Boylan KL , Mische S , Li M , Marqués G , Morin X , Chia W , Hays TS .

GM53695 NIGMS NIH HHS , R01 GM44757 NIGMS NIH HHS , R21 NS051319 NINDS NIH HHS , Wellcome Trust , R01 GM053695 NIGMS NIH HHS , R01 GM044757 NIGMS NIH HHS

	Figure 1. GFP-Imp Is Expressed in Ovaries and Neurons(A) In the ovary, GFP-Imp is detected within the germarium as early as stage 1 (arrow) and accumulates in the presumptive oocyte in early egg chambers. By stage 9–10, GFP-Imp is concentrated at the oocyte posterior pole (arrowheads). Bar, 25 μm.(B) GFP-Imp is enriched in the embryonic central nervous system (Bar, 25μm), and (C) in larval segmental axons and at the synapse (arrowhead). Bar, 5μm.(D) Schematic highlights the domain structure of Drosophila Imp compared to orthologs in human, chick, and Xenopus. RRM = RNA recognition motif. KH = hnRNP K homology domain.(E) The insertion of the PTT does not alter the level of Imp protein expression. Ovary extracts from wild type and GFP-Imp flies show similar levels of overall expression, as well as similar relative expression in supernatant (S) and pellet (P) fractions, as detected by an anti-Imp antibody.(F) GFP-Imp is present in a large, RNase-sensitive complex. GFP-Imp ovary extracts were fractionated over a 10%–40% sucrose gradient in the presence of RNase, or RNase inhibitors. Equal volumes of alternate fractions were analyzed by western blot using anti-GFP antibody. Lane L, sample of extract loaded on gradient. Fraction numbers are indicated above the lanes (bottom of gradient is fraction 1).
	Figure 2. GFP-Imp Localization in the Oocyte Requires Microtubule Motor Function(A, A') GFP-Imp expression in wild type egg chambers is enriched in the oocyte and localizes to the posterior of the oocyte in stage 10.(B, B') Disruption of dynein using ΔGl overexpression (or the Dhc 6–6/6–12 transheterozygous mutant background; data not shown) produces an early aberrant accumulation of GFP-Imp to the oocyte anterior. However, the subsequent accumulation to the posterior pole is not affected.(C, C') In germline clones homozygous for the kinesin null mutation, khc27, GFP-Imp enrichment in the early oocyte is retained, but later accumulation at the posterior of the oocyte is lost. Bar (applies to all images), 10 μm.
	Figure 3. Dynein and Kinesin Have Opposing Effects on GFP-Imp Motility in Nurse Cells(A) Three sequential images are separately pseudocolored red, green, or blue, and overlaid to highlight the frequency of GFP-Imp particle movements in the nurse cell cytoplasm in wild type and mutant backgrounds. Motility is easily discerned in wild type by the resolution of individual colors. After disruption of dynein (Dhc-) or dynactin (ΔGl), there is almost no movement; particles appear white, resulting from the superimposition of red, green, and blue. In contrast, disruption of kinesin I (Khc-) increases particle transport, as demonstrated by the increased visibility of individual colors. Bar, 10 μm. Genetic backgrounds shown include Dhc-, [Dhc6–6/Dhc6–12]; ΔGl, [UASp-ΔGl/+; nanos-GAL4/+]; and khc27, [khc27/ khc27 ] germline clones, as described in the text.(B) Motor mutations affect velocity as well as frequency of GFP-Imp transport events. Histogram illustrates the average velocities of GFP-Imp particles in nurse cells derived from wild type and mutant backgrounds. (1) wild type, (2) Dhc6–6/Dhc6–12, (3) UASp-ΔGl/+; nanos-GAL4, (4) khc27/ khc27germline clone, (5) wild type treated with colcemid, (6) wild type treated with cytochalasin D.
	Figure 4. Imp Expression Affects Synaptic Terminal Size at the Neuromuscular Junction(A) Individual synaptic boutons are clearly labeled by the anti-CSP antibody. Wild type larval synapse is shown.(B) Bouton numbers are decreased in the Imp mutant combination, H44/H149.(C) Presynaptic overexpression of the UAS-RE transgene by elav-GAL4 (elav-GAL4/+; UAS-RE/+) significantly increases the number of boutons. Bar (applies to A–C), 50 μm.(D) Histogram compares relative synaptic terminal sizes of (1) wild type, (2) Imp H44/H149 mutant combination, and (3–5) overexpression of Imp transgenes, including (3) elav-GAL4/+; Imp-RE, (4) elav-GAL4/+; Imp-SD/+, and (5) elav-GAL4/+; Imp-KH/+. The synaptic terminal size was defined as the number of synaptic boutons normalized to the surface area of muscle 6/7, and the reference was set to 100 for the wild type control.(E) GFP-Imp is present at the synapse and accumulates along the membrane cortex of synaptic boutons (arrow). Bar, 5 μm.
