Open AccessGene functional research using polyethylenimine‐mediated in vivo gene transfection into mouse spermatogenic cells
Author(s) -
Lu Li,
Lin Min,
Xu Min,
Zhou ZuoMin,
Sha JiaHao
Publication year - 2006
Publication title -
asian journal of andrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.701
H-Index - 74
eISSN - 1745-7262
pISSN - 1008-682X
DOI - 10.1111/j.1745-7262.2006.00089.x
Subject(s) - transfection , microbiology and biotechnology , propidium iodide , sertoli cell , green fluorescent protein , biology , spermatogenesis , fusion protein , reporter gene , gene , gene expression , genetics , recombinant dna , apoptosis , programmed cell death , endocrinology
Aim: 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 subcelluar 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. Conclusion: 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.