Faculty of Marine Sciences, Ruppin Academic Center, Michmoret, Israel.
Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi, India.
Fish Physiol Biochem. 2023 Oct;49(5):911-923. doi: 10.1007/s10695-023-01230-4. Epub 2023 Aug 7.
The two gonadotropins, FSH and LH, stimulate growth and development of the gonads through gonadal biosynthesis of steroid hormones and growth factors. To date, cDNA sequences encoding gonadotropin subunits have been isolated and characterized from a large number of fish species. Recently, we successfully cloned and characterized gonadotropins (LHβ, FSHβ, and GPα) from the pituitary glands of the catfish, Heteropneustes fossilis. In the present study, we describe herein the production of recombinant stinging catfish, H. fossilis (hf) FSH (rhfFSH) and LH (rhfLH) using the methylotrophic yeast P. pastoris expression system. We further explored the hypothesis that the recombinant gonadotropins can modulate the hypothalamus-pituitary-ovarian (HPO) axis genes (avt, it, gnrh2, kiss2, and cyp19a1a) and regulate their transcriptional profile and steroid levels in relation to their annual developmental stage during preparatory and pre-spawning phases under in-vitro conditions. We found that the different concentrations of recombinant rhfFSH and rhfLH significantly stimulated E2 levels in the preparatory and prespawning season, and also upregulated gonadal aromatase gene expression in a dose dependent manner. Our results demonstrate that the yeast expression system produced biologically active recombinant catfish gonadotropins, enabling the study of their function in the catfish.
两种促性腺激素,FSH 和 LH,通过性腺类固醇激素和生长因子的生物合成刺激性腺的生长和发育。迄今为止,已经从大量鱼类物种中分离和鉴定了编码促性腺激素亚基的 cDNA 序列。最近,我们成功地从鲶鱼(Heteropneustes fossilis)的垂体中克隆和鉴定了促性腺激素(LHβ、FSHβ 和 GPα)。在本研究中,我们描述了使用甲基营养酵母 P. pastoris 表达系统生产重组鲶鱼,H. fossilis(hf)FSH(rhfFSH)和 LH(rhfLH)。我们进一步探讨了这样一种假设,即重组促性腺激素可以调节下丘脑-垂体-卵巢(HPO)轴基因(avt、it、gnrh2、kiss2 和 cyp19a1a),并调节它们的转录谱和类固醇水平与它们在预备和产卵前阶段的年度发育阶段有关,在体外条件下。我们发现,不同浓度的重组 rhfFSH 和 rhfLH 显著刺激了预备和产卵季节的 E2 水平,并以剂量依赖的方式上调了性腺芳香酶基因的表达。我们的结果表明,酵母表达系统产生了具有生物活性的重组鲶鱼促性腺激素,使我们能够研究它们在鲶鱼中的功能。