克氏原螯虾眼柄典型神经内分泌胞团调控卵巢发育功能的介入干扰研究

Revealing the in vivo secretory and remote controlling/regulation function of the neuroendocrine cells in intact-neural-tissue is very important to clarify the accurate structure and regulation function of the postulated crustacean neuroendocrine center named XO-SG. Addressing this challenge, our study using Procambrus clarkii as specimen developed an eyestalk intervention method to explore the regulation function of the XO-SG and related structure basing on following steps: (a) obstain the high-resolution global perspective situation information of the suspect neuroendocrine cells in the eyestalks, including their nerve fiber orientation and synaptic connections with the technology of eyestalk-brain ganglia intact-tissue CLARITY, combined with serial eyestalk HE slices and 3DHISTEC scanning; (b) disturb suspected eyestalk neuroendocrine cell metabolism and secretion via intervention in vicinity with cell membrane permeability enchancer, nerurotransmitter receptor antagonist/ activator, the depletion agent of ATP, and related ions; (c) comparative analysis, among eyestalk intervention groups,eyestalk ablation group and blank control group, of gonad promotion and molting induction effect, as well as level changes of Vg etc. in the heamolymph, to uncover the reproduction regulation function of XO-SG of P. clarkii and its related tissue specificity in its eyestalk. This study will update our knowledge about the structure and function of XO-SG by clarifing the relationship between the typical ovarian development controlling function and some specific tissue or cells in the eyestalks. Meanwhile, the intervention technique used in this study will be an effective method for the structure-functional analysis of XO-SG in vivo , in crustaceans. Then the foreground application of eyestalk intervention in rgulating ovarian development will be discussed.

从整体层面和活体水平揭示眼柄神经内分泌胞团的自身分泌和远端调控功能，对于阐明XO-SG的准确结构与眼柄切除的生殖调控机制至关重要。本研究以克氏原螯虾为样本，（1）采用眼柄-脑组织整体CLARITY技术，结合眼柄连续HE切片和3DHISTEC扫描，查明眼柄神经内分泌细胞聚集、走向和突触连接等详细空间结构信息；（2）在眼柄嫌疑胞团附近介入缓释性的细胞膜透性增强剂、神经递质受体拮抗剂和激活剂、ATP耗竭剂、关键离子等，干扰眼柄神经内分泌细胞的代谢或分泌；（3）比较分析介入处理组关键生理指标和性腺发育速度与眼柄切除组及空白对照组异同，从活体水平和“XO-SG-脑”整体层面，揭示XO-SG的各部分结构与卵巢发育过程中典型调控功能间的精确关系。研究将补充、更新甲壳动物XO-SG结构与神经内分泌功能知识、建立一种更为直观精确的“结构-功能”活体分析方法，并探索眼柄介入技术在卵巢发育调控中的应用前景。