Alternative splicing controls teneurin-3 compact dimer formation for neuronal recognition.

Neuronal network formation is facilitated by recognition between synaptic cell adhesion molecules at the cell surface. Alternative splicing of cell adhesion molecules provides additional specificity in forming neuronal connections. For the teneurin family of cell adhesion molecules, alternative splicing of the EGF-repeats and NHL domain controls synaptic protein-protein interactions. Here we present cryo-EM structures of the compact dimeric ectodomain of two teneurin-3 isoforms that harbour the splice insert in the EGF-repeats. This dimer is stabilised by an EGF8-ABD contact between subunits. Cryo-EM reconstructions of all four splice variants, together with SAXS and negative stain EM, reveal compacted dimers for each, with variant-specific dimeric arrangements. This results in specific trans-cellular interactions, as tested in cell clustering and stripe assays. The compact conformations provide a structural basis for teneurin homo- and heterophilic interactions. Altogether, our findings demonstrate how alternative splicing results in rearrangements of the dimeric subunits, influencing neuronal recognition and likely circuit wiring.

Sexual dimorphism in the social behaviour of Cntnap2-null mice correlates with disrupted synaptic connectivity and increased microglial activity in the anterior cingulate cortex.

A biological understanding of the apparent sex bias in autism is lacking. Here we have identified Cntnap2 KO mice as a model system to help better understand this dimorphism. Using this model, we observed social deficits in juvenile male KO mice only. These male-specific social deficits correlated with reduced spine densities of Layer 2/3 and Layer 5 pyramidal neurons in the Anterior Cingulate Cortex, a forebrain region prominently associated with the control of social behaviour. Furthermore, in male KO mice, microglia showed an increased activated morphology and phagocytosis of synaptic structures compared to WT mice, whereas no differences were seen in female KO and WT mice. Our data suggest that sexually dimorphic microglial activity may be involved in the aetiology of ASD, disrupting the development of neural circuits that control social behaviour by overpruning synapses at a developmentally critical period.

Cardiorespiratory kinetics in exercise physiology: estimates and predictions using randomized changes in work rate.

PURPOSE: Kinetics of cardiorespiratory parameters (CRP) in response to work rate (WR) changes are evaluated by pseudo-random binary sequences (PRBS testing). In this study, two algorithms were applied to convert responses from PRBS testing into appropriate impulse responses to predict steady states values and responses to incremental increases in exercise intensity. METHODS: 13 individuals (age: 41 ± 9 years, BMI: 23.8 ± 3.7 kg m-2), completing an exercise test protocol, comprising a section of randomized changes of 30 W and 80 W (PRBS), two phases of constant WR at 30 W and 80 W and incremental WR until subjective fatigue, were included in the analysis. Ventilation ([Formula: see text]), O2 uptake ([Formula: see text]), CO2 output ([Formula: see text]) and heart rate (HR) were monitored. Impulse responses were calculated in the time domain and in the frequency domain from the cross-correlations of WR and the respective CRP. RESULTS: The algorithm in the time domain allows better prediction for [Formula: see text] and [Formula: see text], whereas for [Formula: see text] and HR the results were similar for both algorithms. Best predictions were found for [Formula: see text] and HR with higher (3-4%) 30 W steady states and lower (1-4%) values for 80 W. Tendencies were found in the residuals between predicted and measured data. CONCLUSION: The CRP kinetics, resulting from PRBS testing, are qualified to assess steady states within the applied WR range. Below the ventilatory threshold, [Formula: see text] and HR responses to incrementally increasing exercise intensities can be sufficiently predicted.

Confinement, partial sleep deprivation and defined physical activity-influence on cardiorespiratory regulation and capacity.

INTRODUCTION: Adequate cardiorespiratory fitness is of utmost importance during spaceflight and should be assessable via moderate work rate intensities, e.g., using kinetics parameters. The combination of restricted sleep, and defined physical exercise during a 45-day simulated space mission is expected to slow heart rate (HR) kinetics without changes in oxygen uptake ([Formula: see text]) kinetics. METHODS: Overall, 14 crew members (9 males, 5 females, 37 ± 7 yrs, 23.4 ± 3.5 kg m-2) simulated a 45-d-mission to an asteroid. During the mission, the sleep schedule included 5 nights of 5 h and 2 nights of 8 h sleep. The crew members were tested on a cycle ergometer, using pseudo-random binary sequences, changing between 30 and 80 W on day 8 before (MD-8), day 22 (MD22) and 42 (MD42) after the beginning and day 4 (MD + 4) following the end of the mission. Kinetics information was assessed using the maxima of cross-correlation functions (CCFmax). Higher CCFmax indicates faster responses. RESULTS: CCFmax(HR) was significantly (p = 0.008) slower at MD-8 (0.30 ± 0.06) compared with MD22 (0.36 ± 0.06), MD42 (0.38 ± 0.06) and MD + 4 (0.35 ± 0.06). Mean HR values during the different work rate steps were higher at MD-8 and MD + 4 compared to MD22 and MD42 (p < 0.001). DISCUSSION: The physical training during the mission accelerated HR kinetics, but had no impact on mean HR values post mission. Thus, HR kinetics seem to be sensitive to changes in cardiorespiratory fitness and may be a valuable parameter to monitor fitness. Kinetics and capacities adapt independently in response to confinement in combination with defined physical activity and sleep.

