Abstract
R8 photoreceptor cells play a primary role in the differentiation of Drosophila eyes. In scabrous (sca) mutants, the pattern of R8 photoreceptor differentiation is altered. The sca gene is predicted to encode a secreted protein related in part to fibrinogen and tenascins. Using expression in Drosophila Schneider cells, we showed that sca encoded a dimeric glycoprotein which was secreted and found in soluble form in the tissue culture medium. The sca protein contained both N- and O-linked carbohydrates and interacted with heparin. This Schneider cell protein was similar to protein detected in embryos. We showed that sca mutations, along with conditional alleles of Notch (N) and Delta (Dl), each affected the pattern of cells expressing atonal (ato), the proneural gene required for R8 differentiation. In normal development, about 1 cell in 20 differentiates into an R8 cell; in the others, ato is repressed. N and Dl were required to repress ato in the vicinity of R8 cells, whereas sca had effects over several cell diameters. Certain antibodies detected uptake of sca protein several cells away from its source. The overall growth factor-like structure of sca protein, its solubility, and its range of effects in vivo are consistent with a diffusible role that complements mechanisms involving direct cell contact. We propose that as the morphogenic furrow advances, cell secreting sca protein control the pattern of the next ommatidial column.
Full Text
The Full Text of this article is available as a PDF (1.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Artavanis-Tsakonas S., Matsuno K., Fortini M. E. Notch signaling. Science. 1995 Apr 14;268(5208):225–232. doi: 10.1126/science.7716513. [DOI] [PubMed] [Google Scholar]
- Ashburner M., Bonner J. J. The induction of gene activity in drosophilia by heat shock. Cell. 1979 Jun;17(2):241–254. doi: 10.1016/0092-8674(79)90150-8. [DOI] [PubMed] [Google Scholar]
- Baker N. E., Mlodzik M., Rubin G. M. Spacing differentiation in the developing Drosophila eye: a fibrinogen-related lateral inhibitor encoded by scabrous. Science. 1990 Dec 7;250(4986):1370–1377. doi: 10.1126/science.2175046. [DOI] [PubMed] [Google Scholar]
- Baker N. E., Rubin G. M. Ellipse mutations in the Drosophila homologue of the EGF receptor affect pattern formation, cell division, and cell death in eye imaginal discs. Dev Biol. 1992 Apr;150(2):381–396. doi: 10.1016/0012-1606(92)90250-k. [DOI] [PubMed] [Google Scholar]
- Baker N. E., Zitron A. E. Drosophila eye development: Notch and Delta amplify a neurogenic pattern conferred on the morphogenetic furrow by scabrous. Mech Dev. 1995 Feb;49(3):173–189. doi: 10.1016/0925-4773(94)00314-d. [DOI] [PubMed] [Google Scholar]
- Bernfield M., Kokenyesi R., Kato M., Hinkes M. T., Spring J., Gallo R. L., Lose E. J. Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol. 1992;8:365–393. doi: 10.1146/annurev.cb.08.110192.002053. [DOI] [PubMed] [Google Scholar]
- Bradley R. S., Brown A. M. The proto-oncogene int-1 encodes a secreted protein associated with the extracellular matrix. EMBO J. 1990 May;9(5):1569–1575. doi: 10.1002/j.1460-2075.1990.tb08276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown N. L., Sattler C. A., Paddock S. W., Carroll S. B. Hairy and emc negatively regulate morphogenetic furrow progression in the Drosophila eye. Cell. 1995 Mar 24;80(6):879–887. doi: 10.1016/0092-8674(95)90291-0. [DOI] [PubMed] [Google Scholar]
- Bunch T. A., Grinblat Y., Goldstein L. S. Characterization and use of the Drosophila metallothionein promoter in cultured Drosophila melanogaster cells. Nucleic Acids Res. 1988 Feb 11;16(3):1043–1061. doi: 10.1093/nar/16.3.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cubas P., de Celis J. F., Campuzano S., Modolell J. Proneural clusters of achaete-scute expression and the generation of sensory organs in the Drosophila imaginal wing disc. Genes Dev. 1991 Jun;5(6):996–1008. doi: 10.1101/gad.5.6.996. [DOI] [PubMed] [Google Scholar]
