Anti-BMP1 monoclonal antibody, clone 4F0

It has long been hypothesized that in fishes the contents of cortical granules are involved in the hardening of egg envelope following fertilization. We previously purified the egg envelope hardening initiation factor from the exudates released from activated medaka (Oryzias latipes) eggs and tentatively termed this protein alveolin. Alveolin is a member of the astacin metalloprotease family and was proposed to be a protease which hydrolyzes ZPB at one restricted position to allow starting cross-linking with ZPC. Here, we investigated the complete pathway from biosynthesis and accumulation to secretion of alveolin. A single alveolin transcript was detected only in ovarian preparations, confirming the specific expression of alveolin in the ovary. In situ hybridization indicated that the alveolin mRNA is already expressed in the very early previtellogenic oocytes. However, immunocytochemical studies revealed that the appearance of alveolin protein was delayed until the beginning of the vitellogenic stage. The cortical granules isolated from unfertilized eggs contained a high molecular weight form of glycosylated alveolin with a 50 kDa relative molecular mass. Hypotonic treatment burst isolated granules in vitro and transformed alveolin to a 21.5 kDa form, which is the same size as that of natural alveolin released from eggs upon fertilization. This transformation was inhibited in the presence of leupeptin and 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF), suggesting that a serine protease is involved in alveolin activation upon fertilization. Furthermore, the phylogenetic relationship of alveolin with other vertebrate astacin family members was analyzed. The result shows that alveolin and its teleostean homologs make a new group which is separate from either the hatching enzyme, meprin and BMP1/tolloid groups. (C) 2012 Elsevier Inc. All rights reserved.

In order to better understand the relationship between vegetation communities and water table in the uppermost portions (ephemeral-intermittent streams) of headwater systems, seasonal plot-based field characterizations of vegetation were used in conjunction with monthly water table measurements. Vegetation, soils, and water table data were examined to determine potential indicator species of near-surface water that could be used in rapid delineation of Streamside Management Zones (SMZs) by forest managers. Twelve watersheds were instrumented with three hundred screened wells, installed in grids of 25 per sub-watershed. Well locations were used to monitor water table and vegetation communities. Species were classified according to their wetland indicator status for Region 2; communities were evaluated using a prevalence index (PI). As part of a larger study, the uppermost reaches of the headwater systems were treated one of four treatments: (1) removal of all merchantable stems leaving understory intact with minimum surface soil disturbance (BMP1), (2) the same as treatment BMP1 with the addition of logging debris in the drainage channel (BMP2), (3) total harvest with no BMPs applied (clearcut) and (4) no harvest (reference). Post-harvest increases in water table ranged from 1.6 cm in BMP1 to 28.2 cm in clearcut treatments during 2008, from 10.5 cm in BMP1 to 54.2 cm in BMP2 during 2009. PI differed significantly between channel and hillslope positions and represent distinctive vegetation communities. Forest clearcutting affected vegetation communities through combined direct and indirect disturbances. PI in the clearcut did not respond directly to changes in water table. In the two treatments where BMPs were employed, changes in vegetative communities corresponded to both changes in water table and changes in the microclimate as a result of harvesting intensity and changing stand heterogeneity. A vegetative indicator analysis, based on the presence of saturated soil conditions and water table elevation, was used to determine potential indicators of the true hydrologic boundaries of the headwater streams. Three potential indicator species (Viola blanda, Ludwigia glandulosa, and Arundinaria gigantean Ssp. Tecta) were more prevalent within "wet" channel positions but exhibited less total frequency of occurrence across the study areas than the fern species Polystichum acrostichoides, which is locally used for rapid estimation of intermittent stream extent). The combined use of the strong indicator species identified in this study and the "fern line" used by local industry foresters provides a means for rapid assessment of hydrologically functional SMZs in these headwater streams. (C) 2012 Elsevier B.V. All rights reserved.