Acrosome Formation And Components Of Acrosome Complex In Gampsocleis Gratiosa

Orthoptera consists of Caelifera and Ensifera in which Gryloidea and Tettigonioidea are the main researched groups.Collect the common species of Caelifera,Gryloidea and Tettigonioidea and observe the spermatogenesis of them by light microscopy,respectively.The results showed that the heads of sperms point to the free apical part of testis tubular and that the flagella point to the basal part of testis tubular in the spermiogenesis in Caelifera and Grylloidea.Eventually,tails of sperms are firstly released from testis tubules.Opposite to the Caelifera and Gryloidea,the heads of sperms point to the basal part of testis tubular and that the flagella point to the free apical part of testis tubular in the spermiogenesis in Tettigonioidea,the heads of whose sperms are released from testis tubules.The polarization of spermatid cysts among the three groups may be associated with fertilization and the acrosome complex of sperm is the key organelle in the process of fertilization.Gampsocleis gratiosa Brunner von Wattenwyl belongs to Tettigonioidea in Orthoptera.The adults are easy to raise.The large size and the typical structure of the acrosome complex make it to be a useful model to explore the acrosome formation in Orthoptera.The structure and changes of morphology of acrosome complex were studied by TEM(transmission electron microscope),FL(fluorescence labeling)and PAS staining in this thesis;the compositions were demonstrated by comparison ultrastructure with specific staining;the role of microfilament in the acrosome formation was analyzed by the means of treated with cytochalasin D.This study provides the basis for the research on mechanism of acrosome formation in Tettigonioidea and lays the foundation for further exploring the mechanism of spermiogenesis and evolutionary relationship in Orthoptera.Based on the dynamic of the morphology and relative positions of the acrosome and nucleus,acrosome formation in G.gratiosa was divided into seven phases: early round,middle round,late round,rhombic,cylindrical,transforming and mature phases,by paraffin section,frozen section and ultrathin section.Acrosome formation starts with pro-acrosome from Golgi secretion.At the early round and middle round,the pro-acrosome constantly enlarges approaches the nucleus and the pro-acrosome locates the back of nucleus,while the nucleus is condensing;at the late round phase,the size of pro-acrosome is similar to that of nucleus and pro-acrosome rotates by adhering to the nucleus until arriving at the top of nucleus and becomes the acrosome proper;at the rhombic phase,acrosome proper extends along both sides of nucleus and a perforatorium is produced beneath the acrosome proper;at the cylindrical phase,perforatorium also enlarges along both sides of nucleus;eventually,the extra-acrosomal layer is produced on the outside of the acrosome proper,and forms the complete arrow-like acrosome complex.In addition,the PAS staining of the paraffin section demonstrates that polysaccharide exists in the pro-acrosome and acrosome proper from Golgi.In consideration of regular conjugation of polysaccharide and protein in cels,it is proposed that pro-acrosome and acrosome proper are mainly composed of glycoprotein.The results of fluorescence labeling Factin showed the emergence,shape,location and dynamic of microfilament always accompany the perforatorium,so it is speculated that microfilament is one of the elements of the perforatorium.The results from TEM(transmission electron microscope)and FL(fluorescence labeling)showed that microfilament and microtubule play an important role in the whole process of acrosome formation.Microfilament supports the perforatorium as a component of acrosome complex because of the dynamics of microfilament being consistent with that of perforatorium.Microtubule appears on the periphery of nucleus and acrosome complex at the transforming phase,so microtubule may shape the nucleus and acrosome complex.In this research,we further study the role of microfilament in the G.gratiosa by treating with cytochalasin and the results show microfilament is identical in two groups of sperms at the late stage of acrosome formation.But the microfilaments in the treated sperm are less and more incompact than that of normal sperm at the early stage.The results prove from one side that microfilaments play an important role in the process of acrosome formation.By comprehensive analysis of the results,the conclusions are as follows:(1)From the perspective of spermatogenesis,Gryl oidea has a closer evolvement relationship with Caelifera than Tettigonioidea.(2)Based on the dynamic of the morphology and positions of the acrosome and nucleus, acrosome formation in G.gratiosa was divided into seven phases: early round,middle round,late round,rhombic,cylindrical,transforming and mature phases.Taking into consideration the distribution of microfilament and microtubule,we drew a model pattern of acrosome formation in G.gratiosa.(3)In G.gratiosa,the acrosome proper is mainly composed of glycoprotein and microfilament is one of the components of perforatorium.It is speculated that microfilament supports the acrosome complex and that microtubule shapes the nucleus and acrosome complex.