Abstract
Using the conserved basic helix-loop-helix (bHLH) domain of AS-C and MASH, we amplified by polymerase chain reaction, the HLH region of a chick AS-C/MASH homologue from cDNA prepared from poly (A+) RNA isolated from a region rich in cardiac neural crest cells. A full-length cDNA clone was obtained from a stage 23 chick head cDNA library using this conserved sequence as a probe. Comparison of this clone with MASH1 revealed strong homologies in the bHLH domain and carboxyl terminal region of the conceptually translated protein. The clone was named chick achaete-scute homologue (CASH). This transcript has a single open reading frame that codes for a protein of 218 residues. The deduced amino acid sequence of CASH is 100% identical to MASH1 in the bHLH region and 83.9% identical over the entire protein. The bHLH domain of CASH is highly conserved within the AS-C family. Northern analysis indicates that CASH is expressed transiently in a defined spatial pattern. The highest expression is in the head between stages 23 and 26. Whole mount in situ hybridization suggests that CASH is expressed at stage 23 in the dorsal ventricular zone and central sensory pathways and later in the ventral ventricular zone and ventral horn. By stage 28, it is expressed ubiquitously in the peripheral nervous system including all of the neurons derived from the the neural crest and epipharyngeal placodes, as well as neural crest-derived chromaffin cells in the adrenal medulla. Amplification by polymerase chain reaction indicates that CASH is expressed in the cardiac neural fold and continues to be expressed in cultured cardiac neural crest cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdulla R-I, Slott EF, Kirby ML (1993) Proteins associated with cardiac neural crest in the pharyngeal region of early chick embryos. Pediatr Res 33:43–47
Alonso MC, Cabrera CV (1988) The achaete-scute complex of Drosophila melanogaster comprises four homologus genes. EMBO J 7:2585–2591
Benezra R, Davis RL, Lockshou D, Turner DL, Weintraub H (1990) The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell 61:49–59
Cabrera CV, Martínez-Arias A, Bate M (1987) The expression of three members of the achaete-scute gene complex correlates with neuroblast segregation in Drosophila. Cell 50:202–208
Caudy M, Grell EH, Dambly-Chaudière G, Ghysen A, Jan LY, Jan YN (1988a) The maternal sex determination gene daughterless has has zygotic activity necessary for the formation of peripheral neurons in Drosophila. Genes & Dev 843–852
Caudy M, Vaessin H, Brand M, Tuma R, Jan LY, Jan YN (1988b) Daughterless, a gene essential for both neurogenesis and sex determination in Drosophila, has sequence similarities to myc and the achaete-scute complex. Cell 55:1061–1067
Cavener DR (1987) Comparison of the consensus sequence flanking translation start sites in Drosophila and vertebrates. Nucleic Acids Res 15:1353–1361
Church GM, Gilbert W (1984) Genomic sequencing. PNAS 81:1991–1995
Ellis HM, Spann DR, Posakony JW (1990) Extramacrochaetae, a negative regulator of sensory organ development in Drosophila, defines a new class of helix-loopp-helix proteins. Cell 61:27–38
Ferreiro B, Skoglund P, Bailey A, Dorsky R, Harris WA (1994) XASH1, a Xenopus homolog of achaete-scute: a proneural gene in anterior regions of the vertebrate CNS. Mech Dev 40:25–36
Franco del Amo F, Gendron-Maguire M, Swiatek PJ, Gridley T (1993) Cloning, sequencing and expression of the mouse mammalian achaete-scute homolog 1 (MASH1). Biochem Biophys Acta 1171:323–327
Garrell J, Modolell J (1990) The Drosophila extramacrochaetae locus, an antagonist of proneural genes that, like these genes, encodes a helix-loop-helix protein. Cell 61:39–48
Gonzalez F, Romani S, Cubas C, Modolell J, Campuzano S (1989) Molecular analysis of the asense gene, a member of the achaete-scute complex of Drosophila melanogaster, and its novel role in optic lobe development. EMBO J 8:3409–3417
Ghysen A, Dambly-Chaudière C (1988) From DNA to form: the achaete-scute complex. Genes Dev 2:495–501
Guillemot F, Lo L-C, Johnson JE, Auerbach A, Anderson DJ, Joyner AL (1993) Mammalian achaete-scute homolog 1 is required for the early development of olfactory and autonomic neurons. Cell 75:463–476
Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–67
Jasoni CL, Walker MB, Morris MD, Reh TA (1994) A chicken achaete-scute homolog (CASH1) is expressed in a temporally and spatially discrete manner in the developing nervous system. Development 120:769–783
Jiménez F, Campos-Ortega JA (1979) On a region of the Drosophila genome necessary for central nervous system development. Nature 282:310–312
Jiménez F, Campos-Ortega JA (1987) Genes in subdivision 1B of the Drosophila melanogaster X-chromosome and their influence on neural development. J Neurogenet 4:179–200
Johnson JE, Birren SJ, Anderson DJ (1990) Two rat homologues of Drosophila achaete-scute specifically expressed in neuronal precursors. Nature 346:858–867
Kirby ML (1988) Nodose placode contributes autonomic neurons to the heart in the absence of cardiac neural crest. J Neurosci 8:1089–1095
Kirby ML (1993) Cellular and molecular contributions of the cardiac neural crest to cardiovascular development. Trends Cardiovasc Med 3:18–23
Klämbt C, Knust E, Tietze K, Campos-Ortega JA (1989) Closely related transcripts encoded by the nurogenic gene complex Enhancer of split of Drosophila melanogaster. EMBO J 8:203–211
Lo L-C, Johnson JE, Wuenschell CW, Saito T, Anderson DJ (1991) Mammalian achaete-scute homolog 1 is transiently expressed by spatially restricted subsets of early neuroepithelial and neural crest cells. Genes Dev 5:1524–1537
Murre C, McCaw PS, Vaessin H, Candy M, Jan LY, Jan YN, Cabrera CV, Buskin JN, Hauschka SD, Lassar AB, Weintraub H, Baltimore D (1980) Interactions between heterologous helixloop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell 58:537–544
Murre C, McCaw PS, Baltimore D (1989) A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins. Cell 56:777–783
Rosen B, Beddington RSP (1993) Whole-mount in situ hybridization in the mouse embryo: gene expression in three dimensions. TIG 9:162–167
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. PNAS 74:5463–5467
Tapscott SJ, Davis RL, Thayer MJ, Chang PF, Weintraub H, Lassar AB (1988) MyoD: a nuclear phosphoprotein requiring a Myc homology region to convert fibroblasts to myoblasts. Science 242:405–411
Thisse B, Stoetzel C, Gorostiza-Thisse C, Perrin-Schmitt F (1988) Sequence of the twist gene and nuclear location of its protein in endomesodermal cells of early Drosophila embryos. EMBO J 7:2175–2183
Villares R, Cabrera CV (1987) The achaete-scute gene complex of D. melanogaster: conserved domains in a subset of genes required for neurogenesis and their homology to myc. Cell 50:415–424
Wilkinson DG (1992) Rickwood D, Hames BD (eds) In situ hybridization: a practical approach. IRS Press, Oxford, pp 75–83
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wang, S., Kirby, M.L. Expression pattern of chick achaete-scute homologue (CASH) associated with mid-embryogenesis (stages 23 to 28). Roux's Arch Dev Biol 205, 1–9 (1995). https://doi.org/10.1007/BF00188838
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00188838