In most amoeboid cells, the main protein involved in motility is actin. Nematode sperm are an exception, and their amoeboid motility is based on major sperm protein (MSP). We have studied the localization of actin and MSP in spermatids and spermatozoa of Graphidium strigosum (Dujardin, 1845), a species which has elongate male germ cells in which organelles are easily identified. Electrophoreses of G. slrigosum sperm proteins indicate that the main protein band, about 15 kDa in molecular weight, is specifically recognized by an anti-MSP polyclonal antibody developed against MSP of Caenorhabditis elegans (Burke and Ward 1983). Actin is present in small quantities. Immunocytochemical observations reveal that actin and MSP have an identical localization in precise areas of the male germ cells. Spermatids are labelled as dots around a central unlabelled zone, and spermatozoa are labelled only at the level of the anterior cap. Observations in G. strigosum are similar to that previously obtained in Heligmosomoides polygyrus (Mansir and Justine 1996). Co-localization of actin and MSP in the anterior cap of the spermatozoon, the region associated with pseudopod production, does not demonstrate directly that actin is involved in amoeboid movements, but shows that the role of actin in the cytoskeleton of nematode sperm should be re-investigated.
The aim of our study was to test the in fluence of short exposure (6 h) of preimplantation rabbit embryos to elevated temperatures (41.5 ºC or 42.5 ºC) in vitro on their developmental capacity. Fertilized eggs recovered from female oviducts at the pronuclear stage (19 hpc) were cultured at standard temperature (37.5 ºC) until the morula stage (72 hpc). Afterwards, the embryos were divided into two groups, cultured for 6 h either at hyperthermic (41.5 ºC or 42.5 ºC) or standard temperature (control 37.5 ºC), post-incubated overnight (16-20 h) at 37.5 ºC and then evaluated for developmental stages, apoptosis (TUNEL), proliferation (cell number), actin cytoskeleton and presence of heat-shock proteins Hsp70. It was observed that hyperthermia at 41.5 ºC did not alter progression of embryos to higher preimplantation stages (expanded and hatching/hatched blastocysts), rate of apoptosis, total cell number of blastocysts and structure of actin filament compared to 37.5 ºC. We stern-blotting revealed the presence of heat stress-induced 72 kDa fraction of Hsp70 proteins in granulosa cells (exposed to 41 ºC) and embryos (exposed to 41.5 ºC). Following the elevation of temperature to 42.5 ºC embryo development was dramati cally compromised. The embryos were arrested at the morula or early blastocyst stage, showed an increased rate of apoptosis and decreased total cell number compared to control. The structure of actin filaments in most of blastomeres was damaged and such blastomeres often contained apoptotic nuclei. In this group a presence of heat-stress-induced fraction of Hsp70 proteins had not been confirmed. This is the first report demonstrating a threshold of thermotolerance of rabbit preimplantation embryos to hyperthermic exposure in vitro. A detrimental effect of higher temperature on the embryo is probably associated with the loss of their ability to produce Hsp70 de novo, which leads to cytoskeleton alterations and enhanced apoptosis., A. V. Makarevich, L. Olexiková, P. Chrenek, E. Kubovičová, K. Fréharová, J. Pivko., and Obsahuje bibliografii a bibliografické odkazy