Combinatorial regulation of a Blimp1 (Prdm1) enhancer in the mouse retina

• Published in: PloS one 2017-08, Vol.12 (8), p.e0176905
• Main Authors: Mills, Taylor S, Eliseeva, Tatiana, Bersie, Stephanie M, Randazzo, Grace, Nahreini, Jhenya, Park, Ko Uoon, Brzezinski, 4th, Joseph A
• Format: Article
• Language: English
• Subjects: Animals; Binding sites; Biology and Life Sciences; Bipolar cells; Cell cycle; Cloning; Combinatorial analysis; Deoxyribonuclease; Enhancer Elements, Genetic; Gene expression; Genetic regulation; Genomes; Health aspects; Hypersensitivity; Interneurons; Medicine and Health Sciences; Mice; Mice, Inbred C57BL; Mutagenesis; Neurogenesis; Otx2 protein; Photoreception; Photoreceptors; Plasmids; Positive Regulatory Domain I-Binding Factor 1; Research and Analysis Methods; Retina; Retina - metabolism; Retinoic acid; Sequences; Social Sciences; Stochasticity; Temporal variations; Transcription (Genetics); Transcription factors; Transcription Factors - genetics; Transgenic mice; Zinc finger proteins
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0176905

The mouse retina comprises seven major cell types that exist in differing proportions. They are generated from multipotent progenitors in a stochastic manner, such that the relative frequency of any given type generated changes over time. The mechanisms determining the proportions of each cell type are only partially understood. Photoreceptors and bipolar interneurons are derived from cells that express Otx2. Within this population, Blimp1 (Prdm1) helps set the balance between photoreceptors and bipolar cells by suppressing bipolar identity in most of the cells. How only a subset of these Otx2+ cells decides to upregulate Blimp1 and adopt photoreceptor fate is unknown. To understand this, we investigated how Blimp1 transcription is regulated. We identified several potential Blimp1 retinal enhancer elements using DNase hypersensitivity sequencing. Only one of the elements recapitulated Blimp1 spatial and temporal expression in cultured explant assays and within the retinas of transgenic mice. Mutagenesis of this retinal Blimp1 enhancer element revealed four discrete sequences that were each required for its activity. These included highly conserved Otx2 and ROR (retinoic acid receptor related orphan receptor) binding sites. The other required sequences do not appear to be controlled by Otx2 or ROR factors, increasing the complexity of the Blimp1 gene regulatory network. Our results show that the intersection of three or more transcription factors is required to correctly regulate the spatial and temporal features of Blimp1 enhancer expression. This explains how Blimp1 expression can diverge from Otx2 and set the balance between photoreceptor and bipolar fates.