myelinated fibers
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2021 ◽  
Vol 118 (45) ◽  
pp. e2106143118
Author(s):  
Ming-Yu Wu ◽  
Alex Y. H. Wong ◽  
Jong-Kai Leung ◽  
Chuen Kam ◽  
Kenneth Lap-Kei Wu ◽  
...  

Myelin, the structure that surrounds and insulates neuronal axons, is an important component of the central nervous system. The visualization of the myelinated fibers in brain tissues can largely facilitate the diagnosis of myelin-related diseases and understand how the brain functions. However, the most widely used fluorescent probes for myelin visualization, such as Vybrant DiD and FluoroMyelin, have strong background staining, low-staining contrast, and low brightness. These drawbacks may originate from their self-quenching properties and greatly limit their applications in three-dimensional (3D) imaging and myelin tracing. Chemical probes for the fluorescence imaging of myelin in 3D, especially in optically cleared tissue, are highly desirable but rarely reported. We herein developed a near-infrared aggregation-induced emission (AIE)-active probe, PM-ML, for high-performance myelin imaging. PM-ML is plasma membrane targeting with good photostability. It could specifically label myelinated fibers in teased sciatic nerves and mouse brain tissues with a high–signal-to-background ratio. PM-ML could be used for 3D visualization of myelin sheaths, myelinated fibers, and fascicles with high-penetration depth. The staining is compatible with different brain tissue–clearing methods, such as ClearT and ClearT2. The utility of PM-ML staining in demyelinating disease studies was demonstrated using the mouse model of multiple sclerosis. Together, this work provides an important tool for high-quality myelin visualization across scales, which may greatly contribute to the study of myelin-related diseases.



2021 ◽  
Vol 15 ◽  
Author(s):  
Gerrit E. Gmel ◽  
Paul F. Vollebregt ◽  
Marjolein E. G. Thijssen ◽  
Rosana Santos Escapa ◽  
Eleanor McAlees ◽  
...  

Intra-operative electrode placement for sacral neuromodulation (SNM) relies on visual observation of motor contractions alone, lacking complete information on neural activation from stimulation. This study aimed to determine whether electrophysiological responses can be recorded directly from the S3 sacral nerve during therapeutic SNM in patients with fecal incontinence, and to characterize such responses in order to better understand the mechanism of action (MOA) and whether stimulation is subject to changes in posture. Eleven patients undergoing SNM were prospectively recruited. A bespoke stimulating and recording system was connected (both intraoperatively and postoperatively) to externalized SNM leads, and electrophysiological responses to monopolar current sweeps on each electrode were recorded and analyzed. The nature and thresholds of muscle contractions (intraoperatively) and patient-reported stimulation perception were recorded. We identified both neural responses (evoked compound action potentials) as well as myoelectric responses (far-field potentials from muscle activation). We identified large myelinated fibers (conduction velocity: 36–60 m/s) in 5/11 patients, correlating with patient-reported stimulation perception, and smaller myelinated fibers (conduction velocity <15 m/s) in 4/11 patients (not associated with any sensation). Myoelectric responses (observed in 7/11 patients) were attributed to pelvic floor and/or anal sphincter contraction. Responses varied with changes in posture. We present the first direct electrophysiological responses recorded from the S3 nerve during ongoing SNM in humans, showing both neural and myoelectric responses. These recordings highlight heterogeneity of neural and myoelectric responses (relevant to understanding MOA of SNM) and confirm that electrode lead position can change with posture.



2021 ◽  
Vol 10 (3) ◽  
pp. 83-90
Author(s):  
N. A. Shchudlo ◽  
T. N. Varsegova ◽  
M. M. Shchudlo

