天府肉鸭腔上囊胚胎及胚后生长发育及神经肽表达的研究

【作者】 廖婷彬；

【导师】 方静；

【作者基本信息】 四川农业大学 ， 基础兽医学， 2009， 硕士

【摘要】 本研究运用常规解剖、光镜、透射电镜以及免疫组织化学技术,对天府肉鸭胚胎及胚后生长发育过程中腔上囊的组织形态和亚细胞结构,神经肽的表达进行了动态研究,探讨腔上囊的生长发育规律以及神经肽在其中的形态构筑和生理功能,为进一步研究神经-免疫-内分泌网络提供理论依据,为基础研究和生产实践提供参考。通过不同时期天府肉鸭腔上囊的宏观指标测量和显微结构观察,将腔上囊的胚后发育分为继续发育期(0～8周龄)、持续成熟期(8～14周龄)和退化期(17～32周龄)三个阶段。对宏观指标进行回归分析可知,腔上囊绝对重量和器官指数与时间均有明确的函数关系:胚胎期腔上囊增重呈幂函数关系,器官指数的变化呈二次方程函数;胚后1～14周龄腔上囊绝对重量呈S形曲线增长,1～32周龄器官指数的变化为三次方程函数;胚胎期腔上囊绝对重量与胚体重之间呈三次方程函数关系。26日胚龄的腔上囊组织结构分化发育基本趋于完善;芽突的出现与消失是天府肉鸭腔上囊淋巴滤泡发生的重要阶段;在胚胎和胚后发育过程中,腔上囊淋巴细胞发生凋亡时,细胞核染色质的变化最明显。在天府肉鸭腔上囊的胚胎和胚后发育过程中,利用免疫组化对5种神经肽的表达进行定位研究发现:降钙素基因相关肽(Calcitonin gene-relatedpeptide,CGRP)阳性反应最早出现在20日胚龄腔上囊的黏膜上皮中;在0～14周龄出现于滤泡中;肌层和小动脉平滑肌为阳性反应;17周龄之后CGRP在腔上囊中呈阴性。20日胚龄～32周龄血管活性肠肽(Vasoactive intestinal Peptide,VIP)在滤泡和滤泡相关上皮中呈阳性;除1～14周龄外滤泡间上皮也呈阳性;肌层和小动脉平滑肌呈阳性;5～29周龄固有膜中见少量阳性神经纤维,呈结节状。神经肽Y(Neuropeptide Y,NPY)在滤泡间上皮呈阴性反应;0～29周龄滤泡和滤泡相关上皮呈阳性;20日胚龄～32周龄肌层和小动脉平滑肌呈阳性。生长抑素(Somatostatin,SS)阳性细胞在腔上囊的退化期出现于滤泡间上皮和滤泡髓质中,其胞质中的阳性颗粒较粗大。胃泌素(Gastrin,GAS)阳性反应最早在22日胚龄腔上囊的粘膜上皮中出现;在滤泡相关上皮中持续至26周龄,在滤泡间上皮中持续至3周龄:其在滤泡中持续的时间较短,皮质为1～8周龄,髓质为26同胚龄～1周龄。腔上囊胚胎及胚后发育表现出明显的增龄变化特点。5种神经肽阳性细胞在天府肉鸭的腔上囊中广泛分布,它们首次出现的时间不同,且在胚胎及胚后发育过程中,其数量呈现一定的增龄变化规律。鸭腔上囊是神经系统以外产生CGRP、VIP、NPY、SS、GAS的又一个重要源泉。更多还原

【Abstract】 By the methods of routine anatomic,morphology,transmission electron microscope and immunhistochemistry,the structure of the bursa of fabricius in Tianfu duck and the distribution of five neuropeptides in the bursa were studied during the embryonic and post embryonic development.The developmental pattern,the expression characteristics and physiological function of neuropeptides in the bursa were investigated,which provided theoretical basis for fundamental research,production practice and investigate nervous-immune-endocrine net.The post embryonic development of bursa can be divided into three periods:the continued growth period(0-8weeks),the maturated durative period(8-14weeks) and the degraded period(17-32weeks).The functional relation of the bursal weight and the time were taken on the power function at embryonic stage and the sigmoid curve at 1-14 weeks after hatching.The functional relation of the bursal index and the time were taken on the quadratic function at embryonic stage while the cubic function at 1-32 weeks after hatching.The relationship between the bursal weight and body weight was showed itself to the cubic model at embryonic stage.The histological structure of the bursa was well-developed at 26 days embryonic stage.The emergence and disappearance of the bud break was an important stage during the bursal nodule development.The nuclei of apoptotic lymphocytes were obviously changed during the embryonic and post embryonic development.The positive reaction of calcitonin gene-relatedpeptide(CGRP) originally appeared in the bursal epithelium at 20 days embryonic stage and occurred in follicle at 0-14 weeks after hatching.The smooth muscle layer and arteriolar smooth muscle reacted positively.The bursa showed CGRP negative reaction.Vasoactive intestinal peptide(VIP) positive cells were found in the follicle-associated epithelium(FAE) and follicle from 20 days embryonic stage to 32 weeks after hatching.Interfollicle epithelium(IFE) also showed positive reaction at that time except 1-14 weeks.The smooth muscle layer and arteriolar smooth muscle reacted positively.The VIP positive fibers were located in lamina propria from 5 to 29 weeks after hatching.IFE showed negative reaction of neuropeptide Y(NPY) in each group.FAE and follicle showed NPY positive reaction from 0 to 29 weeks after hatching.The smooth muscle layer and arteriolar smooth muscle reacted positively from 20 days to 32 weeks.Somatostatin(SS) positive cells were observed in the IFE and follicle medulla at the degraded period.The positive reaction particle of brown color was big.Gastrin(GAS) positive cells were detected in FAE from 22 days embryonic stage to 26 weeks after hatching,in IFE from 22 days to 3 weeks.However,the GAS-positive expression of follicle appeared in different time.The bursa of Tianfu duck showed the characteristics of age-related changes during the embryonic and post embryonic development.The positive cells of CGRP,VIP,NPY, SS and GAS were widely distributed in the Tianfu duck bursa at different time,whose number showed the characteristics of age-related changes.The duck bursa was an important source of CGRP,VIP,NPY,SS and GAS,besides the nervous system.更多还原