- Doe C. Q., Goodman C. S. Early events in insect neurogenesis. II. The role of cell interactions and cell lineage in the determination of neuronal precursor cells. Dev Biol. 1985 Sep;111(1):206–219. doi: 10.1016/0012-1606(85)90446-4. [DOI] [PubMed] [Google Scholar]
- Doolittle R. F. A detailed consideration of a principal domain of vertebrate fibrinogen and its relatives. Protein Sci. 1992 Dec;1(12):1563–1577. doi: 10.1002/pro.5560011204. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edge A. S., Faltynek C. R., Hof L., Reichert L. E., Jr, Weber P. Deglycosylation of glycoproteins by trifluoromethanesulfonic acid. Anal Biochem. 1981 Nov 15;118(1):131–137. doi: 10.1016/0003-2697(81)90168-8. [DOI] [PubMed] [Google Scholar]
- Ellis M. C., Weber U., Wiersdorff V., Mlodzik M. Confrontation of scabrous expressing and non-expressing cells is essential for normal ommatidial spacing in the Drosophila eye. Development. 1994 Jul;120(7):1959–1969. doi: 10.1242/dev.120.7.1959. [DOI] [PubMed] [Google Scholar]
- Erickson H. P. Tenascin-C, tenascin-R and tenascin-X: a family of talented proteins in search of functions. Curr Opin Cell Biol. 1993 Oct;5(5):869–876. doi: 10.1016/0955-0674(93)90037-q. [DOI] [PubMed] [Google Scholar]
- Fradkin L. G., Noordermeer J. N., Nusse R. The Drosophila Wnt protein DWnt-3 is a secreted glycoprotein localized on the axon tracts of the embryonic CNS. Dev Biol. 1995 Mar;168(1):202–213. doi: 10.1006/dbio.1995.1072. [DOI] [PubMed] [Google Scholar]
- Hager D. A., Burgess R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 1980 Nov 15;109(1):76–86. doi: 10.1016/0003-2697(80)90013-5. [DOI] [PubMed] [Google Scholar]
- Heitzler P., Simpson P. Altered epidermal growth factor-like sequences provide evidence for a role of Notch as a receptor in cell fate decisions. Development. 1993 Mar;117(3):1113–1123. doi: 10.1242/dev.117.3.1113. [DOI] [PubMed] [Google Scholar]
- Heitzler P., Simpson P. The choice of cell fate in the epidermis of Drosophila. Cell. 1991 Mar 22;64(6):1083–1092. doi: 10.1016/0092-8674(91)90263-x. [DOI] [PubMed] [Google Scholar]
- Hu X., Lee E. C., Baker N. E. Molecular analysis of scabrous mutant alleles from Drosophila melanogaster indicates a secreted protein with two functional domains. Genetics. 1995 Oct;141(2):607–617. doi: 10.1093/genetics/141.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jarman A. P., Grell E. H., Ackerman L., Jan L. Y., Jan Y. N. Atonal is the proneural gene for Drosophila photoreceptors. Nature. 1994 Jun 2;369(6479):398–400. doi: 10.1038/369398a0. [DOI] [PubMed] [Google Scholar]
- Jarman A. P., Sun Y., Jan L. Y., Jan Y. N. Role of the proneural gene, atonal, in formation of Drosophila chordotonal organs and photoreceptors. Development. 1995 Jul;121(7):2019–2030. doi: 10.1242/dev.121.7.2019. [DOI] [PubMed] [Google Scholar]
- Jokerst R. S., Weeks J. R., Zehring W. A., Greenleaf A. L. Analysis of the gene encoding the largest subunit of RNA polymerase II in Drosophila. Mol Gen Genet. 1989 Jan;215(2):266–275. doi: 10.1007/BF00339727. [DOI] [PubMed] [Google Scholar]
- Kiefer P., Peters G., Dickson C. The Int-2/Fgf-3 oncogene product is secreted and associates with extracellular matrix: implications for cell transformation. Mol Cell Biol. 1991 Dec;11(12):5929–5936. doi: 10.1128/mcb.11.12.5929. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lindquist S. Regulation of protein synthesis during heat shock. Nature. 1981 Sep 24;293(5830):311–314. doi: 10.1038/293311a0. [DOI] [PubMed] [Google Scholar]
- Mlodzik M., Baker N. E., Rubin G. M. Isolation and expression of scabrous, a gene regulating neurogenesis in Drosophila. Genes Dev. 1990 Nov;4(11):1848–1861. doi: 10.1101/gad.4.11.1848. [DOI] [PubMed] [Google Scholar]
