Dorsal Horn of Mouse Lumbar Spinal Cord Imaged with CLARITY.

The organization of the mouse spinal dorsal horn has been delineated in 2D for the six Rexed laminae in our publication Atlas of the Spinal Cord: Mouse, Rat, Rhesus, Marmoset, and Human. In the present study, the tissue clearing technique CLARITY was used to observe the cyto- and chemoarchitecture of the mouse spinal cord in 3D, using a variety of immunohistochemical markers. We confirm prior observations regarding the location of glycine and serotonin immunoreactivities. Novel observations include the demonstration of numerous calcitonin gene-related peptide (CGRP) perikarya, as well as CGRP fibers and terminals in all laminae of the dorsal horn. We also observed sparse choline acetyltransferase (ChAT) immunoreactivity in small perikarya and fibers and terminals in all dorsal horn laminae, while gamma aminobutyric acid (GABA) and glutamate decarboxylase-67 (GAD67) immunoreactivities were found only in small perikarya and fibers. Finally, numerous serotonergic fibers were observed in all laminae of the dorsal horn. In conclusion, CLARITY confirmed the 2D immunohistochemical properties of the spinal cord. Furthermore, we observed novel anatomical characteristics of the spinal cord and demonstrated that CLARITY can be used on spinal cord tissue to examine many proteins of interest.

Normal tolerance limits for side-to-side differences in diameters of major lower limbs arteries of 228 healthy subjects.

PURPOSE: To determine reference values and tolerance limits of between-side differences for the calibers of the common femoral artery (CFA), superficial femoral artery (SFA), popliteal artery (PA), dorsalis pedis artery (DPA), and posterior tibial artery (PTA). MATERIALS AND METHODS: Calibers of arteries, defined as the largest distance between internal hyperechogenic lines of the intima-media complex of the arterial wall, were measured during the diastole phase determined from echo-tracking B mode ultrasound scanning and grey-scale ultrasound in 228 healthy volunteers aged 18-81 years (43.1 ± 16.7). RESULTS: The mean, 95% confidence and tolerance limits covering 90% of population for left and right side of each artery were: CFA: 8.1 mm, 7.9-8.3 mm, 6.0-10.3 mm; 8.1 mm, 7.9-8.5 mm, 5.9-10.2 mm; SFA: 6.2 mm, 6.0-6.3 mm, 4.7-7.6 mm; 6.1 mm, 6.0-6.3 mm, 4.7-7.6 mm; PA: 6.1 mm, 6.0-6.2 mm, 4.6-7.6 mm; 6.1 mm, 5.9-6.2 mm, 4.5-7.6 mm; DPA: 2.0 mm, 1.9-2.0 mm, 1.2-2.7 mm; 2.0 mm, 1.9-2.0 mm, 1.2-2.8 mm; PTA: 2.1 mm, 2.0-2.1 mm, 1.4-2.8 mm; 2.1 mm, 2.1-2.2 mm, 1.4-2.8 mm, respectively. Tolerance limits for between-side differences and ratios were: CFA - 0.5-0.7 mm, 0.9-1.1; SFA - 0.5-0.6 mm, 0.9-1.1; PA - 0.5-0.5 mm, 0.9-1.1; DPA -0.4-0.4 mm, 0.8-1.2; PTA - 0.4-0.4 mm, 0.8-1.2. Regression analysis showed weight and age dependency of vessels diameters. There are no differences between men and woman in vessels size, except in DPA's, when body weight and age are taken into account in a regression analysis. CONCLUSIONS: We estimated normal reference tolerance limits of side-to-side differences in diameters of lower limb arteries. The limits can inform an investigator what differences in diameters occur in healthy individuals, and hence can serve as cut-offs in diagnostic and screening strategies.

MRI characteristics for "phantom" scratching in canine syringomyelia.

