Small-scale temporal variation in propagule supply of an intertidal red alga

Information on the variability in supply of algal propagules is scarce, hindered by the difficulty in identifying propagules, but this variability may affect the distribution and abundance of algal assemblages. This study examined the small-scale (½ hourly to hourly) temporal variation in propagule supply of Chondrus verrucosus (Gigartinaceae, Rhodophyta) over a dense, isolated bed in south-eastern Japan in summer and winter of 1999. Either 0.5 litre scoop samples or 5 litre pump samples were collected ½ hourly to hourly over 13, 22.5, and 30 h on three occasions in summer (June & July) and 32 h on one occasion in winter (December). Sampling was conducted around either the new moon (two occasions in summer) or full moon (one occasion in both summer and winter) and incorporated full tidal sequences including daytime (summer) and nighttime (winter) low-low (LL) tides. Chondrus verrucosus was the only red alga with spores within the size range of 15–20 μm that was fertile in the study area and surrounds at the time of sampling facilitating identification of spores. Spores in scoop samples were settled onto Petri dishes and identified on the basis of cell shape, colour and size. Pump samples were filtered onto transparent membrane filters and identified using epifluorescence microscopy: C. verrucosus spores fluoresced bright yellow and were easily distinguished from other micro-organisms of similar size, which fluoresced red or green. Results showed that while propagules could be found in the water column at most times, propagule supply of C. verrucosus was greatest during the 1–2 h period following LL tides. Variability in propagule supply was less than in previous studies examining surface or offshore waters. Spore release is thought to be stimulated by either desiccation or salinity changes associated with periods of emersion at low tide followed by re-immersion on incoming tides.

Turnover of bone marrow-derived cells in the irradiated mouse cornea

In light of an increasing awareness of the presence of bone marrow (BM)-derived macrophages in the normal cornea and their uncertain role in corneal diseases, it is important that the turnover rate of these resident immune cells be established. The baseline density and distribution of macrophages in the corneal stroma was investigated in Cx3cr1gfp transgenic mice in which all monocyte-derived cells express enhanced green fluorescent protein (eGFP). To quantify turnover, BM-derived cells from transgenic eGFP mice were transplanted into whole-body irradiated wild-type recipients. Additionally, wild-type BM-derived cells were injected into irradiated Cx3cr1+/gfp recipients, creating reverse chimeras. At 2, 4 and 8 weeks post-reconstitution, the number of eGFP+ cells in each corneal whole mount was calculated using epifluorescence microscopy, immunofluorescence staining and confocal microscopy. The total density of myeloid-derived cells in the normal Cx3cr1+/gfp cornea was 366 cells/mm2. In BM chimeras 2 weeks post-reconstitution, 24% of the myeloid-derived cells had been replenished and were predominantly located in the anterior stroma. By 8 weeks post-reconstitution 75% of the myeloid-derived cells had been replaced and these cells were distributed uniformly throughout the stroma. All donor eGFP+ cells expressed low to moderate levels of CD45 and CD11b, with approximately 25% coexpressing major histocompatibility complex class II, a phenotype characteristic of previous descriptions of corneal stromal macrophages. In conclusion, 75% of the myeloid-derived cells in the mouse corneal stroma are replenished after 8 weeks. These data provide a strong basis for functional investigations of the role of resident stromal macrophages versus non-haematopoietic cells using BM chimeric mice in models of corneal inflammation.