Proximity-based activation of AURORA A by MPS1 potentiates error correction.

Faithfull cell division relies on mitotic chromosomes becoming bioriented with each pair of sister kinetochores bound to microtubules oriented toward opposing spindle poles. Erroneous kinetochore-microtubule attachments often form during early mitosis, but are destabilized through the phosphorylation of outer kinetochore proteins by centromeric AURORA B kinase (ABK) and centrosomal AURORA A kinase (AAK), thus allowing for re-establishment of attachments until biorientation is achieved. MPS1-mediated phosphorylation of NDC80 has also been shown to directly weaken the kinetochore-microtubule interface in yeast. In human cells, MPS1 has been proposed to transiently accumulate at end-on attached kinetochores and phosphorylate SKA3 to promote microtubule release. Whether MPS1 directly targets NDC80 and/or promotes the activity of AURORA kinases in metazoans remains unclear. Here, we report a novel mechanism involving communication between kinetochores and centrosomes, wherein MPS1 acts upstream of AAK to promote error correction. MPS1 on pole-proximal kinetochores phosphorylates the C-lobe of AAK thereby increasing its activation at centrosomes. This proximity-based activation ensures the establishment of a robust AAK activity gradient that locally destabilizes mal-oriented kinetochores near spindle poles. Accordingly, MPS1 depletion from Drosophila cells causes severe chromosome misalignment and erroneous kinetochore-microtubule attachments, which can be rescued by tethering either MPS1 or constitutively active AAK mutants to centrosomes. Proximity-based activation of AAK by MPS1 also occurs in human cells to promote AAK-mediated phosphorylation of the NDC80 N-terminal tail. These findings uncover an MPS1-AAK cross-talk that is required for efficient error correction, showcasing the ability of kinetochores to modulate centrosome outputs to ensure proper chromosome segregation.

Deep Learning Segmentation of the Nucleus Basalis of Meynert on 3T MRI.

BACKGROUND AND PURPOSE: The nucleus basalis of Meynert is a key subcortical structure that is important in arousal and cognition and has been explored as a deep brain stimulation target but is difficult to study due to its small size, variability among patients, and lack of contrast on 3T MR imaging. Thus, our goal was to establish and evaluate a deep learning network for automatic, accurate, and patient-specific segmentations with 3T MR imaging. MATERIALS AND METHODS: Patient-specific segmentations can be produced manually; however, the nucleus basalis of Meynert is difficult to accurately segment on 3T MR imaging, with 7T being preferred. Thus, paired 3T and 7T MR imaging data sets of 21 healthy subjects were obtained. A test data set of 6 subjects was completely withheld. The nucleus was expertly segmented on 7T, providing accurate labels for the paired 3T MR imaging. An external data set of 14 patients with temporal lobe epilepsy was used to test the model on brains with neurologic disorders. A 3D-Unet convolutional neural network was constructed, and a 5-fold cross-validation was performed. RESULTS: The novel segmentation model demonstrated significantly improved Dice coefficients over the standard probabilistic atlas for both healthy subjects (mean, 0.68 [SD, 0.10] versus 0.45 [SD, 0.11], P = .002, t test) and patients (0.64 [SD, 0.10] versus 0.37 [SD, 0.22], P < .001). Additionally, the model demonstrated significantly decreased centroid distance in patients (1.18 [SD, 0.43] mm, 3.09 [SD, 2.56] mm, P = .007). CONCLUSIONS: We developed the first model, to our knowledge, for automatic and accurate patient-specific segmentation of the nucleus basalis of Meynert. This model may enable further study into the nucleus, impacting new treatments such as deep brain stimulation.

Evolution of territoriality in Hylinae treefrogs: Ecological and morphological correlates and lineage diversification.

