Distinct sequential cell behaviours direct primitive endoderm formation in the mouse blastocyst

The first two lineages to differentiate from a pluripotent cell population during mammalian development are the extraembryonic trophectoderm (TE) and the primitive endoderm (PrE). Whereas the mechanisms of TE specification have been extensively studied, segregation of PrE and the pluripotent epiblast (EPI) has received comparatively little attention. A current model of PrE specification suggests PrE precursors exhibit an apparently random distribution within the inner cell mass of the early blastocyst and then segregate to their final position lining the cavity by the late blastocyst. We have identified platelet-derived growth factor receptor alpha (Pdgfralpha) as an early-expressed protein that is also a marker of the later PrE lineage. By combining live imaging of embryos expressing a histone H2B-GFP fusion protein reporter under the control of Pdgfra regulatory elements with the analysis of lineage-specific markers, we investigated the events leading to PrE and EPI lineage segregation in the mouse, and correlated our findings using an embryo staging system based on total cell number. Before blastocyst formation, lineage-specific factors are expressed in an overlapping manner. Subsequently, a gradual progression towards a mutually exclusive expression of PrE- and EPI-specific markers occurs. Finally, cell sorting is achieved by a variety of cell behaviours and by selective apoptosis.     

The color of mice: in the light of GFP-variant reporters

The mouse currently represents the premier model organism for mammalian genetic studies. Over the past decade the production of targeted and transgenic lines of mice has become commonplace, with current technology allowing the creation of mutations at base pair resolution. Such genome modifications are becoming increasingly elaborate and often incorporate gene-based reporters for tagging different cellular populations. Until recently, lacZ, the bacterial beta-galactosidase gene has been the marker of choice for most studies in the mouse. However, over the past 3 years another valuable reporter has emerged, and its attractiveness is reflected by an explosion in its use in mice. Green fluorescent protein (GFP), a novel autofluorescent genetic reporter derived from the bioluminescent jellyfish Aequorea victoria, currently represents a unique alternative to other gene-based reporters in that its visualization is non-invasive and so can be monitored in real-time in vitro or in vivo. It has the added advantage that it can be quantified by, for example, flow cytometry, confocal microscopy, and fluorometric assays. Several mutants of the original wild-type GFP gene that improve thermostability and fluorescence have been engineered. Enhanced GFP is one such variant, which has gained popularity for use in transgenic or targeted mice. Moreover, various GFP spectral variants have also been developed, and two of these novel color variants, enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP), can also be used in mice. Since the spectral profiles of the ECFP and EYFP color variants are distinct and non-overlapping, these two reporters can be co-visualized, and are therefore ideal for in vivo double-labeling or fluorescent energy transfer analyses. The use of GFP and its color variants as reporters provides an unprecedented level of sophistication and represents the next step in mouse genome engineering technology by opening up the possibility of combinatorial non-invasive reporter usage within a single animal.     

Shedding light on bioscience. Symposium on Optical Imaging: Applications to Biology and Medicine

This dynamic symposium, held on 11-16 February 2003 in Taos, New Mexico, was the first Keystone meeting to focus on optical techniques and their use in biology and medicine. It was organized by D. Becker, D. Farkas and S. Fraser and attracted almost 100 participants from both academia and industry. Fluorescence imaging and its applications, ranging from nano-bioscience to small-animal imaging and imaging of disease progression in humans, were the main topics, with opportunities for further discussion in the cantinas of the town and on the ski slopes of Taos mountain.     

Pattern of Resource Allocation of Six Plantago Species with Different Breeding Systems

Plantago exhibit great deal of differences in the breeding system. The reproductive effort calculated on the basis of, (i) dry biomass of foliar and floral parts and (ii) seed output-weight (mg) per unit leaf area (cm²), exhibits relation with breeding system. The predominantly inbreeding taxa invest higher reproductive effort compared to their outbreeding allies. In terms of sex allocation strategies, the outbreeding species like P. lanceolata, P. lagopus invest more to the development of floral features and to male functions. On the contrary, inbreeding species such as P. patagonica, P. drummondii, and P. ovata invest greater resources to the female function. Received 13 April 1998/ Accepted in revised form 6 November 1998     

Airborne Pollen Grains of Gurdur,Turkey

Ｔｈｃ ｐｏｌｌｅｎ ｇｒａｉｎｓ ｉｎ ｔｈｅ ａｔｍｏｓｐｈｅｒｅ ｏｆ Ｂｕｒｄｕｒ ｈａｖｅ ｂｅｅｎ ｓｔｕｄｉｅｄ ｆｏｒ ａ ｔｗｏ－ｙｅａｒ ｐｅｒｉｏｄ（１９９６ｔｈｒｏｕｇｈ １９９７）ｗｉｔｈ ａ Ｄｕｒｈａｍ ｓａｍｐｌｅｒ．Ａ ｔｏｔａｌ ｏｆ １１ ８８１ ｐｏｌｌｅｎ ｇｒａｉｎｓ ｂｅｌｏｎｇｉｎｓ ｔｏ ３９ ｔａｘａ ｈａｖｅ ｂｅｅｎ ｉｄｅｎｔｉ－ｆｉｅｄ     

Dynamic <Emphasis Type="Italic">in vivo</Emphasis> imaging and cell tracking using a histone fluorescent protein fusion in mice

Background  

Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse transgenic and targeted mutagenesis applications.