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  • Ascidian embryos  (1)
  • Muscle-specific expression  (1)
  • UV-sensitivity  (1)
  • 1995-1999  (2)
  • 1
    ISSN: 1432-041X
    Keywords: Ascidians ; Egg fragments ; Muscle determinants ; UV-sensitivity ; Maternal messenger RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ascidian egg contains cytoplasmic determinants that specify the fate of larval muscle cells. In a previous study, we developed an experimental system to identify the molecular nature of muscle determinants, in which unfertilized Ciona savignyi eggs were fragmented into four pieces by centrifugation. When inseminated, only nucleated fragments (red fragments) develop into partial embryos that only show differentiation of epidermal cells. One type of enucleated fragment (black fragment) has the remarkable ability to promote muscle differentiation when fused with red fragments. In the present study, using this experimental system, we investigated the molecular nature of muscle determinants. UV irradiation of black fragments suppressed the ability to promote expression of the muscle-specific protein, myosin heavy chain. The wavelength of UV light responsible for the inactivation (250–275 nm) suggested that UV-sensitive targets are nucleic acids. Injection of poly(A)+ RNA isolated from an un-irradiated black-fragment-rich fraction into UV-irradiated black fragments partially recovered the ability to promote the expression of myosin heavy chain protein. Poly(A)+ RNA from a red-fragment-rich fraction did not rescue the suppression of UV-irradiated black fragments. These results suggest that maternal mRNAs enriched in black fragments are closely associated with muscle determinants in the ascidian egg.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Development genes and evolution 206 (1996), S. 54-63 
    ISSN: 1432-041X
    Keywords: Key words cis-Elements ; Myosin heavy-chain gene ; Muscle-specific expression ; Ascidian embryos
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The B-line muscle cells of the ascidian embryo are specified autonomously depending on determinants prelocalized in the myoplasm of unfertilized eggs. Expression of muscle-specific actin and myosin heavy-chain genes commences in the B-line presumptive muscle cells as early as the 32-cell stage. To explore the intrinsic genetic program for this differentiation, we analysed cis-regulatory elements of the Halocynthia roretzi muscle myosin heavy-chain gene (HrMHC1). Comparison of the entire amino acid sequence of HrMHC1 with those of other invertebrates and vertebrates indicated that HrMHC1 resembles myosin heavy-chain of vertebrate skeletal and cardiac muscles. A fusion gene was constructed consisting of 132 bp upstream the 5′-end of HrMHC1 gene fused to a bacterial lacZ reporter. When the fusion gene was microinjected into fertilized eggs, the reporter gene was eventually expressed only in muscle cells of tailbud embryos. It has been reported that 103 bp of sequence 5′ of the transcription start site of the ascidian embryonic muscle actin gene (HrMA4) contains information sufficient for muscle-specific expression (Hikosaka et al. 1994). Comparison of the 132 bp of sequence 5′ of the HrMHC1 gene with the 103 bp of sequence 5′ of the HrMA4 gene revealed several common motifs shared by the two genes (E-box, GATA box and Boxes A, B, T1 and T2). Point mutations inserted into these motifs suggested that the Box T1/T2 (TTTTTTCTTTCA) is critical for the promoter activity of the HrMHC1 gene.
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