The effect of ageing on the morphofunctional features of nerves and the process of their posttraumatic regeneration has been studied in details. However, it is not known, whether there are differences of the neuroregeneration potential in the young, adult and mature experimental animals.The aim of the study was to evaluate histomorphometrical parameters of the rat sciatic nerve regeneration after cutting and microsurgical suturing depending on their age.Materials and Methods. The study included 15 white laboratory Wistar rats aged 5–6 (young), 8–10 (adult) and 12 months (mature). The sciatic nerve was transected at the level of the middle third of the thigh with microsurgical scissors and sutured with six epi-perineural stitches in the animals. In 4 months the animals were euthanized. Sections of sciatic nerves distal to the levels of sutures were put into araldite, semi-thin slices (0.5–1.0 mkm) were obtained for histomorphometric analysis. Thirteen intact animals were selected as control. Statistical processing was performed using nonparametric methods.Results. The numerical density of endoneural vessels in nerves of operated rats was higher than that in the intact ones, but in mature animals it was significantly lower than in young and adult ones (p<0.001). The numerical density of endoneural cells nuclei in young experimental animals did not differ from that of the intact ones, but in animals from the adult and mature groups it was more than four times higher than in animals from the young group (p<0.001). The numerical density of regenerating myelinated nerve fibers in animals of the young and adult groups did not differ from that of the intact nerve, in animals of the mature group it increased in 2.5 times (p<0.001). The numerical density of degenerating myelinated fibers in the operated animals of experimental groups was higher than in the intact group (two times, six times and 11 times higher, respectively). The diameter distribution of the regenerating myelinated fibers in animals of all groups differed from that of the intact control animals.Conclusions. The data obtained demonstrate a significant decrease in the regenerative potential of Schwann cells and neurons in adult and mature animals compared with the young ones.



Glia ◽  
2021 ◽  
Author(s):  
Eriola Hoxha ◽  
Ilaria Balbo ◽  
Roberta Parolisi ◽  
Matteo Audano ◽  
Francesca Montarolo ◽  
...  


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Irene Costantini ◽  
Enrico Baria ◽  
Michele Sorelli ◽  
Felix Matuschke ◽  
Francesco Giardini ◽  
...  

AbstractAnalyzing the structure of neuronal fibers with single axon resolution in large volumes is a challenge in connectomics. Different technologies try to address this goal; however, they are limited either by the ineffective labeling of the fibers or in the achievable resolution. The possibility of discriminating between different adjacent myelinated axons gives the opportunity of providing more information about the fiber composition and architecture within a specific area. Here, we propose MAGIC (Myelin Autofluorescence imaging by Glycerol Induced Contrast enhancement), a tissue preparation method to perform label-free fluorescence imaging of myelinated fibers that is user friendly and easy to handle. We exploit the high axial and radial resolution of two-photon fluorescence microscopy (TPFM) optical sectioning to decipher the mixture of various fiber orientations within the sample of interest. We demonstrate its broad applicability by performing mesoscopic reconstruction at a sub-micron resolution of mouse, rat, monkey, and human brain samples and by quantifying the different fiber organization in control and Reeler mouse's hippocampal sections. Our study provides a novel method for 3D label-free imaging of nerve fibers in fixed samples at high resolution, below micrometer level, that overcomes the limitation related to the myelinated axons exogenous labeling, improving the possibility of analyzing brain connectivity.



2021 ◽  
Vol 12 (2) ◽  
pp. 73-91
Author(s):  
Taras Kotyk ◽  
Nadiya Tokaruk ◽  
Viktoria Bedej ◽  
Mariia Hryshchuk ◽  
Oksana Popadynets ◽  
...  

One of the unresolved issues in experimental neuromorphology is searching for a solution for myelinated nerve fibers clustering on set of morphometric parameters. Therefore, in this article, a new approach for cluster analysis of myelinated fibers is proposed based on their morpho-functional features. The proposed clustering approach was developed in R software environment and uses model-based clustering, which is performed in few steps with increasing number of morphometric parameters on each next step. Applying the proposed clustering solution shown high similarity of identified groups' morphometric parameters with respective physiological types of myelinated A-fibers. This fact, in addition to the algorithm implementation simplicity, facilitates its use on identifying clusters of myelinated fibers that represent different myelinated fibers subpopulation in experimental neuromorphological research with high level of reliability.



2021 ◽  
Vol 10 (4) ◽  
pp. 824
Author(s):  
Daisuke Maki ◽  
Tetsuro Tamaki ◽  
Tsuyoshi Fukuzawa ◽  
Toshiharu Natsume ◽  
Ippei Yamato ◽  
...  

