The Calcineurin-Binding, Activity-Dependent Splice Variant Dynamin1xb Is Highly Enriched in Synapses in Various Regions of the Central Nervous System

Eich, Marie‐Lisa; Dembla, Ekta; Wahl, Silke; Dembla, Mayur; Schwarz, Karin; Schmitz, Frank

doi:10.3389/fnmol.2017.00230

Cited by 7 publications

(24 citation statements)

References 136 publications

(280 reference statements)

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“…Additional primary antibodies used in the study: anti‐visual arrestin [Santa Cruz (E12, sc‐34547)] affinity‐purified goat polyclonal antibody against a peptide from an internal region of human visual arrestin, used in a 1:250 dilution for immunofluorescence microscopy; anti‐mGluR6; rabbit polyclonal antiserum (Katiyar et al , ) used in a 1:200 dilution for immunofluorescence microscopy; anti‐Cask mouse monoclonal antibody (Anjum et al , ), used in a 1:200 dilution for immunofluorescence microscopy; anti‐RIM1/2 (Anjum et al , ); rabbit polyclonal antiserum used in a 1:500 dilution for immunofluorescence microscopy; anti‐dynamin1xb (clone 9E10; Eich et al , ), mouse monoclonal antibody used at a 1:300 dilution for Western blot and immunofluorescence microscopy; anti‐PSD95 (NeuroMABs) used in a 1:200 dilution for immunofluorescence microscopy; anti‐synaptophysin (SIGMA, clone SVP‐38; #SAB4200544) mouse monoclonal antibody against synaptophysin, purified immunoglobulin, used at a 1:1,000 dilution for Western blots; anti‐mCherry, mouse monoclonal antibody (Abcam 1C51, ab125096) used at 1:2,000 dilution for Western blot analyses; Anti‐Na v ‐channel pan mouse monoclonal antibody (Sigma, clone K58/35; S8809) used at dilution of 1:50 for immunolabeling.…”

Section: Methodsmentioning

confidence: 99%

“…Triple‐immunolabeling analyses with three different primary antibodies (with two antibodies from an identical species; i.e., mouse or rabbit) were performed, as previously described (Eich et al , ). In brief, two of the three primary antibodies that were generated in different species (i.e., a mouse primary antibody and a rabbit primary antibody) were incubated simultaneously overnight at 4°C at the indicated dilutions.…”

Section: Methodsmentioning

confidence: 99%

“…SDS–PAGE and Western blotting experiments were performed as previously described (Schmitz et al , ; Wahl et al , ; Eich et al , ). Protein determination of retinal samples dissolved in Laemmli SDS sample buffer: Mouse retinas were isolated 5 min post‐mortem and dissolved in 100 μl hot (96°C) Laemmli sample buffer.…”

Section: Methodsmentioning

confidence: 99%

“…Triple-immunolabeling analyses with three different primary antibodies (with two antibodies from an identical species; i.e., mouse or rabbit) were performed, as previously described (Eich et al, 2017).…”

Section: Triple Immunolabeling Of Semi-thin Sectionsmentioning

confidence: 99%

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Early auto‐immune targeting of photoreceptor ribbon synapses in mouse models of multiple sclerosis

et al. 2018

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Optic neuritis is one of the first manifestations of multiple sclerosis. Its pathogenesis is incompletely understood, but considered to be initiated by an auto‐immune response directed against myelin sheaths of the optic nerve. Here, we demonstrate in two frequently used and well‐validated mouse models of optic neuritis that ribbon synapses in the myelin‐free retina are targeted by an auto‐reactive immune system even before alterations in the optic nerve have developed. The auto‐immune response is directed against two adhesion proteins (CASPR1/CNTN1) that are present both in the paranodal region of myelinated nerves as well as at retinal ribbon synapses. This occurs in parallel with altered synaptic vesicle cycling in retinal ribbon synapses and altered visual behavior before the onset of optic nerve demyelination. These findings indicate that early synaptic dysfunctions in the retina contribute to the pathology of optic neuritis in multiple sclerosis.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Triple Immunolabeling Of Semi-thin Sectionsmentioning

confidence: 99%

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Early auto‐immune targeting of photoreceptor ribbon synapses in mouse models of multiple sclerosis

et al. 2018

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“…Immunolabeling of resin sections. For confocal imaging, immunolabeling was performed using 0.5 μm thin ("semi-thin") resin sections that were placed on glass coverslips, as previously described 36,37,44,47 . For Super-Resolution Structured Illumination Microscopy (SR-SIM) imaging, 1.5 μm thin resin sections were used to cover the entire synaptic terminal of a rod synapse.…”

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confidence: 99%

Synaptic ribbons foster active zone stability and illumination-dependent active zone enrichment of RIM2 and Cav1.4 in photoreceptor synapses

