Fabry-Pérot optical sensor and portable detector for monitoring high-resolution ocular hemodynamics.

Our understanding of ocular hemodynamics and its role in ophthalmic disease progression remains unclear due to the shortcomings of precise and on-demand biomedical sensing technologies. Here, we report high-resolution in vivo assessment of ocular hemodynamics using a Fabry-Pérot cavity-based micro-optical sensor and a portable optical detector. The designed optical system is capable of measuring both static intraocular pressure and dynamic ocular pulsation profiles in parallel. Through a dynamic intensity variation analysis method which improves sensing resolution by 3-4 folds, our system is able to extract systolic/diastolic phases from a single ocular pulsation profile. Using a portable detector, we performed in vivo studies on rabbits and verified that ophthalmic parameters obtained from our optical system closely match with traditional techniques such as tonometry, electrocardiography, and photo-plethysmography.

Real-Time In Vivo Intraocular Pressure Monitoring using an Optomechanical Implant and an Artificial Neural Network.

Optimized glaucoma therapy requires frequent monitoring and timely lowering of elevated intraocular pressure (IOP). A recently developed microscale IOP-monitoring implant, when illuminated with broadband light, reflects a pressure-dependent optical spectrum that is captured and converted to measure IOP. However, its accuracy is limited by background noise and the difficulty of modeling non-linear shifts of the spectra with respect to pressure changes. Using an end-to-end calibration system to train an artificial neural network (ANN) for signal demodulation we improved the speed and accuracy of pressure measurements obtained with an optically probed IOP-monitoring implant and make it suitable for real-time in vivo IOP monitoring. The ANN converts captured optical spectra into corresponding IOP levels. We achieved an IOP-measurement accuracy of ±0.1 mmHg at a measurement rate of 100 Hz, which represents a ten-fold improvement from previously reported values. This technique allowed real-time tracking of artificially induced sub-1 s transient IOP elevations and minor fluctuations induced by the respiratory motion of the rabbits during in vivo monitoring. All in vivo sensor readings paralleled those obtained concurrently using a commercial tonometer and showed consistency within ±2 mmHg. Real-time processing is highly useful for IOP monitoring in clinical settings and home environments and improves the overall practicality of the optical IOP-monitoring approach.

Ectopic vesicular glutamate release at the optic nerve head and axon loss in mouse experimental glaucoma.

Although clinical and experimental observations indicate that the optic nerve head (ONH) is a major site of axon degeneration in glaucoma, the mechanisms by which local retinal ganglion cell (RGC) axons are injured and damage spreads among axons remain poorly defined. Using a laser-induced ocular hypertension (LIOH) mouse model of glaucoma, we found that within 48 h of intraocular pressure elevation, RGC axon segments within the ONH exhibited ectopic accumulation and colocalization of multiple components of the glutamatergic presynaptic machinery including the vesicular glutamate transporter VGLUT2, several synaptic vesicle marker proteins, glutamate, the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex and active zone cytomatrix components, as well as ultrastructurally identified, synaptophysin-containing vesicles. Ectopic vesicle exocytosis and glutamate release were detected in acute preparations of the LIOH ONH. Immunolocalization and analysis using the ionotropic receptor channel-permeant cation agmatine indicated that ONH axon segments and glia expressed glutamate receptors, and these receptors were more active after LIOH compared with controls. Pharmacological antagonism of glutamate receptors and neuronal activity resulted in increased RGC axon sparing in vivo. Furthermore, in vivo RGC-specific genetic disruption of the vesicular glutamate transporter VGLUT2 or the obligatory NMDA receptor subunit NR1 promoted axon survival in experimental glaucoma. As the inhibition of ectopic glutamate vesicular release or glutamate receptivity can independently modify the severity of RGC axon loss, synaptic release mechanisms may provide useful therapeutic entry points into glaucomatous axon degeneration.

Automated precision pulse capsulotomy vs manual capsulorhexis in white cataracts: reduction in procedural time and resource utilization.