Given the diverse nature of traits involved in territorial defence, they may respond to different selective pressures and then exhibit distinct patterns of evolution. These selective pressures also may cause territorial behaviour to be associated with environmental and morphological variables. Such associations, however, have mostly been studied at the intraspecific level, being phylogenetic analyses of territoriality in a broad taxonomic framework rare in the literature. We used the anuran subfamily Hylinae to test (1) whether two territorial-behaviour traits with different levels of aggression-territorial call and physical combat-are evolutionarily more labile than a morphological trait used in physical combat-the spine-shaped prepollex; (2) whether reproduction in lentic waters and phytotelmata, as well as resource scarcity, might favour the occurrence of territoriality; (3) if physical combat is more important than territorial call for the evolution of body size and sexual size dimorphism and (4) the relationships between territorial-behaviour traits and lineage diversification. We mainly used the literature to build two datasets with different levels of certainty. Territorial-behaviour traits exhibited intermediate levels of phylogenetic signal in Hylinae, whereas the phylogenetic signal for the presence of the spine-shaped prepollex was strong. We found support for the hypothesis that reproduction in lentic water favours the occurrence of territorial behaviour, because the expression of territorial-behaviour traits was more associated with reproduction in lentic than in lotic waters. Territorial-behaviour traits were not correlated with annual precipitation nor with habitat complexity. Body size and sexual size dimorphism were not correlated with the presence of territorial call nor with physical combat. We identified negative correlations between diversification rates and physical combat. Relationships of territorial call and physical combat with diversification rates suggest that these territorial behaviours influence evolutionary processes in different ways.

Disparity in the use of Alzheimer's disease treatment in Southern Brazil.

Alzheimer's disease (AD) treatment is freely available in the Brazilian public health system. However, the prescription pattern and its associated factors have been poorly studied in our country. We reviewed all granted requests for AD treatment in the public health system in October 2021 in the Rio Grande do Sul (RS) state, Southern Brazil. We performed a spatial autocorrelation analysis with the population-adjusted patients receiving any AD medication as the outcome and correlated it with several socioeconomic variables. 2382 patients with AD were being treated during the period analyzed. The distribution of the outcome variable was not random (Moran's I 0.17562, P <.0001), with the most developed regions having a higher number of patients/100,000 receiving any AD medication. We show that although AD medications are available through the public health system, there is a clear disparity between regions of RS state. Factors related to socioeconomic development partly explain this finding.

Long-Term Neurophysiological Adaptations to Strength Training: A Systematic Review With Cross-Sectional Studies.

ABSTRACT: Santos, PDG, Vaz, JR, Correia, J, Neto, T, and Pezarat-Correia, P. Long-term neurophysiological adaptations to strength training: a systematic review with cross-sectional studies. J Strength Cond Res 37(10): 2091-2105, 2023-Neuromuscular adaptations to strength training are an extensively studied topic in sports sciences. However, there is scarce information about how neural mechanisms during force production differ between trained and untrained individuals. The purpose of this systematic review is to better understand the differences between highly trained and untrained individuals to establish the long-term neural adaptations to strength training. Three databases were used for the article search (PubMed, Web of Science, and Scopus). Studies were included if they compared groups of resistance-trained with untrained people, aged 18-40 year, and acquired electromyography (EMG) signals during strength tasks. Twenty articles met the eligibility criteria. Generally, strength-trained individuals produced greater maximal voluntary activation, while reducing muscle activity in submaximal tasks, which may affect the acute response to strength training. These individuals also presented lower co-contraction of the antagonist muscles, although it depends on the specific training background. Global intermuscular coordination may be another important mechanism of adaptation in response to long-term strength training; however, further research is necessary to understand how it develops over time. Although these results should be carefully interpreted because of the great disparity of analyzed variables and methods of EMG processing, chronic neural adaptations seem to be decisive to greater force production. It is crucial to know the timings at which these adaptations stagnate and need to be stimulated with advanced training methods. Thus, training programs should be adapted to training status because the same stimulus in different training stages will lead to different responses.

Gamma knife capsulotomy for intractable OCD: Neuroimage analysis of lesion size, location, and clinical response.

