Plasmin decreases the BH3-only protein BimEL via the ERK1/2 signaling pathway in hepatocytes

Since the signal transduction mechanisms responsible for liver regeneration mediated by the plasminogen/plasmin system remain largely undetermined, we have investigated whether plasmin regulates the pro-apoptotic protein Bim(EL) in primary hepatocytes. Plasmin bound to hepatocytes in part via its lysine binding sites (LBS). Plasmin also triggered phosphorylation of ERK1/2 without cell detachment. The plasmin-induced phosphorylation of ERK1/2 was inhibited by the LBS inhibitor epsilon-aminocaproic acid (EACA), the serine protease inhibitor aprotinin, and the MEK inhibitor PD98059. DFP-inactivated plasmin failed to phosphorylate ERK1/2. Plasmin temporally decreased the starvation-induced expression of Bim(EL) and activation of caspase-3 via the ERK1/2 signaling pathway, resulting in an enhancement of cell survival. The amount of mRNA for Bim increased 1 day after the injection of CCl(4) in livers of plasminogen knockout (Plg-KO) and the wild-type (WT) mice. The increase in Bim(EL) protein persisted for at least 7 days post-injection in livers of Plg-KO mice, whereas WT mice showed an increase in Bim(EL) protein 1 day after the injection. Plg-KO and WT mice showed notable phosphorylation of ERK1/2 7 and 3 days after the injection of CCl(4), respectively. Our data suggest that the plasminogen/plasmin system could decrease Bim(EL) expression via the ERK1/2 signaling pathway during liver regeneration.     

Influence of Lactic Acid Bacteria on Longevity of Caenorhabditis elegans and Host Defense against Salmonella enterica Serovar Enteritidis

This study aimed to develop a convenient model to investigate the senescence of host defenses and the influence of food and nutrition. A small soil nematode, Caenorhabditis elegans, was grown for 3 days from hatching on a lawn of Escherichia coli OP50 as the normal food source, and subsequently some of the nematodes were fed lactic acid bacteria (LAB). The life spans of worms fed LAB were significantly longer than the life spans of those fed OP50. To investigate the effect of age on host defenses, 3- to 7-day-old worms fed OP50 were transferred onto a lawn of Salmonella enterica serovar Enteritidis for infection. The nematodes died over the course of several days, and the accumulation of salmonella in the intestinal lumen suggested that the worms were infected. The 7-day-old worms showed a higher death rate during the 5 days after infection than nematodes infected at the age of 3 days; no clear difference was observed when the worms were exposed to OP50. We then investigated whether the LAB could exert probiotic effects on the worms' host defenses and improve life span. Seven-day-old nematodes fed LAB from the age of 3 days were more resistant to salmonella than worms fed OP50 until they were infected with salmonella. This study clearly showed that LAB can enhance the host defense of C. elegans and prolong life span. The nematode appears to be an appropriate model for screening useful probiotic strains or dietetic antiaging substances.     

Changes in the components of biliary and plasma lipids in selenium-deficient rats

We constructed a chronic oxidative stress model in which Se-deficient diet was fed to male Wister rats for 8 weeks. As expected, effects of oxidative damage, including Fe accumulation and increase in peroxidized lipids, were identified in the liver owing to the lack of glutathione peroxidase. Although the oxidative stress caused Fe accumulation in the liver, the Fe concentration in bile of the SeD rat was almost the same as that in the control rats. The constant excretion of Fe into bile supported the Fe accumulation in the liver. No differences were observed in the principal components of biliary lipids, i.e., bile acids, phospholipids, and cholesterol, between the two groups; moreover, these trends were also reflected in the plasma. Due to the trapping of reactive oxygen species, only bilirubin concentrations in the bile and plasma were decreased in the SeD group, when compared with those in the control group. Measurement of bilirubin concentration may be used as a supplemental oxidative stress marker.     

Proliferation of germ cells during gonadal sex differentiation in medaka: Insights from germ cell-depleted mutant zenzai

The proliferation of germ cells becomes sexually dimorphic during gonadal sex differentiation, although the underlying dynamics of this are not well understood in vertebrates. By tracing GFP-labeled germ cells in vivo and analyzing the germ cell-depleted mutant, zenzai, we show that the proliferation and differentiation of germ cells are regulated in a sexually dimorphic manner in the teleost fish medaka. In the undifferentiated gonads, germ cells resume proliferation by slow intermittent division (type I), producing isolated daughter cells. While germ cells in the male gonads continue this mode of proliferation, some germ cell fractions in the female gonads initiate two to four rounds of continuous division (type II), forming cysts of four, eight, or sixteen cells, which subsequently enter meiosis synchronously. Thus, female germ cells become differentiated much earlier than do male germ cells. In the zenzai mutant, a defect in slow intermittent division eventually leads to the depletion of germ cells in the adult gonads in both sexes, despite the fact that cyst-forming division is unaffected. This argues that slow intermittent division is essential for the maintenance of germ cells. The proliferation and differentiation of germ cells are thus important components of gonadal sex differentiation in vertebrates.     

Mutations affecting gonadal development in Medaka, Oryzias latipes

A gonad is formed from germ cells and somatic mesodermal cells through their interactions. Its development is coupled with the determination and differentiation of the sex and sex-associated traits. We carried out a large-scale screening of Medaka mutants in which gonadal development is affected. Screening was performed on larvae at 8 days posthatching for abnormal abundance and/or distribution of germ cells detected by the in situ hybridization for olvas (Medaka vasa). We describe here 16 mutants of 13 genes, which are classified into four groups. Group 1, consisting of four mutants of three genes kon, tot) characterised by an increase in germ cell number. An adult tot homozygote fish has the characteristic feature of possessing hypertrophic gonads filled with immature oocytes. Group 2, represented by a single gene (zen) mutant characterized by a gradual loss of germ cells. Group 3, consisting of four mutants of distinct genes (eko, eki, sht, ano) showing irregular clustering of germ cells. Group 4, consisting of seven mutants of five genes (arr, hyo, mzr, hdr, fbk) showing fragmented clusters of germ cells. In some mutants belonging to Groups 1, 3 and 4, the expression level of ftz-f1 (sf-1/Ad4BP) in gonadal somatic cells significantly decreased, suggesting that interaction between somatic and germ cells is affected.