PURPOSE: To compare the utility of precision pulse capsulotomy (PPC) with manual capsulorhexis for capsulotomy in white cataracts. SETTING: Hospital-based academic practice. DESIGN: Retrospective analysis of surgical case records and surgical videos from a single surgeon. METHODS: Cases involving intumescent and nonintumescent white cataracts were identified. Capsulotomy outcomes, surgical outcomes, procedural time, and resource utilization, as well as patient demographic and health data, were analyzed and subjected to statistical testing. RESULTS: 15 cases of white cataract (10 intumescent and 5 nonintumescent) performed using continuous curvilinear capsulorhexis (CCC) were compared with 20 cases (9 intumescent and 11 nonintumescent) performed using PPC. The cases covered a period of 14 months before and 30 months after surgeon adoption of PPC. There were no significant differences between the 2 groups in patient age, sex, ethnicity, ocular history, medical history, and medications. PPC resulted in complete capsulotomies without tags or tears and intracapsular intraocular lens implantation with 360-degree capsular overlap in all 20 cases. There was 1 CCC case resulting in the Argentinian flag sign. Compared with CCC, PPC white cataract cases also demonstrated significant advantages in capsulotomy time, reduced use of trypan blue and ophthalmic viscosurgical device, and less overall procedural time. CONCLUSIONS: PPC is a safe and highly effective method to create consistent capsulotomies in both intumescent and nonintumescent white cataracts. The use of PPC provides benefits of significant reductions in capsulotomy time, overall procedural time, and resource utilization, resulting in a streamlined treatment of these complex cataract surgery cases.

Endothelial Cell Loss Following Cataract Surgery Using Continuous Curvilinear Capsulorhexis or Precision Pulse Capsulotomy.

Purpose: To compare endothelial cell density (ECD), percentage of hexagonal cells (%Hex) and coefficient of variation (CV) in cell size following lens cataract surgery with phacoemulsification performed using Continuous Curvilinear Capsulorhexis (CCC) or Precision Pulse Capsulotomy (PPC). Patients and Methods: Sixty-seven subjects were randomly assigned to undergo lens cataract removal with the capsulotomy step performed using either CCC or PPC. Specular microscopy images were obtained pre-operatively, 1 month and 3 months after surgery. ECD, %Hex and CV were analyzed in a masked fashion by an independent reading center. Results: The mean percentage ECD loss at 1 month was 11.5% in the CCC group and 12.3% in the PPC group (P = 0.818; t-test). At 3 months, the mean percentage ECD loss was 11.7% in the CCC group and 12.4% in the PPC group (P = 0.815; t-test). The mean %Hex at 1 month was 54.3% in the CCC group and 54.7% in the PPC group (P = 0.695; t-test). At 3 months, the mean %Hex was 56.2% in the CCC group and 54.7% in the PPC group (P = 0.278; t-test). The CV at 1 month was 34.4% in the CCC group and 34.3% in the PPC group (P = 0.927; t-test). At 3 months, the CV was 32.7% in the CCC group and 33.4% in the PPC group (P = 0.864; t-test). Conclusion: No differences in ECD loss, %Hex and CV were observed between patients who received CCC or PPC. PPC use during cataract surgery does not result in any increased endothelial cell loss beyond that normally associated with this surgery.

Use of P1-P4 Purkinje reflections as a surrogate sign for intraoperative patient fixation.

Circumferential, even anterior capsular overlap maximizes intraocular lens stability and posterior capsular opacification mitigation and provides best long-term outcomes for the cataract patient. P1 and P4 Purkinje reflections at patient fixation may provide a reliable marker for capsulotomy centration. However, patient fixation may be hindered during surgery because of anesthesia or light sensitivity. In this study, we demonstrate that the relationship between the P1 and P4 Purkinje reflections previewed prior to surgery when the patient is fixating may be recreated intraoperatively if fixation becomes difficult. The final position of P1 and P4 relative to one another at fixation is invariant in a given patient, but there are variations among patients. Knowledge of the P1 and P4 relationship can be used as a surrogate sign of patient fixation to assist in capsulotomy centration during cataract surgery.

Precision pulse capsulotomy: performance metrics and utility in routine and complex cases.