Obsessive-compulsive disorder (OCD) affects 2-3% of the population. One-third of patients are poorly responsive to conventional therapies, and for a subgroup, gamma knife capsulotomy (GKC) is an option. We examined lesion characteristics in patients previously treated with GKC through well-established programs in Providence, RI (Butler Hospital/Rhode Island Hospital/Alpert Medical School of Brown University) and São Paulo, Brazil (University of São Paolo). Lesions were traced on T1 images from 26 patients who had received GKC targeting the ventral half of the anterior limb of the internal capsule (ALIC), and the masks were transformed into MNI space. Voxel-wise lesion-symptom mapping was performed to assess the influence of lesion location on Y-BOCS ratings. General linear models were built to compare the relationship between lesion size/location along different axes of the ALIC and above or below-average change in Y-BOCS ratings. Sixty-nine percent of this sample were full responders (≥35% improvement in OCD). Lesion occurrence anywhere within the targeted region was associated with clinical improvement, but modeling results demonstrated that lesions occurring posteriorly (closer to the anterior commissure) and dorsally (closer to the mid-ALIC) were associated with the greatest Y-BOCS reduction. No association was found between Y-BOCS reduction and overall lesion volume. GKC remains an effective treatment for refractory OCD. Our data suggest that continuing to target the bottom half of the ALIC in the coronal plane is likely to provide the dorsal-ventral height required to achieve optimal outcomes, as it will cover the white matter pathways relevant to change. Further analysis of individual variability will be essential for improving targeting and clinical outcomes, and potentially further reducing the lesion size necessary for beneficial outcomes.

Relationships among total mixed ration nutritional components and reproductive performance in high-producing dairy herds.

The main objective of the present study was to determine whether composition of total mixed ration influences reproductive performance in high-producing commercial dairy farms. Dairy producers and nutritional consultants from 48 dairy farms located in Wisconsin agreed to provide reproductive data and dietary information on high milk production pens during the main breeding period for the previous 12 mo. Dietary components (percentage of dry matter) were crude protein (CP), rumen degradable (RDP) and undegradable (RUP) protein, neutral detergent fiber (NDF), nonfiber carbohydrates (NFC), starch, and fat. Reproductive data were service rate (SR), overall pregnancy per artificial insemination (P/AI) and P/AI at the first service, 21-d pregnancy rate (PR), days open, and percentage of cows pregnant by 150 d in milk (PREG150). Participating herds had lactating Holstein cows (range = 143 to 2,717) housed in freestall facilities. Statistical analyses were performed with CORR and GLIMMIX of SAS (SAS Institute Inc.). Daily average milk production of herds was 38.9 ± 0.60 kg/d (30.0 to 50.4 kg/d). Overall SR was 58.5% (39-73) and P/AI was 36.1% (22-49). Overall 21-d PR was 20.3% (10-42%). Correlation between SR and PR was 0.59, whereas correlation of overall P/AI and P/AI at first service with PR were both 0.72. Similarly, for PREG150, correlation with overall P/AI (0.63) and P/AI at first service (0.66) were greater than with SR (0.48). There was large variation in diet composition, with CP varying from 16.0 to 18.7%, NDF from 24.9 to 35.1%, NFC from 31.7 to 46.6%, starch from 20.1 to 30.8%, and fat from 3.1 to 6.7%. Overall, there were no detectable associations of CP, RDP, and RUP with reproductive measures. The strongest relationship was a decrease in reproductive performance with increasing dietary NFC including overall P/AI (-0.48), P/AI at first service (-0.51), and PREG150 (-0.33). Starch also had a negative relationship with P/AI at first service (-0.35). Conversely, greater NDF was positively associated with P/AI at first service (0.34). Fat content was also positively associated with P/AI at first service (0.34). When NFC was divided in tertiles (<40, 40 to 42.2, and >42.2% NFC), the highest tertile had lower overall P/AI (39 vs. 36 vs. 31%), P/AI at first service (43 vs. 40 vs. 33%), and PREG150 (54 vs. 53 vs. 47%). In conclusion, farms with greater dietary NFC may have compromised reproductive performance. Correspondingly, herds with greater NDF content may achieve high milk production with potentially positive associations with reproduction. Other relationships of dietary components on reproduction were not as obvious in this herd-level analysis.

