In vivo gene transfer into the testis by electroporation and viral infection--a novel way to study testis and sperm function.
The study of gene function in testis and sperm has been greatly assisted by creation of transgenic mice by injection of a transgene into the fertilised egg. However this approach is costly and laborious and is not applicable to other species of importance for the study of sperm function, such as the hamster. We have investigated alternative ways of expressing transgenes in mouse and hamster testis and sperm by in vivo gene transfer. DNA expression constructs were introduced into the testis by injection of DNA followed by electroporation, or by injection of a lentiviral vector. Expression of fluorescent proteins was assessed by fluorescence microscopy. In vivo gene transfer by electroporation led to expression of a fluorescent reporter protein and a fluorescently tagged version of sperm protein phospholipase C zeta in hamster and mouse testis and epididymal sperm. In vivo gene transfer by lentiviral infection led to high level expression of a fluorescent reporter protein in male germ cells. In conclusion, in vivo gene transfer offers a novel way to study gene function in testis and sperm and may also have potential as a way of creating transgenic versions of important model organisms such as the hamster.
Parrington
J
J
Department of Pharmacology, University of Oxford, Oxford, UK. john.parrington@pharmacology.oxford.ac.uk
Coward
K
K
Hibbitt
O
O
Kubota
H
H
Young
C
C
McIlhinney
J
J
Jones
O
O
D000818:Animals; D006224:Cricetinae; D004247:DNA / Q000008:administration & dosage*; D018274:Electroporation; D005456:Fluorescent Dyes; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008815:Mice, Inbred Strains; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D013094:Spermatozoa / Q000502:physiology* / Q000821:virology; D013737:Testis / Q000502:physiology* / Q000821:virology; D014161:Transduction, Genetic / Q000379:methods*
D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D004247:DNA