PURPOSE: To evaluate precision pulse capsulotomy (PPC) performance. SETTING: University and private practice in the United States and South Korea. DESIGN: Multicenter retrospective analysis. METHODS: The surgical videos of 337 cataract surgeries with PPC capsulotomy performed by 4 surgeons at 4 centers were used to assess capsulotomy outcomes including completion rate, diameter, roundness (ovality), and quality of capsular overlap. RESULTS: PPC use resulted in 99.4% free-floating capsulotomies from 337 cases. Video image analysis in a subset (n = 52) yielded a mean capsulotomy diameter of 5.0 mm ± 0.16 mm SD (95% CI, 4.96-5.04 mm). Capsulotomies were round to slightly oval at the end of the case with a mean ovality of 3.0% ± 2.86% (95% CI, 2.22%-3.78%; 360 degrees capsular overlap was obtained in 98% of cases. The offset of the capsulotomy center with the intraocular lens (IOL) optic center was 197 µm ± 122 µm (SD) (95% CI, 148-246 µm). PPC was used successfully in traumatic cataracts with compromised anterior and posterior capsule, phacodonesis, intumescent cataract with constricted pupil, and zonular dialysis and in penetrating keratoplasty with open-sky extracapsular cataract extraction. CONCLUSIONS: Surgeons obtained good PPC capsulotomy outcomes in routine and challenging cases. Little variation was observed in achieving free-floating capsulotomies with approximately 5.0 mm diameter and complete capsular overlap. Variation was observed in the amount of offset between the capsulotomy center and the center of the IOL optic. PPC was useful in cases with multiple comorbidities that challenge capsulotomy performance.

Semaphorin 5A is a bifunctional axon guidance cue regulated by heparan and chondroitin sulfate proteoglycans.

The response of neuronal growth cones to axon guidance cues depends on the developmental context in which these cues are encountered. We show here that the transmembrane protein semaphorin 5A (Sema5A) is a bifunctional guidance cue exerting both attractive and inhibitory effects on developing axons of the fasciculus retroflexus, a diencephalon fiber tract associated with limbic function. The thrombospondin repeats of Sema5A physically interact with the glycosaminoglycan portion of both chondroitin sulfate proteoglycans (CSPGs) and heparan sulfate proteoglycans (HSPGs). CSPGs function as precisely localized extrinsic cues that convert Sema5A from an attractive to an inhibitory guidance cue. Therefore, glycosaminoglycan bound guidance cues provide a molecular mechanism for CSPG-mediated inhibition of axonal extension. Further, axonal HSPGs are required for Sema5A-mediated attraction, suggesting that HSPGs are components of functional Sema5A receptors. Thus, neuronal responses to Sema5A are proteoglycan dependent and interpreted according to the biological context in which this membrane bound guidance cue is presented.

FAK deficiency in cells contributing to the basal lamina results in cortical abnormalities resembling congenital muscular dystrophies.

Targeted deletion of focal adhesion kinase (fak) in the developing dorsal forebrain resulted in local disruptions of the cortical basement membrane located between the neuroepithelium and pia-meninges. At disruption sites, clusters of neurons invaded the marginal zone. Retraction of radial glial endfeet, midline fusion of brain hemispheres, and gliosis also occurred, similar to type II cobblestone lissencephaly as seen in congenital muscular dystrophy. Interestingly, targeted deletion of fak in neurons alone did not result in cortical ectopias, indicating that fak deletion from glia is required for neuronal mislocalization. Unexpectedly, fak deletion specifically from meningeal fibroblasts elicited similar cortical ectopias in vivo and altered laminin organization in vitro. These observations provide compelling evidence that FAK plays a key signaling role in cortical basement membrane assembly and/or remodeling. In addition, FAK is required within neurons during development because neuron-specific fak deletion alters dendritic morphology in the absence of lamination defects.

Microtechnology and nanotechnology in nerve repair.

OBJECTIVE: This review will describe the novel contributions to the field of nerve repair from the emerging disciplines of microtechnology and nanotechnology. METHOD: This broad review will cover the advances described in the literature of the medical and biological fields and the engineering and physical sciences. The authors have also included their own work in this field. DISCUSSION: Microtechnology and nanotechnology are providing two fundamentally different pathways for pursuing nerve repair: (1) microstructured scaffolds to promote regeneration and (2) direct repair by reconnecting axons. In the first instance, many of the traditional techniques for microfabrication of microelectronics have been applied to the development of implantable tissue scaffolds with precisely formed architectures. Combined with nanotechnological capabilities to control their surface chemistries, these tissue constructs have been designed to create a microenvironment within nerve tissue to optimally promote the outgrowth of neurites. With some initial successes in animal models, these next generation tissue scaffolds may provide a marked improvement over traditional nerve grafts in the ability to overcome nerve degenerative processes and to coax nerve regeneration leading to restoration of at least some nerve function. A second, completely different repair strategy aims to directly repair nerves at the microscale by acutely reconnecting severed or damaged axons immediately after injury and potentially forestalling the usual downstream degenerative processes. This strategy will take advantage of the traditional capabilities of microfabrication to create microelectromechanical systems that will serve as ultramicrosurgical tools that can operate at the micron scale and reliably manipulate individual axons without incurring damage. To bring about some restoration of a nerve's function, axon repair will have to be performed repetitively on a large scale and soon after injury. Development work is currently underway to bring about the feasibility of this technique. CONCLUSION: With the emergence of microtechnology and nanotechnology, new methods for repairing nerves are being explored and developed. There have been two fundamental benefits from the technologies of the ultrasmall scale: (1) enhancement of regeneration using new tissue scaffold materials and architecture; (2) direct repair of nerves at the scale of single neurons and axons.