	Figure 5. Imp Mutations Do Not Disrupt the Nuclear Accumulation of pMad(A) In wild type, pMad is readily detected in the nuclei of motorneuron ganglia.(B) Nuclear uptake of pMad is retained in the Imp transheterozygous H44/H149 mutant background.(C) In contrast, pMad staining is abolished in the wit mutant. Bar, 10 μm.

Aberle, wishful thinking encodes a BMP type II receptor that regulates synaptic growth in Drosophila. 2002, Pubmed

Aberle, wishful thinking encodes a BMP type II receptor that regulates synaptic growth in Drosophila. 2002, Pubmed
Ainger, Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes. 1993, Pubmed
Barbee, Staufen- and FMRP-containing neuronal RNPs are structurally and functionally related to somatic P bodies. 2006, Pubmed
Boylan, The gene for the intermediate chain subunit of cytoplasmic dynein is essential in Drosophila. 2002, Pubmed , Xenbase
Brendza, Clonal tests of conventional kinesin function during cell proliferation and differentiation. 2000, Pubmed
Brendza, A function for kinesin I in the posterior transport of oskar mRNA and Staufen protein. 2000, Pubmed
Bullock, Conserved signals and machinery for RNA transport in Drosophila oogenesis and embryogenesis. 2001, Pubmed
Bullock, Guidance of bidirectional motor complexes by mRNA cargoes through control of dynein number and activity. 2006, Pubmed
Cha, In vivo analysis of Drosophila bicoid mRNA localization reveals a novel microtubule-dependent axis specification pathway. 2001, Pubmed
Cooperstock, RNA localization and translational regulation during axis specification in the Drosophila oocyte. 2001, Pubmed
Czaplinski, Pathways for mRNA localization in the cytoplasm. 2006, Pubmed
Dermaut, Aberrant lysosomal carbohydrate storage accompanies endocytic defects and neurodegeneration in Drosophila benchwarmer. 2005, Pubmed
Deshler, Localization of Xenopus Vg1 mRNA by Vera protein and the endoplasmic reticulum. 1997, Pubmed , Xenbase
Farina, Two ZBP1 KH domains facilitate beta-actin mRNA localization, granule formation, and cytoskeletal attachment. 2003, Pubmed
Feng, A modified minimal hemolymph-like solution, HL3.1, for physiological recordings at the neuromuscular junctions of normal and mutant Drosophila larvae. 2004, Pubmed
Gamberi, Drosophila RNA binding proteins. 2006, Pubmed
Geng, Imp associates with squid and Hrp48 and contributes to localized expression of gurken in the oocyte. 2006, Pubmed
Gepner, Cytoplasmic dynein function is essential in Drosophila melanogaster. 1996, Pubmed
Golby, Partitioning of N-ethylmaleimide-sensitive fusion (NSF) protein function in Drosophila melanogaster: dNSF1 is required in the nervous system, and dNSF2 is required in mesoderm. 2001, Pubmed
Havin, RNA-binding protein conserved in both microtubule- and microfilament-based RNA localization. 1998, Pubmed , Xenbase
Hüttelmaier, Spatial regulation of beta-actin translation by Src-dependent phosphorylation of ZBP1. 2005, Pubmed
Jambhekar, Cis-acting determinants of asymmetric, cytoplasmic RNA transport. 2007, Pubmed
Karess, Analysis of P transposable element functions in Drosophila. 1984, Pubmed
Kislauskis, Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype. 1994, Pubmed
Koch, Multiple effects of colchicine on oogenesis in Drosophila: induced sterility and switch of potential oocyte to nurse-cell developmental pathway. 1983, Pubmed
Korey, small bristles is required for the morphogenesis of multiple tissues during Drosophila development. 2001, Pubmed
Kraut, A gain-of-function screen for genes controlling motor axon guidance and synaptogenesis in Drosophila. 2001, Pubmed
Leeds, Developmental regulation of CRD-BP, an RNA-binding protein that stabilizes c-myc mRNA in vitro. 1997, Pubmed
Leung, Asymmetrical beta-actin mRNA translation in growth cones mediates attractive turning to netrin-1. 2006, Pubmed , Xenbase
Lipshitz, Mechanisms of RNA localization and translational regulation. 2000, Pubmed
Litman, Subcellular localization of tau mRNA in differentiating neuronal cell culture: implications for neuronal polarity. 1993, Pubmed
MacDougall, Drosophila gurken (TGFalpha) mRNA localizes as particles that move within the oocyte in two dynein-dependent steps. 2003, Pubmed
Marqués, The Drosophila BMP type II receptor Wishful Thinking regulates neuromuscular synapse morphology and function. 2002, Pubmed
McCabe, Highwire regulates presynaptic BMP signaling essential for synaptic growth. 2004, Pubmed
McCabe, The BMP homolog Gbb provides a retrograde signal that regulates synaptic growth at the Drosophila neuromuscular junction. 2003, Pubmed
McGrail, The microtubule motor cytoplasmic dynein is required for spindle orientation during germline cell divisions and oocyte differentiation in Drosophila. 1997, Pubmed
