Ankyrins are essential for synaptic integrity of photoreceptors in the mouse outer retina

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ABSTRACT Retinal circuit assembly relies on the precise timing and positioning of key molecules between neuronal partners to mediate proper synapse formation. In the outer retina, horizontal cells are important interneurons that make the first contacts to photoreceptors and begin to segregate visual information into two distinct pathways by selectively forming synapses to the different types of photoreceptors. Dendrites of horizontal cells synapse exclusively to cone photoreceptors whereas the axon terminal synapses to rod photoreceptors. Failure to properly form these early connections disrupts the downstream connectivity of other postsynaptic neurons and leads to abnormal visual function. Although these early events are critical for proper synapse development, little is known about the molecular mechanisms that establish horizontal cell to photoreceptor connectivity during development. In the present study, we performed single-cell RNA sequencing and uncovered new molecules that are highly expressed in horizontal cells. These include different members of the cytoskeletal scaffolding family of Ankyrins that are known to form specialized regions within neurons by recruiting different molecules to the membrane and linking them to the cytoskeleton. Specifically, we found Ankyrin-B to be highly expressed in horizontal cells at early time points and Ankyrin-G to be expressed at later stages. Loss of both Ankyrin-B and Ankyrin-G leads to synaptic defects between horizontal cells and photoreceptors and disrupts in vivo retinal responses. In summary, our findings uncovered a new role for Ankyrins in mediating synaptic connectivity between horizontal cells and photoreceptors required for normal visual function. SIGNIFICANCE STATEMENT In the mammalian retina, the first synapse between photoreceptors and their downstream targets begins to separate visual information into two distinct pathways. During retinal development, photoreceptors first make contacts to horizontal cells in a temporal- and spatial-specific manner. Although this initial contact is critical for synaptogenesis, little is known about the key molecules responsible for selective wiring of horizontal cells to photoreceptors. In this study, we performed single-cell RNA sequencing and identified the family of cytoskeletal scaffolding proteins Ankyrins to be differentially expressed in horizontal cells. Loss of Ankyrins impairs synaptic connectivity between horizontal cells and their photoreceptor partners leading to abnormal visual responses. Taken together, our work uncovered a new function of Ankyrins at photoreceptor synapses.
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Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results Ankyrins are essential at the photoreceptor synapse in the mouse outer retina Elizabeth Zuniga-Sanchez , Ross M Perez , Jay Campbell , Martina Cavallini , Collin Todora , Daniela Becerril , Debalina Goswami-Sewell , Rajashree Venkatraman , Cesiah C Gomez , Caitlin Bagnetto , Audrey Lee , Marlon F Mattos , Yi-Rong Peng , Mrinalini Hoon doi: https://doi.org/10.1101/2025.02.11.637690 Elizabeth Zuniga-Sanchez 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site For correspondence: elizabeth.zuniga-sanchez{at}bcm.edu Ross M Perez 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Jay Campbell 2 University of Wisconsin-Madison; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Martina Cavallini 3 UCLA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Collin Todora 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Daniela Becerril 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Debalina Goswami-Sewell 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Rajashree Venkatraman 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Cesiah C Gomez 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Caitlin Bagnetto 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Audrey Lee 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Marlon F Mattos 1 Baylor College of Medicine; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Yi-Rong Peng 3 UCLA Find this author on Google Scholar Find this author on PubMed Search for this author on this site Mrinalini Hoon 2 University of Wisconsin-Madison; Find this author on Google Scholar Find this author on PubMed Search for this author on this site Abstract Info/History Metrics Supplementary material Preview PDF Abstract Retinal circuit assembly relies on the precise timing and positioning of key molecules between neuronal partners to mediate proper synapse formation. In the outer retina, horizontal cells are important interneurons that make the first contacts to photoreceptors and begin to segregate visual information into two distinct pathways by selectively forming synapses to the different types of photoreceptors. Dendrites of horizontal cells synapse exclusively to cone photoreceptors whereas the axon terminal synapses to rod photoreceptors. Failure to properly form these early connections disrupts the downstream connectivity of other postsynaptic neurons and leads to abnormal visual function. Although these early events are critical for proper synapse development, little is known about the molecular mechanisms that establish horizontal cell to photoreceptor connectivity during development. In the present study, we performed single-cell RNA sequencing and uncovered new molecules that are highly expressed in horizontal cells. These include different members of the cytoskeletal scaffolding family of Ankyrins that are known to form specialized regions within neurons by recruiting different molecules to the membrane and linking them to the cytoskeleton. Specifically, we found Ankyrin-B to be highly expressed in horizontal cells at early time points and Ankyrin-G to be expressed at later stages. Loss of both Ankyrin-B and Ankyrin-G leads to synaptic defects between horizontal cells and photoreceptors and disrupts in vivo retinal responses. In summary, our findings uncovered a new role for Ankyrins in mediating synaptic connectivity between horizontal cells and photoreceptors required for normal visual function. Competing Interest Statement The authors have declared no competing interest. Footnotes New developmental analysis of horizontal cell specificity to photoreceptors in Figure 1 and single-cell RNA sequencing data in Figure 2. Funder Information Declared National Eye Institute , F31EY035931 , R01EY033037 , R01EY035324 , T32EY007026 , P30EY002520 , P30EY00331 Research to Prevent Blindness Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission. View the discussion thread. Back to top Previous Next Posted May 23, 2026. Download PDF Supplementary Material Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. 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