促性腺激素释放激素(GnRHs)在大菱鲆卵巢不同发育时期的基因表达分析

The expression of GnRHs during ovarian development in turbot (Scophthalmus maximus)

  • 摘要:
    目的 探究大菱鲆促性腺激素释放激素GnRHs在卵巢发育不同时期脑组织中的表达模式和关键亚型定位。
    方法 本实验通过苏木素-伊红(H.E)染色明确其卵巢发育分期与脑区结构;利用实时荧光定量PCR (qRT-PCR)技术检测gnrhs各亚型在不同发育阶段脑区中的mRNA水平;采用原位杂交技术分析关键亚型gnrh1在脑组织中分布定位。
    结果 大菱鲆端脑、中脑、小脑、下丘脑和延脑呈规律性线性排列,且中脑面积占比最大;卵巢发育经历了卵黄生成前期、卵黄生成早期、卵黄生成后期、核迁移期和闭锁期5个阶段。gnrh3在卵黄生成后期的脑组织中显著高表达;而gnrh1与gnrh2则在核迁移期的脑组织中显著上调。同时,在卵巢整个发育过程中,gnrh1在各脑区中的表达水平显著高于其他亚型,并在核迁移期的端脑中呈现特异性高表达。原位杂交结果进一步证实,gnrh1阳性信号富集于腹侧端脑及下丘脑。
    结论 大菱鲆高面积占比的中脑,预示其具备发达的视觉信息处理能力,从而更好适应复杂的底栖环境,同时gnrh1在核迁移期端脑中特异性高表达,影响卵母细胞最后成熟进程,是调控大菱鲆卵巢发育与成熟的关键亚型。系统解析了大菱鲆脑部结构特征,揭示了GnRHs各亚型在卵巢发育过程中的表达规律,为探究神经内分泌调控机制及研发高效生殖调控技术提供了重要理论依据。

     

    Abstract: The turbot (Scophthalmus maximus) is a marine fish of high commercial value and wildly cultured in Europe and China. Gonadotropin-releasing hormones (GnRHs), as a neuropeptide of sex hormone, stimulate synthesis and release of pituitary gonadotropins, inducing vertebrate gonads to provide sex steroids for the formation and maturation of gametes. Hence GnRH has been proven to be the key signal molecule in the hypothalamus-pituitary-gonadal axis. However, their dynamics expression and functional roles during the ovarian developmental in S. maximus remained not fully elucidated. The present study aimed to investigate the expression patterns and key subtype localization of GnRHs in the brain of turbot during ovarian developmental. Ovarian developmental stages and brain structures were identified using hematoxylin and eosin (H.E) staining. The mRNA levels of gnrhs in various brain regions during ovarian developmental were detected by quantitative real-time PCR (qRT-PCR). The structure of S. maximus brain exhibited a regular linear arrangement of the telencephalon, mesencephalon, cerebellum, hypothalamus, and medulla oblongata, with highly differentiated functions and the mesencephalon occupying the largest proportion. Ovarian development obtained the typical pre-vitellogenic, early-vitellogenic, late-vitellogenic, nuclear migration, and atresia stages. The mRNA levels of gnrh3 was peaked at the late-vitellogenic stage, while gnrh1 and gnrh2 were peaked at the nuclear migration stage. Meanwhlie, the mRNA levels of gnrh1 in all brain regions were significantly higher than those of other subtypes, the highest values were observed in the telencephalon at the nuclear migration stage. In situ hybridization further confirmed that gnrh1-positive signals were enriched in the ventral telencephalon and hypothalamus. Thus, the large relative area of the S. maximus mesencephalon confers enhanced visual information processing capacity, enabling adaptation to complex benthic environments. Moreover, the heightened expression of gnrh1 specifically in the telencephalon during the nuclear migration regulates the final maturation of oocytes, identifying it as the key subtype regulating ovarian development and maturation in S. maximus. This study systematically delineates the brain architecture of the S. maximus and uncovers the expression profiles of GnRH subtypes throughout ovarian development. These findings provide a crucial theoretical foundation for investigating neuroendocrine regulatory mechanisms and for the development of efficient reproductive management strategies in captivity.

     

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