The transcriptome of retinal Müller glial cells.
Bottom Line: Genes expressed exclusively by Müller glia were identified as novel markers.In addition, a novel BAC transgenic mouse that expresses Cre in Müller glia cells was generated.The molecular fingerprint of Müller glia provides a foundation for further studies of Müller glia development and function in normal and diseased states.
Affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.Show MeSH
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Mentions: Adult murine retinae were dissociated to single cells by using papain digestion. Within less than 2 hours of dissection, individual Müller glial cells were identified by their distinctive morphology (Fig. 1) and picked from a dish of dissociated cells with a micropipette. They were then washed, lysed, and subjected to reverse transcription. After a terminal transferase reaction to add A's to the 3′ end of the cDNA, 35 rounds of PCR were carried out by using modified oligo dT primers. cDNA preparations from five individual Müller glial cells were hybridized to Affymetrix Genechip® Mouse genome 430 2.0 arrays. These arrays provide almost complete coverage of the mouse genome with 45,000 probe sets. The identity of cells as Müller glial cells was confirmed by expression of known marker genes for Müller glia. Glutamine synthetase (Glul), clusterin (Clu), dickkopf homolog 3 (Dkk3) (Blackshaw et al., 2004), and S100 calcium binding protein A16 (Seigel et al., 1996) were successfully detected (Fig. 2 and Supplemental Fig. S1). All of the cells share expression of these key marker transcripts.
Affiliation: Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.