The GTPase Arl8B Plays a Principle Role in the Positioning of Interstitial Axon Branches by Spatially Controlling Autophagosome and Lysosome Location.

Interstitial axon branching is an essential step during the establishment of neuronal connectivity. However, the exact mechanisms on how the number and position of branches are determined are still not fully understood. Here, we investigated the role of Arl8B, an adaptor molecule between lysosomes and kinesins. In chick retinal ganglion cells (RGCs), downregulation of Arl8B reduces axon branch density and shifts their location more proximally, while Arl8B overexpression leads to increased density and more distal positions of branches. These alterations correlate with changes in the location and density of lysosomes and autophagosomes along the axon shaft. Diminishing autophagy directly by knock-down of atg7, a key autophagy gene, reduces branch density, while induction of autophagy by rapamycin increases axon branching, indicating that autophagy plays a prominent role in axon branch formation. In vivo, local inactivation of autophagy in the retina using a mouse conditional knock-out approach disturbs retino-collicular map formation which is dependent on the formation of interstitial axon branches. These data suggest that Arl8B plays a principal role in the positioning of axon branches by spatially controlling autophagy, thus directly controlling formation of neural connectivity in the brain.SIGNIFICANCE STATEMENT The formation of interstitial axonal branches plays a prominent role in numerous places of the developing brain during neural circuit establishment. We show here that the GTPase Arl8B controls density and location of interstitial axon branches, and at the same time controls also density and location of the autophagy machinery. Upregulation or downregulation of autophagy in vitro promotes or inhibits axon branching. Local disruption of autophagy in vivo disturbs retino-collicular mapping. Our data suggest that Arl8B controls axon branching by controlling locally autophagy. This work is one of the first reports showing a role of autophagy during early neural circuit development and suggests that autophagy in general plays a much more prominent role during brain development than previously anticipated.

Adaptation of Systemic and Pulmonary Circulation to Acute Changes in Gravity and Body Position.

INTRODUCTION: Changes in gravity or body position provoke changes in hydrostatic pressure in the arterial system and in venous return. Potential asymmetries between left (QLV) and right ventricular (QRV) cardiac output during transient gravity changes were investigated. It was hypothesized that blood volume is temporarily stored in the pulmonary vessels, with amount and duration depending on the level and directions of gravity.METHODS: Eight healthy, male subjects (32 ± 3 yr, 182 ± 7 cm, 82 ± 6 kg) were tested on a tilt seat (TS), in a long arm human centrifuge (laHC), and during parabolic flights (PF). The gravitational changes during PF were reconstructed by changing gravity in a laHC and different body positions on a TS. All participants were tested in the seated, resting position. Heart rate and blood pressure were recorded continuously and QLV was calculated, applying the Modelflow Algorithm. Gas exchange was measured breath-by-breath. QRV was calculated from these data according to the Fick Principle. Four sequences were superimposed and analyzed by ANOVA with the factors Time, Ventricle (QRV, QLV), and Mode (TS, PF, laHC).RESULTS: After reductions in gravity QRV and QLV were transiently desynchronized. ANOVA showed no main effect for Mode, but significant changes were found for Time and Ventricle and all interactions.DISCUSSION: Phases of reduced gravity seem to lead to transiently increased storage of blood volume inside the pulmonary vascular system. A more detailed understanding of these mechanisms might help to describe the compliance of the pulmonary vascular system in diseases of the pulmonary circulation.Hoffmann U, Koschate J, Appell Coriolano H-J, Drescher U, Thieschäfer L, Dumitrescu D, Werner A. Adaptation of systemic and pulmonary circulation to acute changes in gravity and body position. Aerosp Med Hum Perform. 2019; 90(8):688-695.

Cardiorespiratory kinetics: comparisons between athletes with different training habits.

