Volume 11, Issue 3 (May 2009) 11, 342–350; 10.1038/aja.2008.54
Interstitial tissue-specific gene expression in mouse testis by intra-tunica albuguineal injection of recombinant baculovirus
Hyun Jung Park1,*, Won Young Lee1,2,*, Jin Hoi Kim3, Jae Hwan Kim3, Hun Jong Jung4, Nam Hyung Kim5, Bo Kyung Kim6 and Hyuk Song1
1 Department of Animal Science, College of Natural Science, Konkuk University, Chung-ju 380-701, Korea
2 CHA Stem Cell Institute, Graduate School of Life Science and Biotechnology, Pochon CHA University, Seoul 135-907, Korea
3 Department of Animal Biotechnology, College of Animal Biotechnology, Konkuk University, Seoul 143-701, Korea
4 Occupation and Environment Department, Medical School, Konkuk University, Chung-ju 380-704, Korea
5 Department of Animal Science, College of Agriculture, Chungbuk National University, Choung-ju, Chung-buk 361-763, Korea
6 Department of Physiology, Medical School, Konkuk University, Chung-ju 380-701, Korea
* These two authors contributed equally to this work.
Correspondence: Prof. Hyuk Song, Department of Animal Science, College of Natural Science, Konkuk University, Chung-ju 380-701, Korea. Fax: +82-43-842-3522 E-mail: email@example.com
Received 1 July 2008; Revised 19 September 2008; Accepted 24 October 2008; Published online 26 January 2009
The purpose of this study is to establish a gene delivery system for interstitial tissue-specific protein expression in mice testes using modified recombinant baculovirus. Green fluorescent protein (GFP)-expressing recombinant baculovirus (GFP-baculovirus), in which the insect cell-specific polyhedron promoter was replaced by the cytomegalovirus (CMV)-IE promoter, was used to transfect testicular cells in vitro, and for intra-tunica albuguineal injection of the interstitial tissue of the testis. GFP expression was monitored in frozen testes sections by fluorescence microscopy. Expression of GFP in testicular tissues was also assessed by reverse transcription polymerase chain reaction (RT-PCR), and protein expression was assessed by Western blot. Testicular cells in vitro were infected efficiently by modified recombinant GFP-baculovirus. Intra-tunica albuguineal injection of GFP-baculovirus into the mouse testis resulted in a high level of GFP expression in the interstitial tissues. RT-PCR analysis clearly showed GFP gene expression in the testis, particularly interstitial tissues. Intra-tunica albuguineal injection of a modified baculovirus that encoded recombinant rat insulin-like growth factor binding protein (IGFBP)-5 resulted in an increase in IGFBP-5 in testis and semen. In conclusion, we have developed an efficient delivery system for gene expression in vivo in testicular cells, particularly cells of the interstitial tissue using intra-tunica albuguineal injection of a modified recombinant baculovirus. This method will be particularly relevant for application that requires gene delivery and protein expression in the testicular cells of the outer seminiferous tubule of the testis.
Keywords: intra-tunica albuguineal injection, recombinant baculovirus, testis gene delivery