Shipp Lauren E, Hill Rose Z, Moy Gary W, Gökırmak Tufan, Hamdoun Amro
Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0202, USA.
Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0202, USA
Development. 2015 Oct 15;142(20):3537-48. doi: 10.1242/dev.126144. Epub 2015 Sep 22.
ATP-binding cassette (ABC) transporters are evolutionarily conserved proteins that pump diverse substrates across membranes. Many are known to efflux signaling molecules and are extensively expressed during development. However, the role of transporters in moving extracellular signals that regulate embryogenesis is largely unexplored. Here, we show that a mesodermal ABCC (MRP) transporter is necessary for endodermal gut morphogenesis in sea urchin embryos. This transporter, Sp-ABCC5a (C5a), is expressed in pigment cells and their precursors, which are a subset of the non-skeletogenic mesoderm (NSM) cells. C5a expression depends on Delta/Notch signaling from skeletogenic mesoderm and is downstream of Gcm in the aboral NSM gene regulatory network. Long-term imaging of development reveals that C5a knockdown embryos gastrulate, but ∼90% develop a prolapse of the hindgut by the late prism stage (∼8 h after C5a protein expression normally peaks). Since C5a orthologs efflux cyclic nucleotides, and cAMP-dependent protein kinase (Sp-CAPK/PKA) is expressed in pigment cells, we examined whether C5a could be involved in gastrulation through cAMP transport. Consistent with this hypothesis, membrane-permeable pCPT-cAMP rescues the prolapse phenotype in C5a knockdown embryos, and causes archenteron hyper-invagination in control embryos. In addition, the cAMP-producing enzyme soluble adenylyl cyclase (sAC) is expressed in pigment cells, and its inhibition impairs gastrulation. Together, our data support a model in which C5a transports sAC-derived cAMP from pigment cells to control late invagination of the hindgut. Little is known about the ancestral functions of ABCC5/MRP5 transporters, and this study reveals a novel role for these proteins in mesoderm-endoderm signaling during embryogenesis.
ATP结合盒(ABC)转运蛋白是进化上保守的蛋白质,可将多种底物泵过细胞膜。已知许多ABC转运蛋白能外排信号分子,并在发育过程中广泛表达。然而,转运蛋白在传递调节胚胎发生的细胞外信号中的作用在很大程度上尚未被探索。在这里,我们表明中胚层ABCC(多药耐药相关蛋白,MRP)转运蛋白对于海胆胚胎内胚层肠道形态发生是必需的。这种转运蛋白,即Sp-ABCC5a(C5a),在色素细胞及其前体中表达,色素细胞及其前体是非造骨中胚层(NSM)细胞的一个子集。C5a的表达依赖于来自造骨中胚层的Delta/Notch信号,并且在口面NSM基因调控网络中位于Gcm的下游。发育的长期成像显示,C5a基因敲低的胚胎会进行原肠胚形成,但约90%的胚胎在晚期棱柱期(C5a蛋白表达通常达到峰值后约8小时)会出现后肠脱垂。由于C5a直系同源物能外排环核苷酸,并且环磷酸腺苷依赖性蛋白激酶(Sp-CAPK/PKA)在色素细胞中表达,我们研究了C5a是否可能通过环磷酸腺苷转运参与原肠胚形成。与这一假设一致,膜通透性的pCPT-环磷酸腺苷可挽救C5a基因敲低胚胎中的脱垂表型,并在对照胚胎中导致原肠过度内陷。此外,产生环磷酸腺苷的酶可溶性腺苷酸环化酶(sAC)在色素细胞中表达,其抑制会损害原肠胚形成。总之,我们的数据支持一个模型,即C5a将sAC衍生的环磷酸腺苷从色素细胞转运出来,以控制后肠的晚期内陷。关于ABCC5/MRP5转运蛋白祖先功能的了解很少,而这项研究揭示了这些蛋白质在胚胎发生过程中中胚层-内胚层信号传导中的新作用。