Loss of the Reissner Fiber and increased URP neuropeptide signaling underlie scoliosis in a zebrafish ciliopathy mutant

SUMMARYCilia-driven movements of the cerebrospinal fluid (CSF) are involved in zebrafish axis straightness, both in embryos and juveniles [1, 2]. In embryos, axis straightness requires cilia-dependent assembly of the Reissner fiber (RF), a SCO-spondin polymer running down the brain and spinal cord CSF-filled cavities [3]. Reduced expression levels of the urp1 and urp2 genes encoding neuropeptides of the Urotensin II family in CSF-contacting neurons (CSF-cNs) also underlie embryonic ventral curvature of several cilia motility mutants [4]. Moreover, mutants for scospondin and uts2r3 (a Urotensin II peptide family receptor gene) develop scoliosis at juvenile stages [3, 4]. However, whether RF maintenance and URP signaling are perturbed in juvenile scoliotic ciliary mutants and how these perturbations are linked to scoliosis is unknown. Here we produced mutants in the zebrafish ortholog of the human RPGRIP1L ciliopathy gene encoding a transition zone protein [5–7]. rpgrip1l-/- zebrafish had normal embryogenesis and developed 3D spine torsions in juveniles. Cilia lining the CNS cavities were normal in rpgrip1l-/- embryos but sparse and malformed in juveniles and adults. Hindbrain ventricle dilations were present at scoliosis onset, suggesting defects in CSF flow. Immunostaining showed a secondary loss of RF correlating with juvenile scoliosis. Surprisingly, transcriptome analysis of rgprip1l mutants at scoliosis onset uncovered increased levels of urp1 and urp2 expression. Overexpressing urp2 in foxj1-expressing cells triggered scoliosis in rpgrip1l heterozygotes. Thus, our results demonstrate that increased URP signaling drives scoliosis onset in a ciliopathy mutant. We propose that imbalanced levels of URP neuropeptides in CSF-cNs may be an initial trigger of scoliosis.

An Online Simultaneous Measurement of the Dual-Axis Straightness Error for Machine Tools

Vertical straightness errors are the key factor that affects the flatness of the workpiece during vertical machining. Traditionally, the individually measured and fitted vertical straightness errors of the X and Y axes are used to compensate the Z axis and, thus, obtain the flatness of the working table of the machine tool. However, it is difficult to measure and compensate the vertical straightness error of the desired position on the working table, not to mention the centroid variation effect of the working table on the measured data. In this study, an online dual-axis measurement system with repeatability (3σ) of 2.46 μm is developed to simultaneously measure X-axis and Y-axis straightness errors of the desired position of a working table. Furthermore, the measured data are utilized to establish a flatness error model to reduce the vertical straightness error of the working table such that the repeatability (3σ) of the measured flatness may be kept within a range of 0.65 μm.