immunohistochemistry)检测mRNA和蛋白的表达，以及检测因子的定位，同时采用RIA检测P4的分泌。FGF1和FGFR1的mRNA和蛋白表达量最大(P <0.05)，而FGF2最大(P <黄体早期(ELP)。FGF7、FGF10、FGFR2、FGFR3、FGFR4、FGFR2IIIb和FGFR2IIIc的mRNA和蛋白表达在黄体期之间没有变化。FGF家族成员定位于黄体细胞和内皮细胞的细胞质中。FGF2或VEGF单独处理的黄体细胞中P4分泌量最大(P <0.05)， 72 h时剂量最大。P4分泌量更大(P <与FGF2或孵育72 h时相比，FGF2 + VEGF处理的黄体细胞的差异(0.05)。各因子mRNA表达量均最大(P <0.05)， BAX最小(P <单独或联合作用72h的黄体细胞，最高剂量0.05)。综上所述，本研究探讨了FGF2和VEGF通过自分泌和旁分泌作用在水牛CL中对类固醇生成、血管生成和细胞活力的协同作用。

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Agri Gene Agricultural and Biological Sciences-Agricultural and Biological Sciences (miscellaneous)

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期刊介绍： Agri Gene publishes papers that focus on the regulation, expression, function and evolution of genes in crop plants, farm animals, and agriculturally important insects and microorganisms. Agri Gene strives to be a diverse journal and topics in multiple fields will be considered for publication so long as their main focus is on agriculturally important organisms (plants, animals, insects, or microorganisms). Although not limited to the following, some examples of potential topics include: Gene discovery and characterization. Genetic markers to guide traditional breeding. Genetic effects of transposable elements. Evolutionary genetics, molecular evolution, population genetics, and phylogenetics. Profiling of gene expression and genetic variation. Biotechnology and crop or livestock improvement. Genetic improvement of biological control microorganisms. Genetic control of secondary metabolic pathways and metabolic enzymes of crop pathogens. Transcription analysis of beneficial or pest insect developmental stages Agri Gene encourages submission of novel manuscripts that present a reasonable level of analysis, functional relevance and/or mechanistic insight. Agri Gene also welcomes papers that have predominantly a descriptive component but improve the essential basis of knowledge for subsequent functional studies, or which provide important confirmation of recently published discoveries provided that the information is new.