Expression profile of SPACA5/Spaca5 in spermatogenesis and transitional cell carcinoma of the bladder.

The majority of bladder cancer-associated mortalities are due to transitional cell carcinoma (TCC), which is the most prevalent and chemoresistant malignancy of the bladder. Sperm acrosome associated 5 (SPACA5)/Spaca5 is a sperm acrosome-associated, c-type lysozyme-like protein that has been recently identified, and has been designated as an attractive candidate antigen for cancer testis. In the present study, the expression profile of SPACA5/Spaca5 was analyzed in spermatogenesis and TCC of the bladder using diverse molecular and cellular biology methods. Using reverse transcription-polymerase chain reaction (RT-PCR) to analyze the multi-tissue distribution and temporal expression of SPACA5/Spaca5, the SPACA5/Spaca5 gene was determined to be generally not expressed in normal tissue, with the exception of the testis, and it could be detected at a low level on day 20 after birth in mouse testes and at a higher level on day 28. Immunohistochemistry staining revealed that the SPACA5/Spaca5 protein was exclusively observed in the elongated spermatid of the normal testes, and was ectopically expressed in the cytoplasm of TCC, while it was not expressed in normal bladder tissues. The frequency of SPACA5 messenger RNA was detected in 45% of TCC (9/20) by RT-quantitative PCR. Furthermore, SPACA5 protein was more frequently detected in high-grade than in low-grade tumors (61.54 vs. 30.00%, P=0.035). Accordingly, high SPACA5 staining scores were observed to be significantly associated with high-grade tumors (n=65, R=0.279, P=0.027). Collectively, our findings indicated that SPACA5/Spaca5 may be important in male spermatogenesis and may be used as a potential target for specific immunotherapy in patients suffering from TCC.

[Regulation of fertility by the small RNA pathway that defends the genome against transposons].

Small RNAs can silence transposons at transcriptional or post-transcriptional level. Newly identified small RNAs (e.g., piRNAs and endo-siRNAs) can repress the activity of transposons. Drosophila piRNAs, mouse piRNAs, mouse endo-siRNAs, and their related proteins are expressed primarily within the germline. This suggests that the small RNA pathway plays an important role in transposon silencing of the germline. Recent studies revealed the connection between small RNAs, transposon silencing, and the regulation of fertility, but the mechanism has not been clarified in detail. In this review, the advances in the control of genomic transposon activity and the regulation of fertility by the small RNA pathway were summarized and discussed.

Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia.

AIM: To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. METHODS: Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction (RT-PCR), quantitative RT-PCR (QRT-PCR), immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. RESULTS: VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. CONCLUSION: The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.

[DNA microarray and its application in andrological research].

This article briefly retrospect the development of microarray, introduces the basic working procedures and the current challenges of DNA microarray, and reviews its application to andrological research, as on the testis, spermatogenic cells, epididymis and sperm. We hope it could play a directive role in the studies of male infertility.