| Spermatogenesis is a special and interesting process. A primordial germ cell must endure mitosis, meiosis and spermiogenesis to become a mature sperm. Here we utilized 2-dimensional electrophoresis MALDI-TOF/TOF technology to construct a comparative proteome profile for the mouse first wave of spermatogenesis. Cluster analysis was then performed to better characterize the more specific and unique expression patterns of these different proteins and the distinct expression patterns may decided by their functions during spermatogenesis. In these different proteins, many have been reported associated with spermatogenesis, and their their functions can coincide with their expression modes in our profile, which proves our successful construction.Lmna encoded two isoforms named lamin A/C in mouse testis, they highly expressed in two stages during the first wave of spermatogenesis. Previous studies showed that lamin A/C located in the somatic cells and early germ cells, and male mouse were sterility in lmna KO mouse for the disturbance of meiosis[1]. These location and function were consisted with it's high expression at the 0day,1days time point. However, the second wave of lamin A/C expression occurred in 28days suggests that lamin A/C may involve in spermiogenesis. By western blot we know that lamin A/C expressed both in testis and sperm, immunofluorescence showed that lamin A/C located in acroplaxome and may involved in the mouse spermiogenesis, which we launched a further study. Spermiogenesis is a special stage of spermatogenesis, because of some technical reasons, most researchs for spermiogenesis are by gene-knockout technology. RNAi technology emerged in recent years was widely used because of its convenient and efficient, and it may provide us a new way of methods to study spermiogenesis. Here, we approach a new convenient and effective method of RNAi in vivo: RNAi in vivo by microinjection, then we utilized this method to desire whether lamin A/C played an important role in the mouse spermiogenesis.. We injected SiRNA of LMNA mixed with typan blue into seminiferous tubule of puberty mouse via intra-testicular injection, and later by observing the epididymal spermatozoa when the mouse was adult to study it's function during spermiogenesis.We found lots of deformity spermatozoa contained golbozoospermia because of the abnormal acrosome formation and nuclear shaping after lamin A/C was knocked-down. These results suggested that, LMNA may indeed involve in the formation of the acrosome, and it was effective and feasible to study functions of spermiogenesis-related genes by micro-injection combined with RNAi methods. Then we identified the physical interaction between lamin A/C and CLIP-170 in mouse testis by immunopreciptation coupled with MALDI-TOF. CLIP-170 has been identified associated with the sperm manchette, and CLIP-170 knockout mice were subfertile and produce sperm with abnormal heads. Acrosome-acroplaxome-machette cooperated with each other during the mouse sperm head shaping. Lamin A/C located in the acroplaxome, interacting with CLIP-170 in the manchette, made spermiogenesis carried out successfully[2].In summary, the construction of comparative proteome profile for mouse spermatogenesis showed us a completely new perspective to understand this mysterious process. In the later studies, we showed that the differentiated protein lamin A/C located in the acroplaxome in mouse sperm and spermatid cells and may involve in the spermiogenesis by interacting with CLIP-170.
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