Dembla

Maxeiner

et al. 2020

Sci Rep

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Rod photoreceptor synapses use large, ribbon-type active zones for continuous synaptic transmission during light and dark. Since ribbons are physically connected to the active zones, we asked whether illumination-dependent changes of ribbons influence Cav1.4/RIM2 protein clusters at the active zone and whether these illumination-dependent effects at the active zone require the presence of the synaptic ribbon. We found that synaptic ribbon length and the length of presynaptic Cav1.4/RIM2 clusters are tightly correlated. Dark-adaptation did not change the number of ribbons and active zone puncta. However, mean ribbon length and length of presynaptic Cav1.4/RIM2 clusters increased significantly during dark-adaptation when tonic exocytosis is highest. In the present study, we identified by the analyses of synaptic ribbon-deficient RIBEYE knockout mice that synaptic ribbons are (1) needed to stabilize Cav1.4/RIM2 at rod photoreceptor active zones and (2) are required for the darkness-induced active zone enrichment of Cav1.4/RIM2. These data propose a role of the ribbon in active zone stabilization and suggest a homeostatic function of the ribbon in illumination-dependent active zone remodeling. The active zone of the presynaptic terminal controls central aspects of presynaptic communication 1,2. It is a protein-rich, electron-dense specialization of the plasma membrane at which synaptic vesicle fusion preferentially occurs. The active zone is strongly enriched in voltage-gated Cav-channels and active zone proteins, e.g. RIM-proteins, that attach the vesicle fusion machinery in close proximity to the Cav-channels for tight coupling creating rapidly reacting nanodomains 1,3-9. Ribbon synapses are continuously active chemical synapses with specialized active zones that are considered to promote both fast, transient and slow, continuous synaptic vesicle exocytosis 10,11. The active zone of ribbon synapses is extended by large electron-dense structures, synaptic ribbons, that recruit additional vesicles to the active zone. The basal row of ribbon-associated vesicles is positioned close to the Cav-channels and represents the fastest releasable vesicle pool 12 ideally suited to signal the onset of a stimulus. Synaptic vesicles tethered to the ribbon body, i.e. in some distance from the active zone, are considered to replenish empty release sites at the active zone with slower release kinetics providing information predominantly about the length of the stimulus 12,13. The main structural component of ribbons is the ribbon-specific protein RIBEYE 14,15. Photoreceptor ribbon synapses are the first synapses in the visual system at which light signals are transmitted to the inner retina. Light induces hyperpolarization of photoreceptors and a decrease of exocytosis at the synapse 16. Rod synapses represent the majority of photoreceptor synapses in the mouse retina and are built in a morphologically fairly uniform manner. They use a single large active zone with a single large ribbon. In EM cross-sections, rod ribbons typically ap...

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RIBEYE B-Domain Is Essential for RIBEYE A-Domain Stability and Assembly of Synaptic Ribbons

Shankhwar

Schwarz

Katiyar

et al. 2022

Front. Mol. Neurosci.

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Synaptic ribbons are presynaptic specializations that define eponymous ribbon synapses. Synaptic ribbons are largely composed of RIBEYE, a protein containing an N-terminal A-domain and a carboxyterminal B-domain that is identical with CtBP2, a NAD(H)-binding transcriptional co-repressor. Previously we showed that synaptic ribbons are completely absent in RIBEYE knockout mice in which the RIBEYE A-domain-encoding exon had been deleted, but CtBP2 is still made, demonstrating that the A-domain is required for synaptic ribbon assembly. In the present study, we asked whether the RIBEYE B-domain also has an essential role in the assembly of synaptic ribbons. For this purpose, we made use of RIBEYE knockin mice in which the RIBEYE B-domain was replaced by a fluorescent protein domain, whereas the RIBEYE A-domain was retained unchanged. We found that replacing the RIBEYE B-domain with a fluorescent protein module destabilizes the resulting hybrid protein and causes a complete loss of synaptic ribbons. Our results thus demonstrate an essential role of the RIBEYE B-domain in enabling RIBEYE assembly into synaptic ribbons, reinforcing the notion that RIBEYE is the central organizer of synaptic ribbons.

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The Calcineurin-Binding, Activity-Dependent Splice Variant Dynamin1xb Is Highly Enriched in Synapses in Various Regions of the Central Nervous System

Cited by 7 publications

References 136 publications

Early auto‐immune targeting of photoreceptor ribbon synapses in mouse models of multiple sclerosis

Early auto‐immune targeting of photoreceptor ribbon synapses in mouse models of multiple sclerosis

Synaptic ribbons foster active zone stability and illumination-dependent active zone enrichment of RIM2 and Cav1.4 in photoreceptor synapses

RIBEYE B-Domain Is Essential for RIBEYE A-Domain Stability and Assembly of Synaptic Ribbons

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