Numerical Design of a Thread-Optimized Gripping System for Lap Joint Testing in a Split Hopkinson Apparatus.

Currently, few experimental methods exist that enable the mechanical characterization of adhesives under high strain rates. One such method is the Split Hopkinson Bar (SHB) test. The mechanical characterization of adhesives is performed using different specimen configurations, such as Single Lap Joint (SLJ) specimens. A gripping system, attached to the bars through threading, was conceived to enable the testing of SLJs. An optimization study for selecting the best thread was performed, analyzing the thread type, the nominal diameter, and the thread pitch. Afterwards, the gripping system geometry was numerically evaluated. The optimal threaded connection for the specimen consists of a trapezoidal thread with a 14 mm diameter and a 2 mm thread pitch. To validate the gripping system, the load-displacement (P-δ) curve of an SLJ, which was simulated as if it were tested on the SHB apparatus, was compared with an analogous curve from a validated drop-weight test numerical model.

Effect of Time-Restricted Eating and Resistance Training on High-Speed Strength and Body Composition.

This study examined the effects of four weeks of resistance training combined with time-restricted eating (TRE) vs. habitual diet on fat and fat-free mass as well as maximum and explosive force production in healthy, trained participants (18 males, aged 23.7 ± 2.6 years). The order of dieting was randomized and counterbalanced, and the participants served as their own controls. TRE involved an 8-h eating window and non-TRE involved a habitual meal pattern. Participants completed performance strength tests and body composition scans at baseline and post-intervention. The participants followed a structured training routine during each dietary intervention (four sets of maximum repetitions at 85% 1RM in five dynamic exercises, three times/week). Both interventions elicited deceases in fat mass (p < 0.05) but not in fat-free mass. After training (controlling for baseline values as covariates), non-TRE was compatible with better lower body jump performance than TRE (p < 0.05). Conversely, training with TRE elicited higher values in terms of peak force and dynamic strength index at the level of the upper body (p < 0.05). Thus, it can be concluded that there were no differences in fat mass and fat-free mass changes between interventions in already trained young males. Additionally, while the combination of TRE and resistance training might be beneficial for individuals focusing on developing high-speed strength performance at the upper body level, this is not applicable to those focusing on training the lower body.

Exploring the response of a key Mediterranean gorgonian to heat stress across biological and spatial scales.

Understanding the factors and processes that shape intra-specific sensitivity to heat stress is fundamental to better predicting the vulnerability of benthic species to climate change. Here, we investigate the response of a habitat-forming Mediterranean octocoral, the red gorgonian Paramuricea clavata (Risso, 1826) to thermal stress at multiple biological and geographical scales. Samples from eleven P. clavata populations inhabiting four localities separated by hundreds to more than 1500 km of coast and with contrasting thermal histories were exposed to a critical temperature threshold (25 °C) in a common garden experiment in aquaria. Ten of the 11 populations lacked thermotolerance to the experimental conditions provided (25 days at 25 °C), with 100% or almost 100% colony mortality by the end of the experiment. Furthermore, we found no significant association between local average thermal regimes nor recent thermal history (i.e., local water temperatures in the 3 months prior to the experiment) and population thermotolerance. Overall, our results suggest that local adaptation and/or acclimation to warmer conditions have a limited role in the response of P. clavata to thermal stress. The study also confirms the sensitivity of this species to warm temperatures across its distributional range and questions its adaptive capacity under ocean warming conditions. However, important inter-individual variation in thermotolerance was found within populations, particularly those exposed to the most severe prior marine heatwaves. These observations suggest that P. clavata could harbor adaptive potential to future warming acting on standing genetic variation (i.e., divergent selection) and/or environmentally-induced phenotypic variation (i.e., intra- and/or intergenerational plasticity).

Experimental and Numerical Study of Thermal Residual Stresses on Multimaterial Adherends in Single-Lap Joints.

