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Science:揭示ß-arrestins调节斑马鱼发育机制

摘要 : 美国Durham大学科学家 Wilbanks 等人发现-arrestins通过调控hedgehog信号通路从而调节了斑马鱼的发育。众所周知,-arrestins是典型的七次跨膜蛋白,而以往并不知道它与hedgehog存在关联。这一研究发现,使人类对模式生物zerbfish的发育机制有更清晰的认识。这篇文章发表在本期的Science上。
美国Durham大学科学家Wilbanks等人发现ß-arrestins通过调控hedgehog信号通路从而调节了斑马鱼的发育。众所周知,ß-arrestins是典型的七次跨膜蛋白,而以往并不知道它与hedgehog存在关联。这一研究发现,使人类对模式生物zerbfish的发育机制有更清晰的认识。这篇文章发表在本期的science上。
Science:揭示ß-arrestins调节斑马鱼发育机制
Fig. 1. Western blot analysis of zebrafish ß-arrestin 2 expression. Lysate (500 ng protein) from human embryonic kidney (HEK) 293T cells transiently transfected with either pCDNA3.1(+) vector alone (lane 1) or with zebrafish ß-arrestin 2 cDNA (lane 2). Lysate extracts from zebrafish embryos (48 hours after fertilization, two per lane) that were not injected (lane 3) or were injected with VA-MO (lane 4) or ßarr2-MO (lane 5) were probed with antibody to trout ß-arrestin 2, followed by a secondary horseradish peroxidase–conjugated antibody to rabbit, and then visualized by enhanced chemiluminesence. Equal loading of protein was verified by stripping and reprobing the blot with antibodies to actin. Densitometer quantification of three independent experiments reveals an average of 96 ± 3% reduction in ß-arrestin 2 expression with ßarr-MO.
Science:揭示ß-arrestins调节斑马鱼发育机制
Fig. 2. Phenotypes of ßarr2-MO embryos and Smoothened mutants hi2329b and hi1640 at 24 hours after fertilization. (A to E) Lateral views of whole zebrafish bodies. (F to J) Lateral views of somites, dorsal toward the top, anterior to the right. (K to O) S58 antibody staining of embryos. (P to T) 4D9 antibody staining of embryos.
Science:揭示ß-arrestins调节斑马鱼发育机制
Fig. 3. In situ hybridizations of zebrafish embryos at 27 hours after fertilization for downstream GENEs of the Hh signaling pathway. Embryos were not injected (NI) or were injected, as indicated, with MO to visual cone arrestin or with ß-arrestin 2 mRNA. (A to F) In situ hybridization with nkx2.2 probe. Shh mRNA (100 ng) was injected either alone (D), with VA-MO (E), or with ßarr2-MO (F). Shown are lateral views (dorsal toward the top, anterior to the left). (G to L) Double in situ hybridization with shh (blue stain, white arrows indicating expression at the midline) and ptc (red stain, dark arrows) probes on whole mounts [(G) to (I)] or sectioned embryos [(J) to (L)].
Science:揭示ß-arrestins调节斑马鱼发育机制
Fig. 4. Loss of slow muscle fiber development in ßarr2-MO is restored with DNPKA mRNA or Su(fu)-MO. quantitative analysis of slow muscle fiber number per somite in embryos 24 hours after fertilization, injected as indicated. Embryos were stained with S58 antibody. Slow muscle fibers in three to five somites on 20 embryos per experiment were counted. 全文见:http://www.sciencemag.org/cgi/content/full/306/5705/2264 作者:网友 点击:
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