Molecular cloning and expression analysis of human retinal NeuroD mRNA

H. R. Acharya, Iqbal Ahmad

Research output: Contribution to journalArticle

Abstract

PURPOSE: The activation of photoreceptor specific gene is an essential step towards differentiation and maturation of photoreceptors. This step is likely to be regulated by a combinatorial effect of different transcription factors. Alteration or absence of the expression of these transcription factors can compromise the maintenance of the differentiated properties and eventually lead to degenerative changes. We have recently shown th.it, NeuroD a bHLH transcription factor may participate in the terminal differential on of retinal neurons in the rat (Ahmad, Soc. Neurosci. Abst. 22:1978, 1996). To investigate the role of NeuroD in human retinal development and degenerative changes we have begun the identification and analysis of NeuroD mRNA in adult human rttina. METHODS: cDNA corresponding to human NeuroD mRNA was characterized by homology cloning using degenerate primers and adult human retinal cDNA library. Northern and in situ hybridization was carried out using NeuroD cDNA as a probe. Western and immunocytochemical analyses were carried out using antibody raised against mouse NeuroD peptide. In addition, NeuroD expression was inalyzed in fetal monkey retina (1I7GD). RESULTS: The cDNA (GenBank accession #118057) cloned from adult human retina was homologous to mouse (Lee.et.al.,Science 268:836 1995) and rat (GenBank accession #U80603) NeuioD cDNA. NeuroD transcripts (~3kb) were detected in adult human retina and the) were primarily distributed in the outer nuclear layer. In both adult human and fetal rionkey retina NeuroD immunoreactivity was localized in the outer nuclear layer. CONCLUSION: The expression analyses of NeuroD suggest that it may play role in retinal development and in the maintenance of differentiated properties of retinal cell types, particularly that of photoreceptors. As such alterations in the expression of NeuroD may play a role in the degenerative process such as those observed in rethitis piementosa.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume38
Issue number4
StatePublished - Dec 1 1997

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Molecular Cloning
Messenger RNA
Retina
Complementary DNA
Nucleic Acid Databases
Transcription Factors
Maintenance
Basic Helix-Loop-Helix Transcription Factors
Retinal Neurons
Vertebrate Photoreceptor Cells
Essential Genes
Human Development
Gene Library
Haplorhini
In Situ Hybridization
Organism Cloning
Peptides
Antibodies

ASJC Scopus subject areas

  • Ophthalmology

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Molecular cloning and expression analysis of human retinal NeuroD mRNA. / Acharya, H. R.; Ahmad, Iqbal.

In: Investigative Ophthalmology and Visual Science, Vol. 38, No. 4, 01.12.1997.

Research output: Contribution to journalArticle

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