Syncoilin modulates peripherin filament networks and is necessary for large-calibre motor neurons

Clarke, William T; Edwards, Benjamin; McCullagh, Karl J. A.; Kemp, Matthew W.; Moorwood, Catherine; Sherman, Diane L.; Burgess, Matthew; Davies, Kay E.

doi:10.1242/jcs.059113

Cited by 22 publications

(14 citation statements)

References 44 publications

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“…Triple immunofluorescence labelling of Cx36, vglut1, and the motoneuron marker peripherin (Clarke et al, 2010) in lamina IX of adult rat spinal cord is shown at the L4 level (Fig. 2).…”

Section: Resultsmentioning

confidence: 98%

“…In addition, we used a chicken polyclonal anti-peripherin antibody (obtained from Millipore and diluted of 1:500) to identify motoneurons and primary afferent fibers in the spinal cord. Peripherin is an intermediate filament protein highly expressed in neurons of the peripheral nervous system, but is also found in central neurons that have projections to peripheral structures, including motoneurons which display intense labelling for this protein (Clarke et al, 2010). Various secondary antibodies included Cy3-conjugated goat or donkey anti-mouse and anti-rabbit IgG (Jackson ImmunoResearch Laboratories, West Grove, PA, USA), Alexa Flour 488-conjugated goat or donkey anti-rabbit and anti-mouse IgG (Molecular Probes, Eugene, OR, USA), AlexaFluor-647 conjugated goat anti-chicken IgG (Life Technologies Corporation) and Cy3-conjugated goat anti-chicken (Jackson ImmunoResearch Laboratories).…”

Section: Methodsmentioning

confidence: 99%

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Connexin36 identified at morphologically mixed chemical/electrical synapses on trigeminal motoneurons and at primary afferent terminals on spinal cord neurons in adult mouse and rat

2014

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Morphologically mixed chemical/electrical synapses at axon terminals, with the electrical component formed by gap junctions, is common in the CNS of lower vertebrates. In mammalian CNS, evidence for morphologically mixed synapses has been obtained in only a few locations. Here, we used immunofluorescence approaches to examine the localization of the neuronally expressed gap junction forming protein connexin36 (Cx36) in relation to the axon terminal marker vesicular glutamate transporter1 (vglut1) in spinal cord and trigeminal motor nucleus (Mo5) of rat and mouse. In adult rodents, immunolabelling for Cx36 appeared exclusively as Cx36-puncta, and was widely distributed at all rostro-caudal levels in most spinal cord laminae and in the Mo5. A high proportion of Cx36-puncta was co-localized with vglut1, forming morphologically mixed synapses on motoneurons, in intermediate spinal cord lamina, and in regions of medial lamina VII, where vglut1-containing terminals associated with Cx36 converged on neurons adjacent to the central canal. Unilateral transection of lumbar dorsal roots reduced immunolabelling of both vglut1 and Cx36 in intermediate laminae and lamina IX. Further, vglut1-terminals displaying Cx36-puncta were contacted by terminals labelled for glutamic acid decarboxylase65, which is known to be contained in presynaptic terminals on large diameter primary afferents. Developmentally, mixed synapses begin to emerge in the spinal cord only after the second to third postnatal week and thereafter increase to adult levels. Our findings demonstrate that axon terminals of primary afferent origin form morphologically mixed synapses containing Cx36 in broadly distributed areas of adult rodent spinal cord and Mo5.

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“…Triple immunofluorescence labelling of Cx36, vglut1, and the motoneuron marker peripherin (Clarke et al, 2010) in lamina IX of adult rat spinal cord is shown at the L4 level (Fig. 2).…”

Section: Resultsmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 99%

Connexin36 identified at morphologically mixed chemical/electrical synapses on trigeminal motoneurons and at primary afferent terminals on spinal cord neurons in adult mouse and rat

2014

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“…As shown in Fig. 6, peripherin displays an extensive staining of the neuronal network, which is expected from its established role in the neuronal cytoskeleton [32]. In contrast, RAB7A antibodies label punctate structures widely distributed in the neuronal network and in the cell body (Fig.…”

