Long-Term Field Experiment for Monitoring Soil Carbon Content in Japanese Grasslands: Initial Data from 2010 to 2012

A long-term field experiment for monitoring soil carbon content in Japanese grasslands started in 2010 to investigate the changes in soil carbon content and the effect of composted livestock manure application. We established grassland plots with 3 levels of manure application treatment at 10 sites. Bulk density values in many sites had wide inter-replicate and inter-annual variability. It is reasonable to suppose that the variability in the bulk density reflect spatial variability of physical properties within the grasslands because the annual trends of the bulk density values were not consistent. Organic carbon concentration tended to increase yearly in the surface layer (0–5 cm), whereas those for the subsoil layer (5–30 cm) stayed relatively constant. The organic carbon concentration in the surface layer tended to increase with increasing latitude and the amount of manure applied. When data from all the sites were taken into account, carbon content also tended to increase over time following grassland renovation. These results indicate that Japanese grasslands have the potential to sequester organic carbon. The monitoring has just begun, and it is important to continue the effort to achieve the goals of this study

Appearance of Dark Keratinocytes Following Intracutaneous Injection of Cholera Toxin in Mouse Skin

Intracutaneous injection of cholera toxin (CT), exotoxin of Vibrio cholerae, induces epidermal hyperplasia in mice, rats, and hamsters. In the work reported here we found that, like other hyperplasiogenic compounds such as 12-O-tetradecanoylphorbol-13-acetate which are tumor promoters, CT induces dark basal keratinocytes (dark cells) in the epidermis of mice. These are distinct from other epidermal cells since they contain dense cytoplasm rich in ribosomes and tonofilaments. This was demonstrated by electron microscopy and by toluidine blue staining of paraffin- or Epon-embedded sections. They comprised 3.1% of interfollicular basal cells 24–64 h after injection of 1 ng CT as compared with 0.5% in saline-injected skin. It was found by autoradiography of paraffin sections that about 47.2% of dark cells were labeled with [3H]thymidine at these times, while under the same conditions, labeling indices of basal cells were about 30% at the peaks. These results are discussed in relation to tumor promotion in two-stage carcinogenesis of mouse skin

Molecular cloning and characterization of a novel splicing variant of the Kir3.2 subunit predominantly expressed in mouse testis

One of the features of weaver mutant mice is male infertility, which suggests that Kir3.2, a G-protein-gated inwardly rectifying K+ channel subunit, may be involved in spermatogenesis. Therefore, we have characterized the Kir3.2 isoform in mouse testis using immunological, molecular biological and electrophysiological techniques.Testicular membrane contained a protein that was recognized by the antibody specific to the C-terminus of Kir3.2c (aG2C-3). Its molecular mass was ≈45 kDa, which was smaller than that of Kir3.2c (≈48 kDa). The immunoprecipitant obtained from testis with aG2C-3 contained a single band of the 45 kDa protein, which could not be detected by the antibody to the N-terminus common to the known Kir3.2 isoforms (aG2N-2).A novel alternative splicing variant of Kir3.2, designated Kir3.2d, was isolated from a mouse testis cDNA library. The cDNA had an open reading frame encoding 407 amino acids, whose molecular mass was calculated to be ≈45 kDa. Kir3.2d was 18 amino acids shorter than Kir3.2c at its N-terminal end, which was the only difference between the two clones. The 18 amino acid region possesses the epitope for aG2N-2.In heterologous expression systems of both Xenopus oocytes and mammalian cells (HEK 293T), Kir3.2d either alone or with Kir3.1 exhibited G-protein-gated inwardly rectifying K+ channel activity.Prominent Kir3.2d immunoreactivity in the testis was detected exclusively in the acrosomal vesicles of spermatids, while Kir3.1 immunoreactivity was diffuse in the spermatogonia and spermatocytes. These results indicate the possibility that the testicular variant of Kir3.2, Kir3.2d, may assemble to form a homomultimeric G-protein-gated K+ channel and be involved in the development of the acrosome during spermiogenesis