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Myelin sheath formation in the peripheral nervous system and the ensuing saltatory conduction rely on differentiated Schwann cells. We have previously shown that transition of Schwann cells from an immature into a differentiated state requires Brg1 that serves as the central energy generating subunit in two related SWI/SNF-type chromatin remodelers, the BAF and the PBAF complex. Here we used conditional deletion of Pbrm1 to selectively interfere with the PBAF complex in Schwann cells. Despite efficient loss of Pbrm1 early during lineage progression, we failed to detect any substantial alterations in the number, proliferation or survival of immature Schwann cells as well as in their rate and timing of terminal differentiation. As a consequence, postnatal myelin formation in peripheral nerves appeared normal. There were no inflammatory alterations in the nerve or other signs of a peripheral neuropathy. We conclude from our study that Pbrm1 and very likely the PBAF complex are dispensable for proper Schwann cell development and that Schwann cell defects previously observed upon Brg1 deletion are mostly attributable to altered or absent function of the BAF complex., (© 2022. The Author(s).)

The three SoxD proteins, Sox5, Sox6 and Sox13, represent closely related transcription factors with important roles during development. In the developing nervous system, SoxD proteins have so far been primarily studied in oligodendroglial cells and in interneurons of brain and spinal cord. In oligodendroglial cells, Sox5 and Sox6 jointly maintain the precursor state, interfere with terminal differentiation, and thereby ensure the proper timing of myelination in the central nervous system. Here we studied the role of SoxD proteins in Schwann cells, the functional counterpart of oligodendrocytes in the peripheral nervous system. We show that Schwann cells express Sox5 and Sox13 but not Sox6. Expression was transient and ceased with the onset of terminal differentiation. In mice with early Schwann cell-specific deletion of both Sox5 and Sox13, embryonic Schwann cell development was not substantially affected and progressed normally into the promyelinating stage. However, there was a mild and transient delay in the myelination of the peripheral nervous system of these mice. We therefore conclude that SoxD proteins-in stark contrast to their action in oligodendrocytes-promote differentiation and myelination in Schwann cells.

Schwann cells are the nerve ensheathing cells of the peripheral nervous system. Absence, loss and malfunction of Schwann cells or their myelin sheaths lead to peripheral neuropathies such as Charcot-Marie-Tooth disease in humans. During Schwann cell development and myelination chromatin is dramatically modified. However, impact and functional relevance of these modifications are poorly understood. Here, we analyzed histone H2B monoubiquitination as one such chromatin modification by conditionally deleting the Rnf40 subunit of the responsible E3 ligase in mice. Rnf40-deficient Schwann cells were arrested immediately before myelination or generated abnormally thin, unstable myelin, resulting in a peripheral neuropathy characterized by hypomyelination and progressive axonal degeneration. By combining sequencing techniques with functional studies we show that H2B monoubiquitination does not influence global gene expression patterns, but instead ensures selective high expression of myelin and lipid biosynthesis genes and proper repression of immaturity genes. This requires the specific recruitment of the Rnf40-containing E3 ligase by Egr2, the central transcriptional regulator of peripheral myelination, to its target genes. Our study identifies histone ubiquitination as essential for Schwann cell myelination and unravels new disease-relevant links between chromatin modifications and transcription factors in the underlying regulatory network., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

The cause of the elevated outflow resistance and consequent ocular hypertension characteristic of glaucoma is unknown. To investigate possible causes for this flow resistance, we used atomic force microscopy (AFM) with 10-µm spherical tips to probe the stiffness of the inner wall of Schlemm's canal as a function of distance from the tissue surface in normal and glaucomatous postmortem human eyes, and 1-µm spherical AFM tips to probe the region immediately below the tissue surface. To localize flow resistance, perfusion and imaging methods were used to characterize the pressure drop in the immediate vicinity of the inner wall using giant vacuoles that form in Schlemm's canal cells as micropressure sensors. Tissue stiffness increased with increasing AFM indentation depth. Tissues from glaucomatous eyes were stiffer compared with normal eyes, with greatly increased stiffness residing within ∼1 µm of the inner-wall surface. Giant vacuole size and density were similar in normal and glaucomatous eyes despite lower flow rate through the latter due to their higher flow resistance. This implied that the elevated flow resistance found in the glaucomatous eyes was localized to the same region as the increased tissue stiffness. Our findings implicate pathological changes to biophysical characteristics of Schlemm's canal endothelia and/or their immediate underlying extracellular matrix as cause for ocular hypertension in glaucoma.