- Panganiban G. E., Rashka K. E., Neitzel M. D., Hoffmann F. M. Biochemical characterization of the Drosophila dpp protein, a member of the transforming growth factor beta family of growth factors. Mol Cell Biol. 1990 Jun;10(6):2669–2677. doi: 10.1128/mcb.10.6.2669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Papkoff J., Schryver B. Secreted int-1 protein is associated with the cell surface. Mol Cell Biol. 1990 Jun;10(6):2723–2730. doi: 10.1128/mcb.10.6.2723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parks A. L., Turner F. R., Muskavitch M. A. Relationships between complex Delta expression and the specification of retinal cell fates during Drosophila eye development. Mech Dev. 1995 Apr;50(2-3):201–216. doi: 10.1016/0925-4773(94)00336-l. [DOI] [PubMed] [Google Scholar]
- Ready D. F., Hanson T. E., Benzer S. Development of the Drosophila retina, a neurocrystalline lattice. Dev Biol. 1976 Oct 15;53(2):217–240. doi: 10.1016/0012-1606(76)90225-6. [DOI] [PubMed] [Google Scholar]
- Rettenmier C. W., Roussel M. F., Ashmun R. A., Ralph P., Price K., Sherr C. J. Synthesis of membrane-bound colony-stimulating factor 1 (CSF-1) and downmodulation of CSF-1 receptors in NIH 3T3 cells transformed by cotransfection of the human CSF-1 and c-fms (CSF-1 receptor) genes. Mol Cell Biol. 1987 Jul;7(7):2378–2387. doi: 10.1128/mcb.7.7.2378. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rio D. C., Rubin G. M. Transformation of cultured Drosophila melanogaster cells with a dominant selectable marker. Mol Cell Biol. 1985 Aug;5(8):1833–1838. doi: 10.1128/mcb.5.8.1833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schweitzer R., Shaharabany M., Seger R., Shilo B. Z. Secreted Spitz triggers the DER signaling pathway and is a limiting component in embryonic ventral ectoderm determination. Genes Dev. 1995 Jun 15;9(12):1518–1529. doi: 10.1101/gad.9.12.1518. [DOI] [PubMed] [Google Scholar]
- Simon M. A., Bowtell D. D., Rubin G. M. Structure and activity of the sevenless protein: a protein tyrosine kinase receptor required for photoreceptor development in Drosophila. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8333–8337. doi: 10.1073/pnas.86.21.8333. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simpson P. Lateral inhibition and the development of the sensory bristles of the adult peripheral nervous system of Drosophila. Development. 1990 Jul;109(3):509–519. doi: 10.1242/dev.109.3.509. [DOI] [PubMed] [Google Scholar]
- Simpson P. Notch and the choice of cell fate in Drosophila neuroepithelium. Trends Genet. 1990 Nov;6(11):343–345. doi: 10.1016/0168-9525(90)90260-d. [DOI] [PubMed] [Google Scholar]
- Singson A., Leviten M. W., Bang A. G., Hua X. H., Posakony J. W. Direct downstream targets of proneural activators in the imaginal disc include genes involved in lateral inhibitory signaling. Genes Dev. 1994 Sep 1;8(17):2058–2071. doi: 10.1101/gad.8.17.2058. [DOI] [PubMed] [Google Scholar]
- Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
- Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
- Tomlinson A., Ready D. F. Neuronal differentiation in Drosophila ommatidium. Dev Biol. 1987 Apr;120(2):366–376. doi: 10.1016/0012-1606(87)90239-9. [DOI] [PubMed] [Google Scholar]
- Vincent J. P., Lawrence P. A. Drosophila wingless sustains engrailed expression only in adjoining cells: evidence from mosaic embryos. Cell. 1994 Jun 17;77(6):909–915. doi: 10.1016/0092-8674(94)90139-2. [DOI] [PubMed] [Google Scholar]
- Wolff T., Ready D. F. The beginning of pattern formation in the Drosophila compound eye: the morphogenetic furrow and the second mitotic wave. Development. 1991 Nov;113(3):841–850. doi: 10.1242/dev.113.3.841. [DOI] [PubMed] [Google Scholar]
- Yost H. J., Lindquist S. RNA splicing is interrupted by heat shock and is rescued by heat shock protein synthesis. Cell. 1986 Apr 25;45(2):185–193. doi: 10.1016/0092-8674(86)90382-x. [DOI] [PubMed] [Google Scholar]
- Zipursky S. L., Rubin G. M. Determination of neuronal cell fate: lessons from the R7 neuron of Drosophila. Annu Rev Neurosci. 1994;17:373–397. doi: 10.1146/annurev.ne.17.030194.002105. [DOI] [PubMed] [Google Scholar]
- van den Heuvel M., Harryman-Samos C., Klingensmith J., Perrimon N., Nusse R. Mutations in the segment polarity genes wingless and porcupine impair secretion of the wingless protein. EMBO J. 1993 Dec 15;12(13):5293–5302. doi: 10.1002/j.1460-2075.1993.tb06225.x. [DOI] [PMC free article] [PubMed] [Google Scholar]