We assessed the levels and localization of the actin filament-severing protein scinderin, in fetal and adult bovine testes, and in spermatozoa during and following the epididymal transit. We performed immunoblots on seminiferous tubules and interstitial cells isolated by enzymatic digestion, and on bovine chromaffin cells, spermatozoa, aorta, and vena cava. Immunoperoxidase labeling was done on Bouin's perfusion-fixed testes and epididymis tissue sections, and on spermatozoa. In addition, immunofluorescence labeling was done on spermatozoa. Immunoblots showed one 80-kDa band in chromaffin cells, fetal and adult tubules, interstitial cells, spermatozoa, aorta, and vena cava. Scinderin levels were higher in fetal than in adult seminiferous tubules but showed no difference between fetal and adult interstitial cells. Scinderin levels were higher in epididymal than in ejaculated spermatozoa. Scinderin was detected in a region corresponding with the subacrosomal space in the round spermatids and with the acrosome in the elongated spermatids. In epididymal spermatozoa, scinderin was localized to the anterior acrosome and the equatorial segment, but in ejaculated spermatozoa, the protein appeared in the acrosome and the post-equatorial segment of the head. In Sertoli cells, scinderin was detected near the cell surface and within the cytoplasm, where it accumulated near the base in a stage-specific manner. In the epididymis, scinderin was localized next to the surface of the cells; in the tail, it collected near the base of the principal cells. In Sertoli cells and epididymal cells, scinderin may contribute to the regulation of tight junctional permeability and to the release of the elongated spermatids by controlling the state of perijunctional actin. In germ cells, scinderin may assist in the shaping of the developing acrosome and influence the fertility of the spermatozoa.
3At the time of the Great East Japan earthquake and tsunami (March 2011), the authors developed a web-based information and communications technology (ICT)-based blood pressure (BP) monitoring system (the Disaster CArdiovascular Prevention [DCAP] Network) and introduced it in an area that was catastrophically damaged (Minamisanriku town) to help control the survivors' BP. Using this system, home BP (HBP) was monitored and the data were automatically transmitted to a central computer database and to the survivors' attending physicians. The study participants, 341 hypertensive patients, continued to use this system for 4 years after the disaster and all of the obtained HBP readings were analyzed. This DCAP HBP-guided approach helped achieve a decrease in the participants' HBPs (initial average: 151.3AE20.0/86.9AE10.2 mm Hg to 120.2AE12.1/ 70.8AE10.2 mm Hg) over the 4 years. In addition, the amplitude of seasonal BP variation was suppressed and the duration from the summer lowest HBP values to the winter peak HBP values was gradually prolonged. This ICT-based approach was useful to achieve strict HBP control and minimize the seasonal BP variation even in a catastrophically damaged area during a 4-year period after the disaster, suggesting that this approach could be a routine way to monitor BP in the community.
To elucidate the significance of alpha- and alpha+ isoforms of the tight-junction-associated protein ZO-1 in Sertoli cell tight junction regulation, taking into consideration that different isoforms are expressed in cells with different junctional morphologies, we investigated whether alpha- and alpha+ are differentially associated with junctions forming the continuous occluding zonules responsible for the blood-testis barrier, and/or with junctions forming the focal discontinuous occluding zonules. In addition, since Sertoli cells contact Sertoli cells and germ cells, we investigated whether each isoform is differentially associated with distinct classes of germ cells. Our immunoblot analyses of isolated seminiferous tubules, using affinity-purified polyclonal antibodies recognizing rat and human alpha- and alpha+, showed that guinea pig testis contained the two ZO-1 isoforms initially described in rat and human kidneys, and that alpha+ and alpha- were predominantly expressed during puberty and adulthood, respectively, indicating that alpha+ was predominant during periods of increased junction assembly/disassembly. We used the same antibodies and immunoperoxidase labeling on fetal, neonatal, pubertal, and adult guinea pig testes sections. Both isoforms were expressed at the site of Sertoli cell-Sertoli cell and Sertoli cell-germ cell junctions in the seminiferous epithelium, before and after birth, and both were localized in continuous and in discontinuous tight junctions. However, the distribution of alpha- and alpha+ was not the same in different locations of the tight junctions. Only alpha- was incorporated into junctions joining the Sertoli cells to all classes of germ cells. The alpha+ involved junctions joining Sertoli cells to particular classes of germ cells, suggesting that Sertoli cell expression of ZO-1 isoforms could be regulated by unique germ cell-Sertoli cell contacts. Conversely, we found a correspondence between the distribution of F-actin and ZO-1alpha+, indicating that the spatial organization of the subsurface actin accompanying cell junctions may affect alpha+/alpha(-)-plasma membrane association.
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