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Analytical Post-Embedding Immunogold–Electron Microscopy with Direct Gold-Labelled Monoclonal Primary Antibodies against RIBEYE A- and B-Domain Suggests a Refined Model of Synaptic Ribbon Assembly
by
Stella Papadopoulos
Stella Papadopoulos 1,
René Tinschert
René Tinschert 1,
Iason Papadopoulos
Iason Papadopoulos 2,
Xenia Gerloff
Xenia Gerloff 2 and
Frank Schmitz
Frank Schmitz 1,*
1
Institute of Anatomy, Department of Neuroanatomy, Medical School, Saarland University, 66421 Homburg, Germany
2
Mathematical Institute, University of Bonn, 53115 Bonn, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2024, 25(13), 7443; https://doi.org/10.3390/ijms25137443 (registering DOI)
Submission received: 29 May 2024
/
Revised: 2 July 2024
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Accepted: 4 July 2024
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Published: 6 July 2024
Abstract
Synaptic ribbons are the eponymous specializations of continuously active ribbon synapses. They are primarily composed of the RIBEYE protein that consists of a unique amino-terminal A-domain and carboxy-terminal B-domain that is largely identical to the ubiquitously expressed transcriptional regulator protein CtBP2. Both RIBEYE A-domain and RIBEYE B-domain are essential for the assembly of the synaptic ribbon, as shown by previous analyses of RIBEYE knockout and knockin mice and related investigations. How exactly the synaptic ribbon is assembled from RIBEYE subunits is not yet clear. To achieve further insights into the architecture of the synaptic ribbon, we performed analytical post-embedding immunogold–electron microscopy with direct gold-labelled primary antibodies against RIBEYE A-domain and RIBEYE B-domain for improved ultrastructural resolution. With direct gold-labelled monoclonal antibodies against RIBEYE A-domain and RIBEYE B-domain, we found that both domains show a very similar localization within the synaptic ribbon of mouse photoreceptor synapses, with no obvious differential gradient between the centre and surface of the synaptic ribbon. These data favour a model of the architecture of the synaptic ribbon in which the RIBEYE A-domain and RIBEYE B-domain are located similar distances from the midline of the synaptic ribbon.
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MDPI and ACS Style
Papadopoulos, S.; Tinschert, R.; Papadopoulos, I.; Gerloff, X.; Schmitz, F.
Analytical Post-Embedding Immunogold–Electron Microscopy with Direct Gold-Labelled Monoclonal Primary Antibodies against RIBEYE A- and B-Domain Suggests a Refined Model of Synaptic Ribbon Assembly. Int. J. Mol. Sci. 2024, 25, 7443.
https://doi.org/10.3390/ijms25137443
AMA Style
Papadopoulos S, Tinschert R, Papadopoulos I, Gerloff X, Schmitz F.
Analytical Post-Embedding Immunogold–Electron Microscopy with Direct Gold-Labelled Monoclonal Primary Antibodies against RIBEYE A- and B-Domain Suggests a Refined Model of Synaptic Ribbon Assembly. International Journal of Molecular Sciences. 2024; 25(13):7443.
https://doi.org/10.3390/ijms25137443
Chicago/Turabian Style
Papadopoulos, Stella, René Tinschert, Iason Papadopoulos, Xenia Gerloff, and Frank Schmitz.
2024. "Analytical Post-Embedding Immunogold–Electron Microscopy with Direct Gold-Labelled Monoclonal Primary Antibodies against RIBEYE A- and B-Domain Suggests a Refined Model of Synaptic Ribbon Assembly" International Journal of Molecular Sciences 25, no. 13: 7443.
https://doi.org/10.3390/ijms25137443
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