Mische, Direct observation of regulated ribonucleoprotein transport across the nurse cell/oocyte boundary. 2007, Pubmed
Morales, Drosophila fragile X protein, DFXR, regulates neuronal morphology and function in the brain. 2002, Pubmed
Morin, A protein trap strategy to detect GFP-tagged proteins expressed from their endogenous loci in Drosophila. 2001, Pubmed
Munro, A repeated IMP-binding motif controls oskar mRNA translation and anchoring independently of Drosophila melanogaster IMP. 2006, Pubmed , Xenbase
Nielsen, Cytoplasmic trafficking of IGF-II mRNA-binding protein by conserved KH domains. 2002, Pubmed
Nielsen, A family of insulin-like growth factor II mRNA-binding proteins represses translation in late development. 1999, Pubmed , Xenbase
Nielsen, The biphasic expression of IMP/Vg1-RBP is conserved between vertebrates and Drosophila. 2000, Pubmed , Xenbase
Norga, Quantitative analysis of bristle number in Drosophila mutants identifies genes involved in neural development. 2003, Pubmed
Norvell, Specific isoforms of squid, a Drosophila hnRNP, perform distinct roles in Gurken localization during oogenesis. 1999, Pubmed
Noubissi, CRD-BP mediates stabilization of betaTrCP1 and c-myc mRNA in response to beta-catenin signalling. 2006, Pubmed
Oberman, VICKZ proteins mediate cell migration via their RNA binding activity. 2007, Pubmed , Xenbase
Oleynikov, Real-time visualization of ZBP1 association with beta-actin mRNA during transcription and localization. 2003, Pubmed
Palacios, Getting the message across: the intracellular localization of mRNAs in higher eukaryotes. 2001, Pubmed
Persson, The L45 loop in type I receptors for TGF-beta family members is a critical determinant in specifying Smad isoform activation. 1998, Pubmed , Xenbase
Pokrywka, Microtubules are a general component of mRNA localization systems in Drosophila oocytes. 1995, Pubmed
Quiñones-Coello, Exploring strategies for protein trapping in Drosophila. 2007, Pubmed
Rawson, Drosophila neuromuscular synapse assembly and function require the TGF-beta type I receptor saxophone and the transcription factor Mad. 2003, Pubmed
Rørth, Gal4 in the Drosophila female germline. 1998, Pubmed
Ross, Characterization of a beta-actin mRNA zipcode-binding protein. 1997, Pubmed
Sossin, Intracellular trafficking of RNA in neurons. 2006, Pubmed
Steinhauer, Microtubule polarity and axis formation in the Drosophila oocyte. 2006, Pubmed
Steward, A cellular mechanism for targeting newly synthesized mRNAs to synaptic sites on dendrites. 2001, Pubmed
Steward, Synaptic activation causes the mRNA for the IEG Arc to localize selectively near activated postsynaptic sites on dendrites. 1998, Pubmed
Stewart, Population density regulates Drosophila synaptic morphology in a Fasciclin-II-dependent manner. 2004, Pubmed
St Johnston, Moving messages: the intracellular localization of mRNAs. 2005, Pubmed
Sundell, Requirement of microfilaments in sorting of actin messenger RNA. 1991, Pubmed
Sweeney, Unrestricted synaptic growth in spinster-a late endosomal protein implicated in TGF-beta-mediated synaptic growth regulation. 2002, Pubmed
Theurkauf, A central role for microtubules in the differentiation of Drosophila oocytes. 1993, Pubmed
Vikesaa, RNA-binding IMPs promote cell adhesion and invadopodia formation. 2006, Pubmed
Vilinsky, A Drosophila SNAP-25 null mutant reveals context-dependent redundancy with SNAP-24 in neurotransmission. 2002, Pubmed
Wan, Characterization of dFMR1, a Drosophila melanogaster homolog of the fragile X mental retardation protein. 2000, Pubmed
Wan, Highwire regulates synaptic growth in Drosophila. 2000, Pubmed
Wang, Identification and testing of a gene expression signature of invasive carcinoma cells within primary mammary tumors. 2004, Pubmed
Wanner, Changing subcellular distribution and activity-dependent utilization of a dendritically localized mRNA in developing Purkinje cells. 2000, Pubmed
Wilhelm, Isolation of a ribonucleoprotein complex involved in mRNA localization in Drosophila oocytes. 2000, Pubmed
Wilkie, Drosophila wingless and pair-rule transcripts localize apically by dynein-mediated transport of RNA particles. 2001, Pubmed
Yao, Actin-dependent activation of presynaptic silent synapses contributes to long-term synaptic plasticity in developing hippocampal neurons. 2006, Pubmed
Yisraeli, VICKZ proteins: a multi-talented family of regulatory RNA-binding proteins. 2005, Pubmed
Zarnescu, Fragile X protein functions with lgl and the par complex in flies and mice. 2005, Pubmed
Zhang, Drosophila fragile X-related gene regulates the MAP1B homolog Futsch to control synaptic structure and function. 2001, Pubmed
Zhang, stress sensitive B encodes an adenine nucleotide translocase in Drosophila melanogaster. 1999, Pubmed