PURPOSE: Fast muscular oxygen uptake ([Formula: see text]) kinetics are limiting factors for high exercise capacities. It is hypothesized that [Formula: see text] and heart rate (HR) kinetics would be faster in individuals, performing long-distance endurance training (CONT) compared with athletes performing predominantly interval-based sports (INT). METHODS: 17 subjects (INT: n = 7, 24 ± 5 years, 183 ± 7 cm, 85 ± 10 kg, 6 ± 3 h of training per week, CONT: n = 10, 37 ± 7 years, 175 ± 9 cm, 69 ± 10 kg, 6 ± 3 h of training per week) completed a treadmill work rate (WR) protocol with pseudo-randomized WR changes with velocities of 6.5 and 9.5 km h-1. [Formula: see text]O2musc and the respective kinetics were estimated from the measured pulmonary oxygen uptake and HR combined with a circulatory model. Kinetics information were calculated using time series analysis. Higher maxima of the cross-correlation function (CCF) of WR and the respective parameter ([Formula: see text], HR) indicate faster kinetics responses. RESULTS: The kinetics of HR (INT: 0.23 ± 0.04 vs. CONT: 0.42 ± 0.18; P = 0.001), [Formula: see text]O2pulm (0.30 ± 0.05 vs. 0.53 ± 0.20; P = 0.005) and [Formula: see text]O2musc (0.31 ± 0.06 vs. 0.53 ± 0.16; P = 0.005) were significantly slower in INT compared with the CONT athletes. CONCLUSIONS: It seems that at least in the long-term CONT exercise, training without the need of changing intensities is favorable for fast [Formula: see text]O2 and HR kinetics compared with INT exercise including frequently changing intensities.

Impact of venous return on pulmonary oxygen uptake kinetics during dynamic exercise: in silico time series analyses from muscles to lungs.

The aim of the present study was to investigate whether a single-compartment (SCM) and a multi-compartment (MCM) venous return model will produce significantly different time-delaying and distortive effects on pulmonary oxygen uptake (V̇o2pulm) responses with equal cardiac outputs (Q̇) and muscle oxygen uptake (V̇o2musc) inputs. For each model, 64 data sets were simulated with alternating Q̇ and V̇o2musc kinetics-time constants (τ) ranging from 10 to 80 s-as responses to pseudorandom binary sequence work rate (WR) changes. Kinetic analyses were performed by using cross-correlation functions (CCFs) between WR with V̇o2pulm and V̇o2musc. Higher maxima of the CCF courses indicate faster system responses-equal to smaller τ values of the variables of interest (e.g., τV̇o2musc). The models demonstrated a highly significant relationship for the resulting V̇o2pulm responses ( r = 0.976, P < 0.001, n = 64). Both models showed significant differences between V̇o2pulm and V̇o2musc kinetics for τV̇o2musc ranging from 10 to 30 s ( P < 0.05 each). In addition, a significant difference in V̇o2pulm kinetics ( P < 0.05) between the models was observed for very fast V̇o2musc kinetics (τ = 10 s). The combinations of fast Q̇ dynamics and slow V̇o2musc kinetics yield distinct deviations in the resultant V̇o2pulm responses compared with V̇o2musc kinetics. Therefore, the venous return models should be used with care and caution if the aim is to infer V̇o2musc by means of V̇o2pulm kinetics. Finally, the resultant V̇o2pulm responses seem to be complex and most likely unpredictable if no cardiodynamic measurements are available in vivo. NEW & NOTEWORTHY A single-compartment and a multi-compartment venous return model were tested to see whether they result in different pulmonary oxygen uptake (V̇o2pulm) kinetics from equal cardiac output and muscle oxygen uptake (V̇o2musc) kinetics. To infer V̇o2musc kinetics by means of V̇o2pulm kinetics, both models should only be used for V̇o2musc time constants ranging from 40 to 80 s. The resultant V̇o2pulm responses seem to be complex and most likely unpredictable if no cardiodynamic measurements are available.

Multitasking During Simulated Car Driving: A Comparison of Young and Older Persons.

Human multitasking is typically studied by repeatedly presenting two tasks, either sequentially (task switch paradigms) or overlapping in time (dual-task paradigms). This is different from everyday life, which typically presents an ever-changing sequence of many different tasks. Realistic multitasking therefore requires an ongoing orchestration of task switching and dual-tasking. Here we investigate whether the age-related decay of multitasking, which has been documented with pure task-switch and pure dual-task paradigms, can also be quantified with a more realistic car driving paradigm. 63 young (20-30 years of age) and 61 older (65-75 years of age) participants were tested in an immersive driving simulator. They followed a car that occasionally slowed down and concurrently executed a mixed sequence of loading tasks that differed with respect to their sensory input modality, cognitive requirements and motor output channel. In two control conditions, the car-following or the loading task were administered alone. Older participants drove more slowly, more laterally and more variably than young ones, and this age difference was accentuated in the multitask-condition, particularly if the loading task took participants' gaze and attention away from the road. In the latter case, 78% of older drivers veered off the road and 15% drove across the median. The corresponding values for young drivers were 40% and 0%, respectively. Our findings indicate that multitasking deteriorates in older age not only in typical laboratory paradigms, but also in paradigms that require orchestration of dual-tasking and task switching. They also indicate that older drivers are at a higher risk of causing an accident when they engage in a task that takes gaze and attention away from the road.

