Multifactorial regulation of reproduction in fish

我研究的长期目标是了解垂体促性腺激素和局部衍生的激素和蛋白质生长因子调节鱼类卵巢功能的相互作用机制。我们的目标是描述卵巢信号分子和信号转导途径，这些信号分子和转导途径介导类固醇激素的生成、卵母细胞的最终成熟和排卵，并在整个动物水平上整合这些反应对产卵成功的影响。具体地说，我们正在通过研究促性腺激素、性激素、前列腺素和胰岛素样生长因子对卵泡细胞成熟能力、卵母细胞成熟、卵巢卵泡水化和排卵的交互作用，来研究排卵前期卵巢发育的激素调节。我们还在研究雄性斑马鱼信息素信号的性质，这些信号有助于雌性斑马鱼的内分泌变化和产卵成功。我们的研究调查了卵巢激素及其受体和基因产物的表达对产卵的影响，直到生殖成功。本研究包括卵巢卵泡的体外培养和体内研究，以确定卵巢卵泡成熟和排卵的激素调节。我们常规使用RIA和EIA方法来定量卵巢类固醇激素和PG水平，RT-qPCR和原位杂交方法来表征基因表达，免疫组织化学方法来表征卵巢蛋白激素和受体的分布。我们将利用药理学方法和反义吗啉寡核苷酸进行功能丧失研究，以探讨类固醇激素、前列腺素、多肽及其受体在发育能力和卵泡存活中所起的作用。这一知识是了解生殖适合性的基础，并提供了能够检查内分泌干扰物和毒物影响野生和养殖鱼类卵巢生理的机制的方法。

The long-term objective of my research is to understand the interactive mechanisms by which pituitary gonadotropins and locally derived hormones and protein growth factors regulate ovarian function in fish. The goal is to characterize ovarian signaling molecules and transduction pathways that mediate steroidogenesis, oocyte final maturation and ovulation and to integrate these responses at the whole animal level in terms of effects on spawning success. Specifically, we are studying the hormonal regulation of ovarian development in the preovulatory period through investigations of the interactive effects of gonadotropins, sex steroids, prostaglandins and insulin-like growth factors on the acquisition of follicle cell maturational competence, oocyte maturation, ovarian follicle hydration and ovulation. We are also studying the nature of the pheromonal signals from male zebrafish that contribute to endocrine changes and spawning success in the female. Our studies investigate the expression of ovarian hormones, their receptors and gene products through to reproductive success in terms of effects on spawning. This research involves both in vitro cultures of ovarian follicles and in vivo studies to characterize the hormonal regulation of the maturation and ovulation of ovarian follicles. We routinely use RIA and EIA methods to quantify ovarian steroid hormone and PG levels, RT-qPCR and in situ hybridization to characterize gene expression and immunohistochemical approaches to characterize ovarian protein hormone and receptor distribution. We will use pharmacological approaches and antisense morpholino oligonucleotides for loss of function studies to investigate the roles played by steroid hormones, PGs, peptide hormones and their receptors in relation to developmental competency and follicle survival. This knowledge is fundamental to understanding reproductive fitness and provides methods that enable examination of the mechanisms by which endocrine disrupting compounds and toxicants affect ovarian physiology in both wild and cultured fish.