Key Laboratory of Agricultural - Animal Genetics, Breeding, and Reproduction, Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, People's Republic of China.
Department of Animal Science, College of Agricultural Sciences, Bule Hora University, Bule Hora, Ethiopia.
Anim Biotechnol. 2023 Dec;34(7):2285-2294. doi: 10.1080/10495398.2022.2087665. Epub 2022 Jun 17.
To evaluate the efficacy of oral immunization with active kisspeptin DNA vaccine on the expression of hormone receptor mRNA. For this study, ten 56-day-old Hu breed ram lambs were randomly assigned to the treatment and control groups ( = 5). Treatment Experimental group received C500/pKS-asd and the control group received C500/pVAX-asd (aspartate-β semialdehyde dehydrogenase orally on days 0, 28, and 56, and blood samples were taken at each immunization interval (14-day) and tissues samples were collected at the end of the experimental period (day 98). The collected samples were stored in the refrigerator at -20 °C and liquid nitrogen, respectively, for laboratory examination. Total RNA was extracted from samples using TRIzol reagent and quantitative real-time polymerase chain reaction (QPCR) was used to quantify the levels of KISS1, G protein-coupled receptor-54 (Kiss1r), and gonadotrophin-releasing hormone (GnRH) mRNA in the hypothalamus. Levels of luteinizing hormone receptor (LHR) and luteinizing hormone beta (LHβ) mRNA, and follicle-stimulating hormone receptor (FSHR) and follicle-stimulating hormone beta (FSHβ) mRNA in the testes and pituitary were analyzed, respectively. Further, gonadotropin-releasing hormone receptor (GnRHR) mRNA expression level in the pituitary was measured. Moreover, the Kiss1r concentration level in the blood was measured using an indirect ELISA. The concentration of Kiss1r in the blood was lower in the treatment group than in the control group ( < 0.05). The levels of testicular FSHR and LHR mRNA were significantly lower in the treatment group ( < 0.05) when compared to the control group. Furthermore, the treatment group's levels of hypothalamic KISS1, Kiss1r, and GnRH mRNA were significantly lower ( < 0.05) than the controls. LH, FSH, and GnRHR mRNA expression in the pituitary were also significantly lower in the treatment group ( < 0.01 and < 0.05, respectively). These findings imply that oral immunization with active kisspeptin DNA vaccine suppresses hormone receptor mRNA expression in the ram lambs.
为了评估主动 Kisspeptin DNA 疫苗口服免疫对激素受体 mRNA 表达的疗效。在这项研究中，将 10 只 56 日龄湖羊羔羊随机分为治疗组和对照组（n=5）。治疗实验组接受 C500/pKS-asd，对照组接受 C500/pVAX-asd（天 0、28 和 56 时β-半醛脱氢酶天），并在每次免疫间隔（14 天）时采集血液样本，并在实验结束时（第 98 天）收集组织样本。收集的样本分别储存在冰箱（-20°C）和液氮中，用于实验室检查。使用 TRIzol 试剂从样品中提取总 RNA，并使用定量实时聚合酶链反应（QPCR）定量测量下丘脑 KISS1、G 蛋白偶联受体-54（Kiss1r）和促性腺激素释放激素（GnRH）mRNA 的水平。分析睾丸和垂体中黄体生成素受体（LHR）和黄体生成素β（LHβ）mRNA、卵泡刺激素受体（FSHR）和卵泡刺激素β（FSHβ）mRNA 的水平，分别为。此外，测量垂体中的促性腺激素释放激素受体（GnRHR）mRNA 表达水平。此外，使用间接 ELISA 测量血液中的 Kiss1r 浓度水平。与对照组相比，治疗组血液中的 Kiss1r 浓度较低（<0.05）。与对照组相比，治疗组睾丸 FSHR 和 LHR mRNA 水平显著降低（<0.05）。此外，治疗组下丘脑 KISS1、Kiss1r 和 GnRH mRNA 水平也明显低于对照组（<0.05）。垂体中 LH、FSH 和 GnRHR mRNA 的表达也明显降低（分别为<0.01 和<0.05）。这些发现表明，主动 Kisspeptin DNA 疫苗口服免疫可抑制公羊羔羊的激素受体 mRNA 表达。