Severe peripheral nerve injury, which does not promise natural healing, inevitably requires clinical treatment. Here, we demonstrated the facilitation effect of peripheral nerve regeneration using a cytokine cocktail secreted by skeletal muscle-derived stem cells (Sk-MSCs). Mouse sciatic nerve was transected with a 6 mm gap and bridged collagen tube, and the culture supernatant of Sk-MSCs with 20% adult mouse serum (AMS)/Iscove’s modified Dulbecco’s medium (IMDM) was administered into the tube immediately after the operation, followed by an injection once a week for six weeks through the skin to the surrounding tube of the cytokine (CT) group. Similarly, 20% AMS/IMDM without cytokines was administered to the non-cytokine control (NT) group. Tension recovery in the plantar flexor muscles via electrical stimulation at the upper portion of the damaged nerve site, as well as the numerical recovery of axons and myelinated fibers at the damaged site, were evaluated as an index of nerve regeneration. Specific cytokines secreted by Sk-MSCs were compared with damaged sciatic nerve-derived cytokines. Six weeks after operation, significantly higher tension output and numerical recovery of the axon and myelinated fibers were consistently observed in the CT group, showing that the present cytokine cocktail may be a useful nerve regeneration acceleration agent. We also determined 17 candidate factors, which are likely included in the cocktail.



Life ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 136
Author(s):  
Wiebke Möbius ◽  
Sophie Hümmert ◽  
Torben Ruhwedel ◽  
Alan Kuzirian ◽  
Robert Gould

Although myelinated nervous systems are shared among 60,000 jawed vertebrates, studies aimed at understanding myelination have focused more and more on mice and zebrafish. To obtain a broader understanding of the myelination process, we examined the little skate, Leucoraja erinacea. The reasons behind initiating studies at this time include: the desire to study a species belonging to an out group of other jawed vertebrates; using a species with embryos accessible throughout development; the availability of genome sequences; and the likelihood that mammalian antibodies recognize homologs in the chosen species. We report that the morphological features of myelination in a skate hatchling, a stage that supports complex behavioral repertoires needed for survival, are highly similar in terms of: appearances of myelinating oligodendrocytes (CNS) and Schwann cells (PNS); the way their levels of myelination conform to axon caliber; and their identity in terms of nodal and paranodal specializations. These features provide a core for further studies to determine: axon–myelinating cell communication; the structures of the proteins and lipids upon which myelinated fibers are formed; the pathways used to transport these molecules to sites of myelin assembly and maintenance; and the gene regulatory networks that control their expressions.



2020 ◽  
Vol 10 (12) ◽  
pp. 989
Author(s):  
Maria Nolano ◽  
Stefano Tozza ◽  
Giuseppe Caporaso ◽  
Vincenzo Provitera

In the last three decades the study of cutaneous innervation through 3 mm-punch-biopsy has provided an important contribution to the knowledge of small fiber somatic and autonomic neuropathies but also of large fiber neuropathies. Skin biopsy is a minimally invasive technique with the advantage, compared to sural nerve biopsy, of being suitable to be applied to any site in our body, of being repeatable over time, of allowing the identification of each population of nerve fiber through its target. In patients with symptoms and signs of small fiber neuropathy the assessment of IntraEpidermal Nerve Fiber density is the gold standard to confirm the diagnosis while the quantification of sudomotor, pilomotor, and vasomotor nerve fibers allows to evaluate and characterize the autonomic involvement. All these parameters can be re-evaluated over time to monitor the disease process and to evaluate the effectiveness of the treatments. Myelinated fibers and their receptors can also be evaluated to detect a “dying back” neuropathy early when nerve conduction study is still normal. Furthermore, the morphometry of dermal myelinated fibers has provided new insight into pathophysiological mechanisms of different types of inherited and acquired large fibers neuropathies. In genetic neuropathies skin biopsy has become a surrogate for sural nerve biopsy, no longer necessary in the diagnostic process, to study genotype–phenotype correlations.



2020 ◽  
Author(s):  
Irene Costantini ◽  
Enrico Baria ◽  
Michele Sorelli ◽  
Felix Matuschke ◽  
Francesco Giardini ◽  
...  

AbstractAnalyzing the structure of neuronal fibers with single axon resolution, in large volumes, remains an unresolved challenge in connectomics. Here, we propose MAGIC (Myelin Autofluorescence imaging by Glycerol Induced Contrast enhancement), a simple tissue preparation method to perform label-free fluorescence imaging of myelinated fibers. We demonstrate its broad applicability by performing mesoscopic reconstruction at sub-micron resolution of mouse, rat, monkey, and human brain samples and by quantifying the different fiber organization in Control and Reeler mouse’s hippocampal sections.



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