Multifunctional biophotonic nanostructures inspired by the longtail glasswing butterfly for medical devices.

Numerous living organisms possess biophotonic nanostructures that provide colouration and other diverse functions for survival. While such structures have been actively studied and replicated in the laboratory, it remains unclear whether they can be used for biomedical applications. Here, we show a transparent photonic nanostructure inspired by the longtail glasswing butterfly (Chorinea faunus) and demonstrate its use in intraocular pressure (IOP) sensors in vivo. We exploit the phase separation between two immiscible polymers (poly(methyl methacrylate) and polystyrene) to form nanostructured features on top of a Si3N4 substrate. The membrane thus formed shows good angle-independent white-light transmission, strong hydrophilicity and anti-biofouling properties, which prevent adhesion of proteins, bacteria and eukaryotic cells. We then developed a microscale implantable IOP sensor using our photonic membrane as an optomechanical sensing element. Finally, we performed in vivo testing on New Zealand white rabbits, which showed that our device reduces the mean IOP measurement variation compared with conventional rebound tonometry without signs of inflammation.

EphB3: an endogenous mediator of adult axonal plasticity and regrowth after CNS injury.

Endogenous mechanisms underlying the remodeling of neuronal circuitry after mammalian CNS injury or disease remain primarily unknown. Here, we investigated axonal plasticity after optic nerve injury and found that macrophages recruited into the injury site and adult retinal ganglion cell (RGC) axons, which undergo injury-induced sprouting and terminal remodeling, were linked by their respective expression of a ligand and receptor pair active in axon guidance. Recruited macrophages specifically upregulated mRNA encoding the guidance molecule EphB3 and expressed EphB proteins capable of binding Ephrin B molecules in vivo and in vitro. Injured adult RGC axons in turn expressed EphrinB3, a known receptor for EphB3, and RGC axons bound recombinant EphB3 protein injected into the optic nerve. In vitro, EphB3 supported adult RGC axon outgrowth, and axons turned toward a source of this guidance molecule. In vivo, both reduction of EphB3 function in adult heterozygous animals and loss of function in homozygous animals greatly decreased RGC axon re-extension or sprouting after optic nerve injury. Comparisons of axon re-extension in EphB3 null and wild-type littermates showed that this loss of axonal plasticity was not attributable to a difference in intrinsic axon growth potential. Rather, the results indicated an essential role for local optic nerve-derived EphB3 in regulating adult RGC axon plasticity after optic nerve injury. Of note, the loss of EphB3 did not affect the ability of injured RGC axons to elaborate complex terminal branching, suggesting that additional EphB3-independent mechanisms governed adult axon branching triggered by CNS damage.

Upregulation of EphB2 and ephrin-B2 at the optic nerve head of DBA/2J glaucomatous mice coincides with axon loss.

PURPOSE: To identify genes with upregulated expression at the optic nerve head (ONH) that coincides with retinal ganglion cell (RGC) axon loss in glaucomatous DBA/2J mice. To further demonstrate that the proteins encoded by these genes bind to RGC axons and influence fundamental axon physiology. METHODS: In situ hybridization and cell-type-specific immunolabeling were performed on ONH sections from DBA/2J mice (3 to 11 months old) and C57Bl/6NCrl mice (10 months old). EphB2-Fc and ephrin-B2-Fc chimeric proteins were applied to adult RGC axons in vitro and in vivo at the ONH to demonstrate protein binding on axons. EphB2-Fc or control Fc protein was applied in a bath or locally to axons preloaded with the calcium indicator Fluo-4-AM, and changes in intra-axonal calcium were determined. RESULTS: EphB2 and ephrin-B2 were specifically upregulated at the ONH of DBA/2J mice starting at 9 months of age, but not in age-matched C57Bl/6NCrl mice or in DBA/2J animals that did not have axon loss. EphA4 was also present at the ONH, but no difference in expression was detected between unaffected and affected animals. EphB2 was expressed by F4/80(+), MOMA2(+), ED1(-) macrophage-like cells, ephrin-B2 was expressed by Iba-1(+) microglia and GFAP(+) astrocytes, whereas EphA4 was expressed by GFAP(+) astrocytes. EphB2-Fc and ephrin-B2-Fc protein bound to RGC axons in culture and to ONH RGC axons in vivo. Adult RGC axons in vitro elevated intra-axonal calcium in response to EphB2-Fc but not to control Fc protein. CONCLUSIONS: The expression of EphB2 and ephrin-B2 is upregulated at the ONH of glaucomatous DBA/2J mice coinciding with RGC axon loss. The direct binding of EphB2 and ephrin-B2 on adult RGC axons at the ONH and the ability of EphB2 to elevate intra-axonal calcium indicate that these proteins may affect RGC axon physiology in the setting of glaucoma and thus affect the development or progression of the disease.

