As in the last stripes to become resolved are stripes 4 and 5, whose inter-stripe domain clears only around T7. profiles used for data quantification, per gene and time class. 2041-9139-5-1-S2.pdf Ononin (56K) GUID:?3A480E49-3CF5-49E1-8D97-E090AB2D9F3B Additional file 3: Table S3 Number of embryos per time class according to physiological age. This table plots the number of embryos per assigned time class (C10-C13, C14A: T1-T8) versus the time of fixation (in hours:minutes after egg activation). See Methods in the main text for details on egg activation, embryo fixation, and assignment of embryos to time classes. 2041-9139-5-1-S3.pdf (77K) GUID:?3E130354-BCD0-45C0-8EDC-7CC4517C0C8A Additional file 4: Table S4 Average positions and shifts of Eve stripe peaks in ((and embryos. Data shown for time classes T5-T8. Total domain widths (in % egg length) are calculated – from integrated data – as reaching from the peak of Eve stripe 1 to the peak of Eve stripe 6 (Our data reveal differences in the dynamics of Hb boundary positioning and Eve stripe formation between and and provide a starting point for comparative reverse-engineering studies of the evolutionary and developmental dynamics of the segmentation gene system. (family: Psychodidae; Figure?1). Gap and pair-rule gene expression in this species shows a number of interesting qualitative differences compared to the standard model for dipteran segmentation, the vinegar fly In particular, expression of posterior gap domains is modified and pair-rule stripes are delayed in compared to (Figure?1; [52-54]). To investigate these differences in more detail, we use immunofluorescence combined with confocal scanning microscopy and a data processing pipeline adapted from We analyze these expression patterns with regard to their timing and spatial registration. Our results show that the dynamics of Hb boundary positioning and Eve stripe formation differ significantly between and Anterior shifts in domain position as development proceeds are not only present, but much more pronounced in Despite this, the relative arrangement of the anterior Hb domain and the anterior stripes of Eve is largely conserved across the evolutionary distance between the two species. The quantitative dataset that we have produced provides a suitable starting point for future reverse-engineering approaches to study the mechanisms and dynamics of segmentation gene regulation in (family: Culicidae) and (Psychodidae) both belong to the paraphyletic assemblage of the Nematocera. (Drosophilidae) and (Syrphidae) belong to the monophyletic cyclorrhaphan Brachycera (higher flies). The column of graphs on the Ononin left CCL2 schematically shows the presence and expression patterns of maternal co-ordinate genes; graphs on the right show zygotically expressed gap gene domains (at the end of C14A, just before gastrulation; solid triangles), and the delayed appearance of the posterior Hb domain in (after gastrulation; triangle with dashed outline). X-axes represent the antero-posterior embryonic axis: anterior is to the left, posterior to the right. Maternal co-ordinate gene products: Bcd, Bicoid; Hb, Hunchback; Cad, Caudal. Gap gene products: Hb, Hunchback; Kr, Krppel; Kni, Knirps; Gt, Giant; Tll, Tailless; Hkb, Huckebein. Fly images from the Encyclopedia of Ononin Life (http://www.eol.org), photographers: culture and embryo collection methods have been described previously [54] (detailed protocols are available from the authors on demand). Embryos had been dissected from adult females, advancement was turned on by osmotic surprise, and embryos had been still left to develop before preferred stage on damp filtration system paper at 25C. Blastoderm stage embryos had been fixed at several period points of advancement up to optimum of 8 hours after egg activation (find Results for information). Fixation was performed utilizing a adjustment of the described method [54] previously. Pursuing dechorionation in 50% bleach, and fixation within a 1:1 combination of 8% formaldehyde in PBS/heptane for 25 a few minutes, the formaldehyde/heptane was taken out. Heptane was put into the embryos accompanied by an equal level of methanol (pre-cooled to ?80C), which mix was shaken for 20 to 30 secs to fracture the vitelline layer. The heptane/methanol was replaced and removed with methanol. Embryos were used in a 10-ml syringe (installed using a.