The presence of residual stresses in composite materials can significantly affect material performance, especially when integrated in bonded joints. These stresses, often generated during the cure process, can cause cracking and distortion of the material, and are caused by differences in the coefficients of thermal expansion or cure shrinkage. In the current research, multimaterial adherends combining carbon-fibre-reinforced polymer (CFRP) and aluminium in a single-lap joint (SLJ) configuration are analysed, allowing us to understand the effect of the thermal residual stresses, developed during the curing process, in the overall performance of the joints. A numerical model resorting to a finite element analysis (FEA) is developed to assess and predict the behaviour of the joints. The use of FML (fibre metal laminates) was found to significantly improve the strength of the joints, as well as the failure mode. The proposed geometry performed similarly to the comparable FML geometry, in addition to a decrease in the joint weight.

Enhanced thermal and photo-stability of a para-substituted dicumyl ketone intercalated in a layered double hydroxide.

A ketodiacid, 4,4'-dicarboxylate-dicumyl ketone (3), has been intercalated into a Zn, Al layered double hydroxide (LDH) by a coprecipitation synthesis strategy. The structure and chemical composition of the resultant hybrid material (LDH-KDA3) were characterized by powder X-ray diffraction (PXRD), FT-IR, FT-Raman and solid-state 13C{1H} NMR spectroscopies, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), thermogravimetric analysis (TGA), and elemental analysis (CHN). PXRD showed that the dicarboxylate guest molecules assembled into a monolayer to give a basal spacing of 18.0 Å. TGA revealed that the organic guest starts to decompose at a significantly higher temperature (ca. 330°C) than that determined for the free ketodiacid (ca. 230°C). Photochemical experiments were performed to probe the photoreactivity of the ketoacid in the crystalline state, in solution, and as a guest embedded within the photochemically-inert LDH host. Irradiation of the bulk crystalline ketoacid results in photodecarbonylation and the exclusive formation of the radical-radical combination product. Solution studies employing the standard myoglobin (Mb) assay for quantification of released CO showed that the ketoacid behaved as a photoactivatable CO-releasing molecule for transfer of CO to heme proteins, although the photoreactivity was low. No photoinduced release of CO was found for the LDH system, indicating that molecular confinement enhanced the photo-stability of the hexasubstituted ketone. To better understand the behavior of 3 under irradiation, a more comprehensive study, involving excitation of this compound in DMSO-d6 followed by 1H NMR, UV-Vis and fluorescence spectroscopy, was undertaken and further rationalized with the help of time-dependent density functional theory (TDDFT) electronic quantum calculations. The photophysical study showed the formation of a less emissive compound (or compounds). New signals in the 1H NMR spectra were attributed to photoproducts obtained via Norrish type I α-cleavage decarbonylation and Norrish type II (followed by CH3 migration) pathways. TDDFT calculations predicted that the formation of a keto-enol system (via a CH3 migration step in the type II pathway) was highly favorable and consistent with the observed spectral data.

Imaging Aspects of the Hippocampus.

The hippocampus is one of the most sophisticated structures in the brain, owing to its complex anatomy, intriguing functions, relationship with other structures, and relevant associated symptoms. Despite being a structure analyzed for centuries, its anatomy and physiology in the human body are still being extensively studied, as well as associated pathologic conditions and potential biomarkers. It can be affected by a broad group of diseases that can be classified as congenital, degenerative, infectious or inflammatory, neoplastic, vascular, or toxic-metabolic disease. The authors present the anatomy and close structures, function, and development of the hippocampus, as well as an original algorithm for imaging diagnosis. The algorithm includes pathologic conditions that typically affect the hippocampus and groups them into nodular (space occupying) and nonnodular pathologic conditions, serving as a guide to narrow the differential diagnosis. MRI is the imaging modality of choice for evaluation of the hippocampus, and CT and nuclear medicine also improve the analysis. The MRI differential diagnosis depends on anatomic recognition and careful characterization of associated imaging findings such as volumetric changes, diffusion restriction, cystic appearance, hyperintensity at T1-weighted imaging, enhancement, or calcification, which play a central role in diagnosis along with clinical findings. Some pathologic conditions arising from surrounding structures such as the amygdala are also important to recognize. Pathologic conditions of the hippocampus can be a challenge to diagnose because they usually manifest as similar clinical syndromes, so the imaging findings play a potential role in guiding the final diagnosis. Online supplemental material is available for this article. ©RSNA, 2022.

