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This dissertation, Spatiotemporal expression of VAD1.2/AEP2 in spermatogenesis by Yuen-tsung, Tam, 譚婉頌, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled SPATIOTEMPORAL EXPRESSION OF VAD1.2/AEP2 IN SPERMATOGENESIS Submitted by TAM YUEN TSUNG for the Degree of Master of Philosophy at the University of Hong Kong in August 2007 Spermatogenesis is a process in which undifferentiated male germ cells undergo mitotic and meiotic divisions, followed by a dramatic morphological reorganization which is known as spermiogenesis to generate mature spermatozoa that are capable of fertilizing an oocyte. Maturation and differentiation of male germ cells take place within the seminiferous tubules of testis throughout the lifespan of adult males. To have a better understanding on spermatogenesis, we used vitamin A-deficiency (VAD) rat model to synchronize spermatogenesis in rat testis and mRNA differential display to profile the gene expression patterns in retinol re-initiated (PVA-VAD) spermatogenesis. Twelve differentially expressed cDNA fragments including VAD1.2 (AEP2, acrosome-expressing protein 2) was isolated. Bioinformatics analysis suggested that mouse VAD1.2/AEP2 protein shared 74% and 38% identity with rat and human counterparts. Northern blotting, semi-quantitative RT-PCR, real-time PCR and in situ hybridization studies suggested that VAD1.2/AEP2 gene was strongly expressed in the adult testis at the spermatid stage. VAD1.2/AEP2 protein was also detected in human, monkey and pig spermatids, and in rodent testis from day 25 postnatally. Furthermore, transmission electron microscopy (TEM) on rodent testes suggested that VAD1.2/AEP2 protein was expressed in the acrosomal cap of developing spermatids. Co-immunoprecipitation experiments identified β-actin and syntaxin 1 as potential interacting proteins that interact with VAD1.2/AEP2 in mouse, rat and human testis lysates. Our results demonstrated that VAD1.2/AEP2 was evolutionary conserved acrosome-expressing molecules that interact with β-actin and syntaxin 1 for cellular functions. Therefore, it is suggested that VAD1.2/AEP2 may play an important role(s) on vesicle protein binding during acrosome formation in spermatids. (251 words) DOI: 10.5353/th_b3955807 Subjects: Gene expressionSpermatogenesis - GeneticsSpermatogenesis in animalsRats

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