BACKGROUND: A classic sign of canine syringomyelia (SM) is scratching towards one shoulder. Using magnetic resonance imaging (MRI) we investigate the spinal cord lesion relating to this phenomenon which has characteristics similar to fictive scratch secondary to spinal cord transection. Medical records were searched for Cavalier King Charles spaniels with a clinical and MRI diagnosis of symptomatic SM associated with Chiari-like malformation (CM). The cohort was divided into SM with phantom scratching (19 dogs) and SM but no phantom scratching (18 dogs). MRI files were anonymised, randomised and viewed in EFILM ™. For each transverse image, the maximum perpendicular dimensions of the syrinx in the dorsal spinal cord quadrants were determined. Visual assessment was made as to whether the syrinx extended to the superficial dorsal horn (SDH). RESULTS: We showed that phantom scratching appears associated with a large dorsolateral syrinx that extends to the SDH in the C3-C6 spinal cord segments (corresponding to C2-C5 vertebrae). Estimated dorsal quadrant syrinx sizes based on the perpendicular diameters were between 2.5 and 9.5 times larger in dogs with phantom scratching, with the largest mean difference p-value being 0.009. CONCLUSION: SM associated phantom scratching appears associated with MRI findings of a large syrinx extending into the mid cervical SDH. We hypothesise that damage in this region might influence the lumbosacral scratching central pattern generator (CPG). If a scratching SM affected dog does not have a large dorsolateral cervical syrinx with SDH involvement then alternative explanations for scratching should be investigated.

Neurodegeneration in the Spinal Ventral Horn Prior to Motor Impairment in Cervical Spondylotic Myelopathy.

Remote gray matter pathology has been suggested rostral to the compression site in cervical spondylotic myelopathy (CSM). We therefore assessed neurodegeneration in the gray matter ventral and dorsal horns. Twenty patients with CSM and 18 healthy subjects underwent a high-resolution structural and diffusion magnetic resonance imaging protocol at vertebra C2/C3. Patients received comprehensive clinical assessments. T2*-weighted data provided cross-sectional area measurements of gray matter ventral and dorsal horns to identify atrophy. At the identical location, mean diffusivity (MD) and fractional anisotropy (FA) determined the microstructural integrity. Finally, the relationships between neurodegeneration occurring in the gray and white matter and clinical impairment were investigated. Patients suffered from mild-to-moderate CSM with mainly sensory impairment. In the ventral horns, cross-sectional area was not reduced (p = 0.863) but MD was increased (p = 0.045). The magnitude of MD changes within the ventral horn was associated with white matter diffusivity changes (MD: p = 0.013; FA: p = 0.028) within the lateral corticospinal tract. In contrast, dorsal horn cross-sectional area was reduced by 16.0% (p < 0.001) without alterations in diffusivity indices, compared with controls. No associations between the magnitude of ventral and dorsal horn neurodegeneration and clinical impairment were evident. Focal cord gray matter pathology is evident remote to the compression site in vivo in CSM patients. Microstructural changes in the ventral horns (i.e., motoneurons) related to corticospinal tract integrity in the absence of atrophy and marked motor impairment. Dorsal horn atrophy corresponded to main clinical representation of sensory impairment. Thus, neuroimaging biomarkers of cord gray matter integrity reveal focal neurodegeneration prior to marked clinical impairment and thus could serve as predictors of ensuing impairment in CSM patients.

Investigating resting-state functional connectivity in the cervical spinal cord at 3T.

The study of spontaneous fluctuations in the blood-oxygen-level-dependent (BOLD) signal has recently been extended from the brain to the spinal cord. Two ultra-high field functional magnetic resonance imaging (fMRI) studies in humans have provided evidence for reproducible resting-state connectivity between the dorsal horns as well as between the ventral horns, and a study in non-human primates has shown that these resting-state signals are impacted by spinal cord injury. As these studies were carried out at ultra-high field strengths using region-of-interest (ROI) based analyses, we investigated whether such resting-state signals could also be observed at the clinically more prevalent field strength of 3T. In a reanalysis of a sample of 20 healthy human participants who underwent a resting-state fMRI acquisition of the cervical spinal cord, we were able to observe significant dorsal horn connectivity as well as ventral horn connectivity, but no consistent effects for connectivity between dorsal and ventral horns, thus replicating the human 7T results. These effects were not only observable when averaging along the acquired length of the spinal cord, but also when we examined each of the acquired spinal segments separately, which showed similar patterns of connectivity. Finally, we investigated the robustness of these resting-state signals against variations in the analysis pipeline by varying the type of ROI creation, temporal filtering, nuisance regression and connectivity metric. We observed that - apart from the effects of band-pass filtering - ventral horn connectivity showed excellent robustness, whereas dorsal horn connectivity showed moderate robustness. Together, our results provide evidence that spinal cord resting-state connectivity is a robust and spatially consistent phenomenon that could be a valuable tool for investigating the effects of pathology, disease progression, and treatment response in neurological conditions with a spinal component, such as spinal cord injury.