Intermuscular Coordination in the Power Clean Exercise: Comparison between Olympic Weightlifters and Untrained Individuals-A Preliminary Study.

Muscle coordination in human movement has been assessed through muscle synergy analysis. In sports science, this procedure has been mainly applied to the comparison between highly trained and unexperienced participants. However, the lack of knowledge regarding strength training exercises led us to study the differences in neural strategies to perform the power clean between weightlifters and untrained individuals. Synergies were extracted from electromyograms of 16 muscles of ten unexperienced participants and seven weightlifters. To evaluate differences, we determined the pairwise correlations for the synergy components and electromyographic profiles. While the shape of activation patterns presented strong correlations across participants of each group, the weightings of each muscle were more variable. The three extracted synergies were shifted in time with the unexperienced group anticipating synergy #1 (-2.46 ± 18.7%; p < 0.001) and #2 (-4.60 ± 5.71%; p < 0.001) and delaying synergy #3 (1.86 ± 17.39%; p = 0.01). Moreover, muscle vectors presented more inter-group variability, changing the composition of synergy #1 and #3. These results may indicate an adaptation in intermuscular coordination with training, and athletes in an initial phase of training should attempt to delay the hip extension (synergy #1), as well as the upper-limb flexion (synergy #2).

Cholesterol synthesis inhibition promotes axonal regeneration in the injured central nervous system.

Neuronal regeneration in the injured central nervous system is hampered by multiple extracellular proteins. These proteins exert their inhibitory action through interactions with receptors that are located in cholesterol rich compartments of the membrane termed lipid rafts. Here we show that cholesterol-synthesis inhibition prevents the association of the Neogenin receptor with lipid rafts. Furthermore, we show that cholesterol-synthesis inhibition enhances axonal growth both on inhibitory -myelin and -RGMa substrates. Following optic nerve injury, lowering cholesterol synthesis with both drugs and siRNA-strategies allows for robust axonal regeneration and promotes neuronal survival. Cholesterol inhibition also enhanced photoreceptor survival in a model of Retinitis Pigmentosa. Our data reveal that Lovastatin leads to several opposing effects on regenerating axons: cholesterol synthesis inhibition promotes regeneration whereas altered prenylation impairs regeneration. We also show that the lactone prodrug form of lovastatin has differing effects on regeneration when compared to the ring-open hydroxy-acid form. Thus the association of cell surface receptors with lipid rafts contributes to axonal regeneration inhibition, and blocking cholesterol synthesis provides a potential therapeutic approach to promote neuronal regeneration and survival in the diseased Central Nervous System. SIGNIFICANCE STATEMENT: Statins have been intensively used to treat high levels of cholesterol in humans. However, the effect of cholesterol inhibition in both the healthy and the diseased brain remains controversial. In particular, it is unclear whether cholesterol inhibition with statins can promote regeneration and survival following injuries. Here we show that late stage cholesterol inhibition promotes robust axonal regeneration following optic nerve injury. We identified distinct mechanisms of action for activated vs non-activated Lovastatin that may account for discrepancies found in the literature. We show that late stage cholesterol synthesis inhibition alters Neogenin association with lipid rafts, thereby i) neutralizing the inhibitory function of its ligand and ii) offering a novel opportunity to promote CNS regeneration and survival following injuries.

The Borel map in locally integrable structures.

Given a locally integrable structure V over a smooth manifold Ω and given p ∈ Ω we define the Borel map of V at p as the map which assigns to the germ of a smooth solution of V at p its formal Taylor power series at p. In this work we continue the study initiated in Barostichi et al. (Math. Nachr. 286(14-15):1439-1451, 2013), Della Sala and Lamel (Int J Math 24(11):1350091, 2013) and present new results regarding the Borel map. We prove a general necessary condition for the surjectivity of the Borel map to hold and also, after developing some new devices, we study some classes of CR structures for which its surjectivity is valid. In the final sections we show how the Borel map can be applied to the study of the algebra of germs of solutions of V at p.