Microtechnology in medicine: the emergence of surgical microdevices.

With the emergence of technologies to fabricate and mass-produce microscale tools and micromachines, microsurgery stands to potentially benefit through the development of a fundamentally new class of instruments. These new instruments may provide the surgeon with access to the smallest reaches of the body and perform operations that are currently not possible with manually operated tools. These new devices can be variably constructed and configured based on a wide range of design possibilities and can be built to serve many different fundamental surgical functions requiring the manipulation and handling of small tissues and structures, including grasping, cutting, and monitoring. With these functionalities also comes a high degree of integration, allowing tools and space to be used efficiently. Adapted from the techniques of the microelectronics industry, the fabrication methods and materials produce structures that are mechanically strong and easy to reproduce on a large scale. Well-developed design and physical modeling tools mean that the process of instrument development and validation can be streamlined. Along with these new instruments comes the need to provide automated interfaces to effectively translate human operator intentions into the appropriate actuation and motion of these devices. These interfaces must include the capability to scale down human motions to the range of microns. Most likely, the operation of these new microsurgical devices will resemble the control schemes developed for robotic surgery. The control schemes will provide accurate motions while minimizing the chances of damaging tools or unnecessarily injuring tissues. Naturally, these new tools and surgical schemes will require a transition from the conventional paradigm. However, with new surgical capabilities that may allow direct intervention into the inner workings of a cell, MEMS and nanotechnology-based tools may become a crucial part of the arsenal for the next generation of surgeons. Invariably, future developments of this new class of instruments will depend in large part on needs identified by the surgeon and an understanding of the enabling properties of microtechnology and nanotechnology. Thus, recognition of the vast potentials of this new technology among clinicians will greatly help to accelerate the development and integration of new microdevices and novel procedures that address disease and injury with unprecedented precision.

A microscale optical implant for continuous in vivo monitoring of intraocular pressure.

Intraocular pressure (IOP) is a key clinical parameter in glaucoma management. However, despite the potential utility of daily measurements of IOP in the context of disease management, the necessary tools are currently lacking, and IOP is typically measured only a few times a year. Here we report on a microscale implantable sensor that could provide convenient, accurate, on-demand IOP monitoring in the home environment. When excited by broadband near-infrared (NIR) light from a tungsten bulb, the sensor's optical cavity reflects a pressure-dependent resonance signature that can be converted to IOP. NIR light is minimally absorbed by tissue and is not perceived visually. The sensor's nanodot-enhanced cavity allows for a 3-5 cm readout distance with an average accuracy of 0.29 mm Hg over the range of 0-40 mm Hg. Sensors were mounted onto intraocular lenses or silicone haptics and secured inside the anterior chamber in New Zealand white rabbits. Implanted sensors provided continuous in vivo tracking of short-term transient IOP elevations and provided continuous measurements of IOP for up to 4.5 months.

Biocompatible Multifunctional Black-Silicon for Implantable Intraocular Sensor.

Multifunctional black-silicon (b-Si) integrated on the surface of an implantable intraocular pressure sensor significantly improves sensor performance and reliability in six-month in vivo studies. The antireflective properties of b-Si triples the signal-to-noise ratio and increases the optical readout distance to a clinically viable 12 cm. Tissue growth and inflammation response on the sensor is suppressed demonstrating desirable anti-biofouling properties.

Isolation of neuronal substructures and precise neural microdissection using a nanocutting device.

