Immunofluorescent studies on aphakia,a mutation of a gene involved in the control of lens differentiation in the mouse embryo

Aphakia, an autosomal recessive single gene mutation in the mouse, seriously affects the development of the ocular lens. Up to advanced stages of lens invagination morphogenesis proceeds normally. In the late lens cup and early lens vesicle stage, however, the epithelium of the lens rudiment becomes disorganized and the lumen of the vesicle fills up with rounded cells, apparently released from the epithelium. The lens stalk persists frequently. Probably as a consequence of the aphakic state other parts of the eye secondarily become abnormal.Immunofluorescent studies were done on embryonic normal and aphakia eyes with antisera against adult mouse crystallins. In the normal embryo the first positive reactions were found in the late lens cup stage ($\text{10}\text{3}\text{4}\text{–11}$ days of gestation). By Day 12 all cells of the lens vesicle were brightly fluorescent. A day later the cells of the posterior wall, now lens fibers, had elongated sufficiently to obliterate the lumen of the vesicle. The entire organ was highly fluorescent, indicating that all of its cells contained large amounts of crystallins. The mutant lens, studied over the same time span, showed no reaction at all. The most likely explanation is, that the multiple structural genes, which normally must be involved in the production of the crystallins, are not expressed up to this time in the mutant.The combination of morphological and biochemical defects suggests that the gene involved in the mutation somehow functions in the control of lens differentiation.     

EZYprep LC‐coupled MALDI‐TOF/TOF MS: An improved matrix spray application for phosphopeptide characterisation

The quality of MALDI‐TOF mass spectrometric analysis is highly dependent on the matrix and its deposition strategy. Although different matrix‐deposition methods have specific advantages, one major problem in the field of proteomics, particularly with respect to quantitation, is reproducibility between users or laboratories. Compounding this is the varying crystal homogeneity of matrices depending on the deposition strategy used. Here, we describe a novel optimised matrix‐deposition strategy for LC‐MALDI‐TOF/TOF MS using an automated instrument that produces a nebulised matrix “mist” under controlled atmospheric conditions. Comparisons of this with previously reported strategies showed the method to be advantageous for the atypical matrix, 2,5‐DHB, and improved phosphopeptide ionisation when compared with deposition strategies for CHCA. This optimised DHB matrix‐deposition strategy with LC‐MALDI‐TOF/TOF MS, termed EZYprep LC, was subsequently optimised for phosphoproteome analysis and compared to LC‐ESI‐IT‐MS and a previously reported approach for phosphotyrosine identification and characterisation. These methods were used to map phosphorylation on epidermal growth factor‐stimulated epidermal growth factor receptor to gauge the sensitivity of the proposed method. EZYprep DHB LC‐MALDI‐TOF/TOF MS was able to identify more phosphopeptides and characterise more phosphorylation sites than the other two proteomic strategies, thus proving to be a sensitive approach for phosphoproteome analysis.     

A flyway perspective on food resource abundance in a long-distance migrant, the Eurasian teal (Anas crecca)

Two frequent assumptions about the evolution of long-distance migration in birds are that they travel long distances annually to reach food-rich areas for breeding, and that they time their migratory journey to be at staging sites when the latter provide the best feeding conditions. These assumptions have rarely been properly tested, and there is no study in which a species’ major food types have been measured by standardized methods throughout a flyway and over a large part of the year. We here present such data for Eurasian teal (Anas crecca), converted to a common energetic currency, and collected at wintering, spring staging and breeding sites. Teal did not time migration to maximize local food abundance; most birds left wintering and spring staging sites before a sharp increase in invertebrate food abundance occurred. On the other hand, hatching of ducklings coincided with a peak in invertebrate food abundance on boreal breeding lakes. Mean overall food abundance (invertebrates and seeds combined) did not differ between wintering sites in southern France and breeding sites in northern Sweden at the time of breeding. Our results are inconsistent with the hypothesis that long-distance migration in dabbling ducks has evolved because adult birds gain an immediate pay-off in increased food abundance by flying north in spring. However, our data confirm a selective advantage for breeding at higher latitudes, because hatching of ducklings may coincide with a peak in invertebrate emergence and because longer days may increase the duration of efficient foraging.