Muscle Synergies Reliability in the Power Clean Exercise.

Muscle synergy extraction has been utilized to investigate muscle coordination in human movement, namely in sports. The reliability of the method has been proposed, although it has not been assessed previously during a complex sportive task. Therefore, the aim of the study was to evaluate intra- and inter-day reliability of a strength training complex task, the power clean, assessing participants' variability in the task across sets and days. Twelve unexperienced participants performed four sets of power cleans in two test days after strength tests, and muscle synergies were extracted from electromyography (EMG) data of 16 muscles. Three muscle synergies accounted for almost 90% of variance accounted for (VAF) across sets and days. Intra-day VAF, muscle synergy vectors, synergy activation coefficients and individual EMG profiles showed high similarity values. Inter-day muscle synergy vectors had moderate similarity, while the variables regarding temporal activation were still strongly related. The present findings revealed that the muscle synergies extracted during the power clean remained stable across sets and days in unexperienced participants. Thus, the mathematical procedure for the extraction of muscle synergies through nonnegative matrix factorization (NMF) may be considered a reliable method to study muscle coordination adaptations from muscle strength programs.

Stabilization of primary cilia reduces abortive cell cycle re-entry to protect injured adult CNS neurons from apoptosis.

Abortive cell cycle (ACC) re-entry of apoptotic neurons is a recently characterized phenomenon that occurs after central nervous system (CNS) injury or over the course of CNS disease. Consequently, inhibiting cell cycle progression is neuroprotective in numerous CNS pathology models. Primary cilia are ubiquitous, centriole-based cellular organelles that prevent cell cycling, but their ability to modulate abortive cell cycle has not been described. Here, we show that neuronal cilia are ablated in-vitro and in-vivo following injury by hypoxia or optic nerve transection (ONT), respectively. Furthermore, forced cilia resorption sensitized neurons to these injuries and enhanced cell death. In contrast, pharmacological inhibition or shRNA knockdown of the proteins that disassemble the cilia increased neuron survival and decreased the phosphorylation of retinoblastoma (Rb), a master switch for cell cycle re-entry. Our findings show that the stabilization of neuronal primary cilia inhibits, at least transiently, apoptotic cell cycling, which has implications for future therapeutic strategies that halt or slow the progression of neurodegenerative diseases and acute CNS injuries.

MMP Inhibition Preserves Integrin Ligation and FAK Activation to Induce Survival and Regeneration in RGCs Following Optic Nerve Damage.

Purpose: Integrin adherence to the extracellular matrix (ECM) is essential for retinal ganglion cell (RGC) survival: damage causes production and release of ECM degrading matrix metalloproteinases (MMPs) that disrupt integrin ligation, leading to RGC death. The interplay of MMPs, integrins, and focal adhesion kinase (FAK) was studied in RGCs after optic nerve injury. Methods: Optic nerve transection and optic nerve crush were used to study RGC survival and regeneration, respectively. Treatments were administered intravitreally or into the cut end of the optic nerve. RGC survival was assessed by fluorescence or confocal microscopy; cell counting, peptide levels, and localization were assessed by Western blot and immunohistochemistry. Results: MMP-9 was most strongly increased and localized to RGCs after injury. Pan-MMP, MMP-2/-9, and MMP-3 inhibition all significantly enhanced RGC survival and increased RGC axon regeneration. FAK activation was decreased at 4 days postaxotomy, when apoptosis begins. FAK inhibition reduced RGC survival and abrogated the neuroprotective effects of MMP inhibition, whereas FAK activation increased RGC survival despite MMP activation. Integrin ligation with CD29 antibody or glycine-arginine-glycine-aspatate-serine (GRGDS) peptide increased RGC survival after axotomy. Conclusions: ECM-integrin ligation promotes RGC survival and axon regeneration via FAK activation.