We describe a set of microfabricated nanocutting devices with a cutting edge of less than 20 nm radius of curvature that enables high precision microdissection and subcellular isolation of neuronal structures. With these devices, it is possible to isolate functional substructures from neurons in culture such as segments of axons and dendrites, dendritic spines and Nodes of Ranvier. By fine-tuning the mechanical compliance of these devices, they can also act as alternatives to costly laser capture microdissection workstations for harvesting specific neuronal populations from tissue sections for analysis. The small size of the device (1 mm2x100 microm) allows convenient insertion into researcher specific experimental set-ups. Its ease of use and possibility for batch fabrication makes this a highly effective and versatile tool for tissue microdissection and the microanalysis of neuronal function.

L1/Laminin modulation of growth cone response to EphB triggers growth pauses and regulates the microtubule destabilizing protein SCG10.

During development, EphB proteins serve as axon guidance molecules for retinal ganglion cell axon pathfinding toward the optic nerve head and in midbrain targets. To better understand the mechanisms by which EphB proteins influence retinal growth cone behavior, we investigated how axon responses to EphB were modulated by laminin and L1, two guidance molecules that retinal axons encounter during in vivo pathfinding. Unlike EphB stimulation in the presence of laminin, which triggers typical growth cone collapse, growth cones co-stimulated by L1 did not respond to EphB. Moreover, EphB exposure in the presence of both laminin and L1 resulted in a novel growth cone inhibition manifested as a pause in axon elongation with maintenance of normal growth cone morphology and filopodial activity. Pauses were not associated with loss of growth cone actin but were accompanied by a redistribution of the microtubule cytoskeleton with increased numbers of microtubules extending into filopodia and to the peripheral edge of the growth cone. This phenomenon was accompanied by reduced levels of the growth cone microtubule destabilizing protein SCG10. Antibody blockade of SCG10 function in growth cones resulted in both changes in microtubule distribution and pause responses mirroring those elicited by EphB in the presence of laminin and L1. These results demonstrate that retinal growth cone responsiveness to EphB is regulated by co-impinging signals from other axon guidance molecules. Furthermore, the results are consistent with EphB-mediated axon guidance mechanisms that involve the SCG10-mediated regulation of the growth cone microtubule cytoskeleton.

An oligodendrocyte lineage-specific semaphorin, Sema5A, inhibits axon growth by retinal ganglion cells.

In the mammalian CNS, glial cells repel axons during development and inhibit axon regeneration after injury. It is unknown whether the same repulsive axon guidance molecules expressed by glia and their precursors during development also play a role in inhibiting regeneration in the injured CNS. Here we investigate whether optic nerve glial cells express semaphorin family members and, if so, whether these semaphorins inhibit axon growth by retinal ganglion cells (RGCs). We show that each optic nerve glial cell type, astrocytes, oligodendrocytes, and their precursor cells, expressed a distinct complement of semaphorins. One of these, sema5A, was expressed only by purified oligodendrocytes and their precursors, but not by astrocytes, and was present in both normal and axotomized optic nerve but not in peripheral nerves. Sema5A induced collapse of RGC growth cones and inhibited RGC axon growth when presented as a substrate in vitro. To determine whether sema5A might contribute to inhibition of axon growth after injury, we studied the ability of RGCs to extend axons when cultured on postnatal day (P) 4, P8, and adult optic nerve explants and found that axon growth was strongly inhibited. Blocking sema5A using a neutralizing antibody significantly increased RGC axon growth on these optic nerve explants. These data support the hypothesis that sema5A expression by oligodendrocyte lineage cells contributes to the glial cues that inhibit CNS regeneration.

Regulation of axon regeneration by the RNA repair and splicing pathway.

Mechanisms governing a neuron's regenerative ability are important but not well understood. We identify Rtca (RNA 3'-terminal phosphate cyclase) as an inhibitor of axon regeneration. Removal of Rtca cell-autonomously enhanced axon regrowth in the Drosophila CNS, whereas its overexpression reduced axon regeneration in the periphery. Rtca along with the RNA ligase Rtcb and its catalyst Archease operate in the RNA repair and splicing pathway important for stress-induced mRNA splicing, including that of Xbp1, a cellular stress sensor. Drosophila Rtca and Archease had opposing effects on Xbp1 splicing, and deficiency of Archease or Xbp1 impeded axon regeneration in Drosophila. Moreover, overexpressing mammalian Rtca in cultured rodent neurons reduced axonal complexity in vitro, whereas reducing its function promoted retinal ganglion cell axon regeneration after optic nerve crush in mice. Our study thus links axon regeneration to cellular stress and RNA metabolism, revealing new potential therapeutic targets for treating nervous system trauma.