[Frontiers in Bioscience E2, 187-194, January 1, 2010]

LM23 is essential for spermatogenesis in Rattus norvegicus

Mei-ling Liu1,2, Yi-ming Cheng1, Meng-chun Jia2

1Graduate School of Peking Union Medical College, #5 Dong Dan San Tiao, Beijing, 100005, China,2 Department of Reproductive Endocrinology, National Research Institute for Family Planning, No. 12 Da Hui Si, Hai Dian District, Beijing 100081, P.R. China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Experimental animals
3.2. Construction of fusion protein expression vector
3.3. Lentivirus construction
3.4. Generation of high-titer lentivirus
3.5. RNAi in vivo
3.6. Histological and TUNEL analysis
3.7. Real-time PCR
3.8. Statistical analysis
4. Results
4.1. Lentivirus-mediated RNAi efficiently inhibited testicular LM23 expression in vivo
4.2. Impaired spermatogenesis in the LM23 knockdown Rattus norvegicus
4.3. Increased apoptosis in LM23-knockdown Rattus norvegicus
4.4. LM23-regulated genes in testes
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

LM23 is a gene with testis-specific expression in Rattus norvegicus. To reveal the function of LM23 in the testis, we used lentivirus-mediated RNA interference (RNAi) to knock down LM23 expression in a tissue-specific manner in vivo. A lentiviral vector expressing a short hairpin RNA (shRNA) targeting LM23 was microinjected into the efferent ducts of R. norvegicus testes. The expression of LM23 in the treated testes was significantly knocked down compared with controls. These LM23-shRNA testes contained germ cells arrested at the spermatocyte stage, and showed increased apoptosis and disregulation of some meiotic genes. The results demonstrate the validity of the RNAi approach for targeting LM23 and reveal that LM23 expression in the testis is crucial for meiosis during spermatogenesis in R. norvegicus.