Section: Resultsmentioning

confidence: 99%

Charcot–Marie–Tooth type 2B disease-causing RAB7A mutant proteins show altered interaction with the neuronal intermediate filament peripherin

et al. 2012

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Charcot–Marie–Tooth type 2B (CMT2B) is a peripheral ulcero-mutilating neuropathy caused by four missense mutations in the rab7a gene. CMT2B is clinically characterized by prominent sensory loss, distal muscle weakness leading to muscle atrophy, high frequency of foot ulcers and infections that often results in toe amputations. RAB7A is a ubiquitous small GTPase, which controls transport to late endocytic compartments. Although the biochemical and functional properties of disease-causing RAB7A mutant proteins have been investigated, it is not yet clear how the disease originates. To understand how mutations in a ubiquitous protein specifically affect peripheral neurons, we performed a two-hybrid screen using a dorsal root ganglia cDNA library with the purpose of identifying RAB7A interactors specific for these cells. We identified peripherin, an intermediate filament protein expressed primarily in peripheral neurons, as a putative RAB7A interacting protein. The interaction was confirmed by co-immunoprecipitation and pull-down experiments, and established that the interaction is direct using recombinant proteins. Silencing or overexpression of wild type RAB7A changed the soluble/insoluble rate of peripherin indicating that RAB7A is important for peripherin organization and function. In addition, disease-causing RAB7A mutant proteins bind more strongly to peripherin and their expression causes a significant increase in the amount of soluble peripherin. Since peripherin plays a role not only in neurite outgrowth during development but also in axonal regeneration after injury, these data suggest that the altered interaction between disease-causing RAB7A mutants and peripherin could play an important role in CMT2B neuropathy.

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“…Total RNA (1.0 μg) was reverse transcribed using PrimeScript II 1st strand cDNA Synthesis Kit and SYBR green-based qRT-PCR was performed by using SYBR Select Master Mix (4472908; Life Technologies). Primers used were previously described, [26][27][28] and they are listed in Table 3.…”

Section: Quantitative Reverse Transcriptase-pcr (Qrt-pcr) Analysismentioning

confidence: 99%

Microangiopathy triggers, and inducible nitric oxide synthase exacerbates dextran sulfate sodium-induced colitis

Saijo

Tatsumi

Arihiro

et al. 2015

Laboratory Investigation

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Ulcerative colitis (UC) is a representative clinical manifestation of inflammatory bowel disease that causes chronic gastrointestinal tract inflammation. Dextran sulfate sodium (DSS)-induced colitis mice have been used to investigate UC pathogenesis, and in this UC model, disturbance and impairment of the mucosal epithelium have been reported to cause colitis. However, how DSS sporadically breaks down the epithelium remains unclear. In this study, we focused on the colonic microcirculation and myenteric neurons of DSS-induced colitis. Moreover, we examined the potential of myenteric neurons as a target to prevent exacerbation of colitis. Fluorescent angiographic and histopathological studies revealed that DSS administration elicited blood vessel disruption before epithelial disorders appeared. Ischemic conditions in the lamina propria induced inducible nitric oxide synthase (iNOS) expression in myenteric neurons as colitis aggravated. When neuronal activity was inhibited with butylscopolamine, neuronal iNOS expression decreased, and the exacerbation of colitis was prevented. These results suggested that DSS-induced colitis was triggered by microcirculatory disturbance in the mucosa, and that excessive neuronal excitation aggravated colitis. During remission periods of human UC, endoscopic inspection of the colonic microcirculation may enable the early detection of disease recurrence, and inhibition of neuronal iNOS expression may prevent the disease from worsening.

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Syncoilin modulates peripherin filament networks and is necessary for large-calibre motor neurons

Cited by 22 publications

References 44 publications

Connexin36 identified at morphologically mixed chemical/electrical synapses on trigeminal motoneurons and at primary afferent terminals on spinal cord neurons in adult mouse and rat

Connexin36 identified at morphologically mixed chemical/electrical synapses on trigeminal motoneurons and at primary afferent terminals on spinal cord neurons in adult mouse and rat

Charcot–Marie–Tooth type 2B disease-causing RAB7A mutant proteins show altered interaction with the neuronal intermediate filament peripherin

Microangiopathy triggers, and inducible nitric oxide synthase exacerbates dextran sulfate sodium-induced colitis

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