H3K79 methylation directly precedes the histone-to-protamine transition in mammalian spermatids and is sensitive to bacterial infections.

In both mammalian and Drosophila spermatids, the completely histone-based chromatin structure is reorganized to a largely protamine-based structure. During this histone-to-protamine switch, transition proteins are expressed, for example TNP1 and TNP2 in mammals and Tpl94D in Drosophila. Recently, we demonstrated that in Drosophila spermatids, H3K79 methylation accompanies histone H4 hyperacetylation during chromatin reorganization. Preceding the histone-to-protamine transition, the H3K79 methyltransferase Grappa is expressed, and the predominant isoform bears a C-terminal extension. Here, we show that isoforms of the Grappa-equivalent protein in humans, rats and mice, that is DOT1L, have a C-terminal extension. In mice, the transcript of this isoform was enriched in the post-meiotic stages of spermatogenesis. In human and mice spermatids, di- and tri-methylated H3K79 temporally overlapped with hyperacetylated H4 and thus accompanied chromatin reorganization. In rat spermatids, trimethylated H3K79 directly preceded transition protein loading on chromatin. We analysed the impact of bacterial infections on spermatid chromatin using a uropathogenic Escherichia coli-elicited epididymo-orchitis rat model and showed that these infections caused aberrant spermatid chromatin. Bacterial infections led to premature emergence of trimethylated H3K79 and hyperacetylated H4. Trimethylated H3K79 and hyperacetylated H4 simultaneously occurred with transition protein TNP1, which was never observed in spermatids of mock-infected rats. Upon bacterial infection, only histone-based spermatid chromatin showed abnormalities, whereas protamine-compacted chromatin seemed to be unaffected. Our results indicated that H3K79 methylation is a histone modification conserved in Drosophila, mouse, rat and human spermatids and may be a prerequisite for proper chromatin reorganization.

Wolbachia induced cytogenetical effects as evidenced in Chorthippus parallelus (Orthoptera).

The cytoplasmic incompatibility induced by the bacterial endosymbiont Wolbachia is attributed to chromatin modification in the sperm of infected individuals and is only 'rescued' by infected females after fertilization. Chorthippus parallelus is a grasshopper with 2 subspecies that form a hybrid zone in the Pyrenees in which this Wolbachia-generated cytoplasmic incompatibility has recently been described. The analysis of certain cytogenetic traits (sex chromosome-linked heterochromatic bands, nucleolar organizing region expression, spermatid size and morphology, and number of chiasmata formed) in pure and hybrid Chorthippus parallelus that are infected and not infected by this bacterium indicates that the infection affects some of these traits and, in the case of the spermatids, reveals a synergism between the infection and the hybrid condition. These results are interpreted as being secondary effects of the chromatin modification induced by Wolbachia which thereby support this model of modification/rescue. The possible effects of these cytogenetic variations on affected individuals are also considered.

An ultrastructural study of vertical transmission of Rickettsia tsutsugamushi during oogenesis and spermatogenesis in Leptotrombidium pallidum.

Rickettsia tsutsugamushi in Leptotromibidium pallidum was observed by electron microscopy and rickettsiae were found in the various tissues and organs of both larvae and adults. Budding of rickettsiae, a manner of release from the host cells, was observed only in the rudiments of the reproductive organs in larvae. Oogonia and maturing oocytes in adult females and eggs after oviposition contained the microorganisms. In adult males, rickettsiae were also found in the spermatogonia, spermatocytes, and spermatids in the early stage of spermatogenesis, but were eliminated from these cells during maturation. Only the maturing spermatids, but not the eliminated rickettsiae, migrated to another rickettsia-free area of the testis, resulting in the separation of spermatids from rickettsiae and in the production of rickettsia-free spermatophores. Based on these observations, the mechanism of vertical transmission of the rickettsiae to the progeny occurs only in the female parents. Most rickettsiae in the somatic cells of larvae and adults were coccoid, but some rickettsiae in the ovary and the testis of adult mites showed bacillary forms and were enveloped by a membrane of unknown origin.

Sexual transmission of spotted fever group rickettsiae by infected male ticks: detection of rickettsiae in immature spermatozoa of Ixodes ricinus.

Electron microscopic evidence is provided showing that the newly described "Swiss agent," a spotted fever group rickettsia, is incorporated into the reproductive cells of its male tick vector, Ixodes ricinus. Rickettsiae were found in spermatogonia, spermatocytes, and maturing spermatids. The potential significance of these observations is briefly discussed in relation to published data on sexual transmission of rickettsiae by ticks.