Abstract
The individual cells of the Xenopus cleavage-stage embryo have been fate mapped, revealing which of these cells contribute to the retina. Using this retina fate map, one can specifically modulate levels of gene expression in retina lineages to determine the function of proteins in various aspects of early retinal development, such as formation of the eye fields and determination of specific cell fates. This chapter presents the techniques for identifying specific retina blastomere precursor cells, and injecting them with lineage tracers, mRNAs encoding wild-type and mutant constructs or morpholino antisense oligonucleotides to alter gene expression.
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References
Dale L, Slack JMW (1987) Fate map of the 32-cell stage of Xenopus laevis. Development 100:279–295
Moody SA (1987) Fates of the blastomeres of the 16-cell stage Xenopus embryo. Dev Biol 119:560–578
Moody SA (1987) Fates of the blastomeres of the 32-cell stage Xenopus embryo. Dev Biol 122:300–319
Moody SA, Kline MJ (1990) Segregation of fate during cleavage of frog (Xenopus laevis) blastomeres. Anat Embryol 182:347–362
Moody SA (1989) Quantitative lineage analysis of the origin of frog primary motor and sensory neurons from cleavage stage blastomeres. J Neurosci 9:2919–2930
Huang S, Moody SA (1992) Does lineage determine the dopamine phenotype in the tadpole hypothalamus: a quantitative analysis. J Neurosci 12:1351–1362
Huang S, Moody SA (1993) The retinal fate of Xenopus cleavage stage progenitors is dependent upon blastomere position and competence: studies of normal and regulated clones. J Neurosci 13:3193–3210
Kenyon KL, Zaghloul N, Moody SA (2001) Transcription factors of the anterior neural plate alter cell movements of epidermal progenitors to specify a retinal fate. Dev Biol 240:77–91
Huang S, Moody SA (1995) Asymmetrical blastomere origin and spatial domains of dopamine and Neuropeptide Y amacrine cells in Xenopus tadpole retina. J Comp Neurol 360:2–13
Huang S, Moody SA (1997) Three types of serotonin-containing amacrine cells in the tadpole retina have distinct clonal origins. J Comp Neurol 387:42–52
Moore KB, Moody SA (1999) Animal-vegetal asymmetries influence the earliest steps in retinal fate commitment in Xenopus. Dev Biol 212:25–41
Guthrie S, Turin L, Warner AE (1988) Patterns of junctional communication during development of the early amphibian embryo. Development 103:769–783
Weisblat DA, Sawyer RT, Stent GS (1978) Cell lineage analysis by intracellular injection of a tracer enzyme. Science 202:1295–1298
Jacobson M (1985) Clonal analysis and cell lineages of the vertebrate nervous system. Annu Rev Neurosci 8:71–102
Stent GS, Weisblat DA (1985) Cell lineage in the development of invertebrate nervous systems. Annu Rev Neurosci 8:45–70
Gimlich RL, Braun J (1985) Improved fluorescent compounds for tracing cell lineage. Dev Biol 109:509–514
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805
Sive HL, Grainger RM, Harland RM (2000) Early development of Xenopus laevis. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Vincent J-P, Gerhart JC (1987) Subcortical rotation in Xenopus eggs: an early step in embryonic axis specification. Dev Biol 123:526–539
Klein SL (1987) The first cleavage furrow demarcates the dorsal–ventral axis in Xenopus embryos. Dev Biol 120:299–304
Masho R (1990) Close correlation between the first cleavage plane and the body axis in early Xenopus embryos. Dev Growth Differ 32:57–64
Hainski AM, Moody SA (1992) Xenopus maternal RNAs from a dorsal animal blastomere induce a secondary axis in host embryos. Development 116:347–355
Peng HB (1991) Appendix A: solutions and protocols. Methods Cell Biol 36:657–662
Nakamura O, Kishiyama K (1971) Prospective fates of blastomeres at the 32-cell stage of Xenopus laevis embryos. Proc Jpn Acad 47:407–412
Hirose G, Jacobson M (1979) Clonal organization of the central nervous system of the frog. I. Clones stemming from individual blastomeres of the 16-cell and earlier stages. Dev Biol 71:191–202
Jacobson M, Hirose G (1981) Clonal organization of the central nervous system of the frog. II. Clones stemming from individual blastomeres of the 32- and 64-cell stages. J Neurosci 1:271–284
Sullivan SA, Moore KB, Moody SA (1999) Early events in blastomere fate determination. In: Moody SA (ed) Cell lineage and cell fate determination. Academic, New York, pp 297–321
Nieuwkoop PD, Faber J (1967) Normal table of Xenopus laevis (Daudin). Elsevier-North Holland Publishing Co., Amsterdam
Acknowledgement
This work was supported by NSF grant IOS-0817902.
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Moody, S.A. (2012). Targeted Microinjection of Synthetic mRNAs to Alter Retina Gene Expression in Xenopus Embryos. In: Wang, SZ. (eds) Retinal Development. Methods in Molecular Biology, vol 884. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-848-1_6
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DOI: https://doi.org/10.1007/978-1-61779-848-1_6
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