The ultrastructure of the nephrostomial tubule and the gonadal crest of Acipenser ruthenus were studied by transmission and scanning electron microscopy. Opisthonephric nephrostomial tubules begin to appear at the end of the first week after hatching and are regularly segmentally arranged and completely developed in the period between 10 and 35 days after hatching. Fully differentiated nephrostomial tubules connect the vestibular area of the Malpighian corpuscle with the coelomic cavity as short, open canals. Before reaching the latter, each nephrostomial tubule widens to a funnel-like structure and ends with a round opening, the nephrostome. The outer rim of the nephrostome protrudes slightly into the coelomic cavity forming the nephrostomial lips. The nephrostomial tubule is lined by one layer of cuboidal or low-columnar cells, equipped with an apical set of long kinocilia, which point generally in the direction of the Malpighian corpuscle. The cilia of the distal funnel region, however, point toward the coelomic cavity. The cells of the nephrostomial lips preserve a lower degree of cellular differentiation than the rest, display a blastema-like appearance and proliferate by frequent mitoses. Proliferating cells of the nephrostomial lips spread out on the coelomic surface and replace the flat mesothelium of lateral plate origin here. Furthermore, cells of the nephrostomial lips also show the tendency to grow downward, giving the epithelium of the lip region a multilayered appearance. Proliferating cells of the medial nephrostomial lips associate with accumulations of germ cells and form the primordium of the gonad (gonadal crest). Within the crest, the intercellular space between the germ cells and the surrounding epithelial supporting cells is filled with a basal lamina-like, electron-dense substance and may contain short, interlocking processes of the two cell types. The large germ cells of this early period have lobulated, electron-lucent nuclei, spherical mitochondria with inclusions and abundant narrow profiles of the smooth endoplasmatic reticulum.

Early gonadal development was studied in the sterlet, Acipenser ruthenus, by means of histological and semithin serial sections and scanning electron microscopy. Special attention was given to the role of opisthonephric nephrostomial tubules and their coelomic funnels (nephrostomes, coelomostomes) in the origin of the gonad. Specimens of about 1 mm in length (about 7 days post hatching) have a continuous kidney complex (holonephros) that extends from the branchial region to the level of the cloaca and may be divided into a cranial pronephros and a caudally following opisthonephros, with no overlapping of either portion. In specimens of 10 to 25 mm in length the regression of pronephros and cranial opisthonephros can already been seen; as a consequence, these parts of the kidney complex are not involved in gonadogenesis. The initial gonadal anlage is seen in specimens of 30-40 mm in length. The somatic cells of the gonadal primordium develop from the medial lips of segmentally arranged opisthonephric nephrostomes situated in a line that extends from the level of the stomach through that of the spiral valve. The nephrostomes involved in this process belong to the first-order set of nephrons, since nephrons of higher order that arise continuously from blastema cells during further growth of the animals never send nephrostomial tubules to the coelomic surface. The cells of the medial nephrostomial lips proliferate by many mitoses. They grow over and surround the large germ cells that have accumulated on the medial side of the nephrostomes. The proliferating nephrostomial cells are elongated in shape, and their long axes are oriented in the cranio-caudal direction. By their size, shape and arrangement they replace not only the flat, polygonal, mesothelial cells of lateral plate origin on the medial side of the nephrostomes, but also those in the interstices in between. The result is the formation of a continuous gonadal crest situated medially from the nephrostomial line. At 100-130 mm in length, the gonadal crest has reached the stage of a gonadal fold that is attached to the dorsal body wall by a thin mesogonadium. Stroma cells and blood vessels start to invade the gonadal fold. At 240-290 mm in length, parts of the gonadal fold are converted into a fat body, a structure that is typically present in adult sturgeons of either sex. Nephrostomial tubules and their nephrostomes are visible only for a short period in Acipenser ruthenus. They are completely developed in specimens of 25-40 mm in length and start continuous retrogression immediately afterwards. Therefore, they cannot play an important role in excretion. Their only purpose apparently is to function as precursor tissue for other organs, such as the gonad, in establishing a route by which cells of the intermediate mesoderm can gain access to and spread out over the coelomic surface. In conclusion, the observations made in Acipenser prove that this species is a most suitable model to explain the origin of the gonad from opistho/mesonephric nephrostomial tubules in vertebrates with a less-developed or early regressing pronephros. Most mammals, including man, belong to this category.