June 1997
Volume 38, Issue 7
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Articles  |   June 1997
Pax-6, Prox 1, and Chx10 homeobox gene expression correlates with phenotypic fate of retinal precursor cells.
T Belecky-Adams; S Tomarev; H S Li; L Ploder; R R McInnes; O Sundin; R Adler
Author Affiliations
  • T Belecky-Adams
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • S Tomarev
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • H S Li
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • L Ploder
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • R R McInnes
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • O Sundin
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
  • R Adler
    Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287-9257, USA.
Investigative Ophthalmology & Visual Science June 1997, Vol.38, 1293-1303. doi:
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      T Belecky-Adams, S Tomarev, H S Li, L Ploder, R R McInnes, O Sundin, R Adler; Pax-6, Prox 1, and Chx10 homeobox gene expression correlates with phenotypic fate of retinal precursor cells.. Invest. Ophthalmol. Vis. Sci. 1997;38(7):1293-1303.

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Abstract

PURPOSE: To study the expression patterns of the homeobox genes Pax-6, Prox 1, and Chx10 during chick retinal development in vivo and in vitro. METHODS: Sections of paraformaldehyde-fixed, paraffin-embedded eyes were obtained at a range of developmental stages. In situ hybridization was carried out on tissue sections using digoxigenin-labeled sense and antisense RNA probes that recognize chicken Pax-6 and Prox 1 (whose sequences were already available), and chicken Chx10 (which was cloned and sequenced as part of this study). Selected developmental stages were also studied by immunocytochemistry with antibodies against Pax-6 and Prox 1, and by Northern blot analysis using 32P-labeled probes. RESULTS: Until embryonic day (ED) 5, in situ hybridization shows widespread, diffuse distribution of all three genes. Between ED 6 and ED 8, however, they acquire distinct, topographically specific patterns of expression. The Prox 1 signal is predominantly expressed in the prospective horizontal cell layer of the neuroepithelium, decreases vitreally, and is absent from ganglion cells and the prospective photoreceptor layer. Pax-6 is strongly expressed only in the prospective ganglion-cell and amacrine-cell regions at the same stages, and is not detected in prospective photoreceptors. Chx10 expression becomes concentrated in the future bipolar-cell region of the inner nuclear layer. Similar patterns are maintained by ED 15 through ED 18, after cell differentiation has taken place. Pax-6 and Prox 1 immunoreactive materials showed nuclear localization and a pattern of laminar distribution equivalent to that seen by in situ hybridization. CONCLUSIONS: These results suggest that the differentiated fate of retinal precursor cells may be influenced by Pax-6, Prox 1, or Chx10, this hypothesis is now being tested using dissociated chick embryo retinal cell cultures.

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