Expression of an avian protamine in transgenic mice disrupts chromatin structure in spermatozoa.

The objective of this study was to determine the consequences of disrupting spermatozoal chromatin condensation on spermatozoal development and function. The avian protamine, galline, was targeted to spermatids of transgenic mice using the mouse protamine 1 gene promoter. Three transgenic mouse lines were established that expressed galline mRNA at 65%, 120%, and 185% of the level found in rooster testis. Galline mRNA accumulated in round spermatids to levels similar to that of mouse protamine and, as with the mammalian counterpart, translation was delayed until the elongating spermatid stage. Protein gels revealed that galline accumulated in mature spermatozoa whereas mouse protamines were reduced, suggesting that galline competes with protamines for binding to spermatozoal DNA. Acridine orange binding analysis indicated that DNA of the transgenic spermatozoa was not as tightly packed as that of controls. This was corroborated by electron microscopy, which revealed disruption of the normal dense chromatin structure of spermatozoal heads. Despite these perturbations of chromatin condensation, the transgenic spermatozoa were functionally normal, as the majority of transgenic mice had normal fertility. However, in mice that expressed excessive galline, there was a gradual destruction of seminiferous tubules leading to infertility. Our findings suggest that very precise packaging of DNA in germ cells may not be essential for subsequent unpackaging in the pronucleus of fertilized eggs and for subsequent normal development of the embryo.