Gene functional research using polyethylenimine-mediated in vivo gene transfection into mouse spermatogenic cells.
OBJECTIVE
To study polyethylenimine (PEI)-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method.


METHODS
PEI/DNA complexes were introduced into the seminiferous tubules of mouse testes using intratesticular injection. Transfection efficiency and speciality were analyzed on the third day of transfection with fluorescent microscopy and hematoxylin staining. The long-lasting expression of the GFP-NYD-SP12 fusion protein and its subcellular localization in spermatogenic cells at different stages were analyzed with fluorescent microscopy and propidium iodide staining.


RESULTS
With the mediation of PEI, the GFP-NYD-SP12 fusion gene was efficiently transferred and expressed in the germ cells (especially in primary spermatocytes). Transfection into Sertoli cells was not observed. The subcellular localization of the GFP-NYD-SP2 fusion protein showed dynamic shifts in spermatogenic cells at different stages during spermatogenesis.


CONCLUSIONS
PEI can efficiently mediate gene transfer into spermatocytes. Thus, it might be useful for the functional research of spermatogenic-cell specific genes such as the NYD-SP12 gene. In our study, the NYD-SP12 protein was visualized and was involved in the formation of acrosome during spermatogenesis. Our research will continue into the detailed function of NYD-SP12 in spermatocytes.
Lu
Li
L
Laboratory of Reproductive Medicine, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China.
Lin
Min
M
Xu
Min
M
Zhou
Zuo-Min
ZM
Sha
Jia-Hao
JH
D000818:Animals; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008297:Male; D051379:Mice; D011094:Polyethyleneimine; D013091:Spermatogenesis / Q000502:physiology; D014162:Transfection / Q000379:methods*; D033921:Vesicular Transport Proteins
D018398:Homeodomain Proteins; C474264:SPATA16 protein, human; D033921:Vesicular Transport Proteins; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine