Developmental trajectory of oligodendrocyte progenitor cells in the human brain revealed by single cell RNA sequencing.

Characterizing the developmental trajectory of oligodendrocyte progenitor cells (OPC) is of great interest given the importance of these cells in the remyelination process. However, studies of human OPC development remain limited by the availability of whole cell samples and material that encompasses a wide age range, including time of peak myelination. In this study, we apply single cell RNA sequencing to viable whole cells across the age span and link transcriptomic signatures of oligodendrocyte-lineage cells with stage-specific functional properties. Cells were isolated from surgical tissue samples of second-trimester fetal, 2-year-old pediatric, 13-year-old adolescent, and adult donors by mechanical and enzymatic digestion, followed by percoll gradient centrifugation. Gene expression was analyzed using droplet-based RNA sequencing (10X Chromium). Louvain clustering analysis identified three distinct cellular subpopulations based on 5,613 genes, comprised of an early OPC (e-OPC) group, a late OPC group (l-OPC), and a mature OL (MOL) group. Gene ontology terms enriched for e-OPCs included cell cycle and development, for l-OPCs included extracellular matrix and cell adhesion, and for MOLs included myelination and cytoskeleton. The e-OPCs were mostly confined to the premyelinating fetal group, and the l-OPCs were most highly represented in the pediatric age group, corresponding to the peak age of myelination. Cells expressing a signature characteristic of l-OPCs were identified in the adult brain in situ using RNAScope. These findings highlight the transcriptomic variability in OL-lineage cells before, during, and after peak myelination and contribute to identifying novel pathways required to achieve remyelination.

Microsurgical anatomy of lumbosacral nerve rootlets for highly selective rhizotomy in chronic spinal cord injury.

It is known that selective sacral roots rhizotomy is effective for relieving the neurogenic bladder associated with spinal cord injury. The goal of this study is to review the surgical anatomy of the lumbosacral nerve rootlets and to provide some morphological bases for highly selective sacral roots rhizotomy. Spinal cord dissections were performed on five cadavers under surgical microscope. At each spinal cord segment, we recorded the number, diameter and length of the rootlets, subbundles and bundles from the L1 to S2 spinal segments, and the length of the dorsal/ventral root entry zone. Peripheral nervous system myelin was examined by immunohistochemistry. We found: (1) the ventral or the dorsal root of the lumbosacral segment of the spinal cord was divided into one to three nerve bundles and each bundle was subdivided into one to three subbundles. Each subbundle further gave out two to three rootlets connected with the spinal cord; (2) there were no significant differences in the number of rootlets within the L1 to S2 segments, but the size of rootlets and the length of nerve roots varied (P < 0.05); and (3) the more myelinated fibers a rootlet contained, the larger transection area it had. The area of peripheral nervous system myelin positive cells and the total area of rootlets were correlated (P < 0.001). Thus, during highly selective sacral roots rhizotomy, the ventral and dorsal roots can be divided into several bundles of rootlets, and we could initially distinct the rootlets by their diameters.

Uncompacted myelin lamellae in peripheral nerve biopsy.

Since 1979, the authors have studied 49 peripheral nerve biopsies presenting uncompacted myelin lamellae (UML). Based on the ultrastructural pattern of UML they propose a 3-category classification. The first category includes cases displaying regular UML, which was observed in 43 cases; it was more frequent in 9 cases with polyneuropathy organomegaly endocrinopathy m-protein skin changes (POEMS) syndrome as well as in 1 case of Charcot-Marie-Tooth 1B with a novel point mutation in the P0 gene. The second category consists of cases showing irregular UML, observed in 4 cases with IgM monoclonal gammopathy and anti-myelin-associated glycoprotein (MAG) activity. This group included 1 benign case and 3 B-cell malignant lymphomas. The third category is complex UML, which was present in 2 unrelated patients with an Arg 98 His missense mutation in the P0 protein gene. Irregular and complex UML are respectively related to MAG and P0, which play a crucial role in myelin lamellae compaction and adhesion.