Biology subjects

Noel, N. C. L.

Publications and source records attributed to Noel, N. C. L..

2 recordsLinked to original sources

bioRxiv2025

Polysialic acid is a versatile marker for retinal Müller glia in common vertebrate model organisms and systems

Muller glia are retinal support cells that play crucial roles in tissue structure, waste management, and repair. A challenge for the field has been to find Muller glia markers that detect non-reactive cells or work well in non-mammalian models. We introduce two novel markers for identifying reactive and non-reactive Muller glia in vertebrate retinas: a modified enzyme lectin (GFP-EndoNDM) and a monoclonal antibody (mAb735). These markers recognize polysialic acid (polySia), which is a highly conserved glycosylation modification in humans and vertebrates. In the retina, polySia is present on Muller glia predominantly in the form of polySia-NCAM. We used GFP-EndoNDM and mAb735 to investigate polySia distribution in Muller glia of fish, amphibians, reptiles, birds, rodents, retinal organoids, and humans. In adult retinas of most species, polySia was localized to the Muller glia and spanned outer to inner retina, with inner plexiform layer (IPL) sublaminae ramifications. Gliosis was also detectable in degenerating murine retinas. Notable species differences were that only outer retinal regions of Muller glia were labelled in adult zebrafish, whereas the outer Muller glia body up to the first IPL sublamina was labelled in adult turquoise killifish. There was no significant retinal polySia labeling in larval zebrafish, but it was present in the brain. Larval turquoise killifish have polySia throughout the retina, similar to other adult vertebrates. Labelling polySia expands the scientific toolbox for Muller glia markers, and offers a versatile way to visualize and monitor structural changes in non-reactive and reactive Muller glia across most vertebrate species. HighlightsO_LIPolySia is a highly conserved Muller glia marker in vertebrate retina and retinal organoids C_LIO_LILabeling Muller glia with markers of polySia, facilitates the analysis of fine Muller glia processes in the IPL sublaminae, whole Muller glia morphology, and degenerative gliosis C_LIO_LIPolySia can be labeled with an engineered lectin conjugate (GFP-EndoNDM) and a commercially-available monoclonal antibody C_LI

cell biology↗

bioRxiv2024

Iterative Bleaching Extends Multiplexity (IBEX) imaging facilitates simultaneous identification of all cell types in the vertebrate retina

The vertebrate retina is a complex multicellular tissue made up of distinct neuron types and glia, arranged in a stereotypic layered organisation to facilitate vision. Understanding how these cell types come together to form precise circuits during development requires the ability to simultaneously discriminate between multiple cell types and their spatial position in the same tissue. Currently, we have a limited capacity to resolve all constitutive cell types and their relationships to one another due to our limited ability to combine multiple cellular markers. To extend this capacity, we have adapted a highly multiplexed immunohistochemistry technique known as Iterative Bleaching Extends Multiplexity (IBEX) and applied it to the development of the zebrafish (Danio rerio) retina. IBEX allows for multiple rounds of cellular labelling to be performed, before imaging and integration of data, resulting in the ability to visualise multiple markers on the same tissue. We have optimised IBEX in zebrafish using fluorescent micro-conjugation of known antibody markers to label the complete retina with up to 11 cell-specific antibodies. We have further adapted the IBEX technique to be compatible with fluorescent transgenic reporter lines, in situ hybridisation chain reaction (HCR), and wholemount immunohistochemistry (WMIHC). We then took advantage of IBEX to explore the multicellular relationships in the developing retina between glial cells and neurons and photoreceptor subtypes. Finally, we tested IBEX on retinas from the emerging ageing model, the killifish (Nothobranchius furzeri), and developmental model, the African clawed frog (Xenopus laevis), demonstrating the usefulness of the technique across multiple species. The techniques described here can be applied to any tissue in any organism where antibodies are readily available to efficiently explore cellular relationships in the context of development, ageing or disease.

developmental biology↗