Cognitive-Motor Interference in an Ecologically Valid Street Crossing Scenario.

Laboratory-based research revealed that gait involves higher cognitive processes, leading to performance impairments when executed with a concurrent loading task. Deficits are especially pronounced in older adults. Theoretical approaches like the multiple resource model highlight the role of task similarity and associated attention distribution problems. It has been shown that in cases where these distribution problems are perceived relevant to participant's risk of falls, older adults prioritize gait and posture over the concurrent loading task. Here we investigate whether findings on task similarity and task prioritization can be transferred to an ecologically valid scenario. Sixty-three younger adults (20-30 years of age) and 61 older adults (65-75 years of age) participated in a virtual street crossing simulation. The participants' task was to identify suitable gaps that would allow them to cross a simulated two way street safely. Therefore, participants walked on a manual treadmill that transferred their forward motion to forward displacements in a virtual city. The task was presented as a single task (crossing only) and as a multitask. In the multitask condition participants were asked, among others, to type in three digit numbers that were presented either visually or auditorily. We found that for both age groups, street crossing as well as typing performance suffered under multitasking conditions. Impairments were especially pronounced for older adults (e.g., longer crossing initiation phase, more missed opportunities). However, younger and older adults did not differ in the speed and success rate of crossing. Further, deficits were stronger in the visual compared to the auditory task modality for most parameters. Our findings conform to earlier studies that found an age-related decline in multitasking performance in less realistic scenarios. However, task similarity effects were inconsistent and question the validity of the multiple resource model within ecologically valid scenarios.

Effect of acute ambient temperature exposure on cardio-pulmonary and respiratory kinetics in men.

AIM: The goal of the study was to compare the kinetic responses of heart rate (HR) and pulmonary (V̇O2pulm) and muscular (V̇O2musc) oxygen uptake during dynamic leg exercise across different acute ambient temperature conditions in a climatic chamber. METHODS: Thirteen physically healthy, active, male volunteers demonstrated pseudorandom binary sequence (PRBS) work rate (WR) changes between 30 and 80 W at 15 °C, 25 °C and 35 °C, respectively. HR was measured beat-to-beat using an echocardiogram and V̇O2pulm by breath-by-breath gas exchange; V̇O2musc estimations were assessed by applying a circulatory model and cross-correlation functions. RESULTS: No significant differences were observed across the various temperature conditions in each case for HR, V̇O2pulm or V̇O2musc kinetics (p > 0.05). Baroreflex regulation based on HR kinetics does not seem to be influenced between ambient temperatures of 15 °C and 35 °C during dynamic exercise. CONCLUSIONS: The results imply that ambient temperatures of 15 °C, 25 °C and 35 °C have no effect on HR, V̇O2pulm or V̇O2musc kinetics during dynamic moderate exercise. The applied approach may be of interest for assessments of the cardio-pulmonary and respiratory health statuses of individuals working or performing sports in extreme temperature environments. Furthermore, differentiation between systemic (e.g. cardio-dynamic: HR) and specific (e.g. exercising tissues: V̇O2musc) determinants of the relevant physiological systems may improve the evaluation of an individual's health status.

Non-invasive estimation of muscle oxygen uptake kinetics with pseudorandom binary sequence and step exercise responses.

PURPOSE: The aim of the study was to test for significant differences in non-invasively estimated muscle oxygen uptake ([Formula: see text]) kinetics, assessed by a square-wave exercise protocol (STEP) as well as by a time series approach with pseudorandom binary sequence (PRBS) work rate (WR) changes. METHODS: Seventeen healthy and active individuals (10 women, 7 men; 23 ± 2 years old; height 175 ± 11 cm; body mass 73 ± 14 kg [mean ± SD]) completed five repetitions of WR transitions from 30 to 80 W for the STEP approach and two sequences of pseudorandom binary WR changes between 30 and 80 W for the PRBS approach. Pulmonary oxygen uptake ([Formula: see text]) was measured breath by breath. [Formula: see text] kinetics were estimated during phase II [Formula: see text] in the STEP approach and during the pseudorandom binary sequence WR changes in the PRBS approach. RESULTS: No significant differences were observed between different models of the STEP and the PRBS approach for estimation of [Formula: see text] kinetics (p > 0.05). In addition, a very high variability between the models was determined for [Formula: see text] kinetics [mean time constants (τ) difference: - 2.5 ± 11.4 s]. A significant correlation for τ of [Formula: see text] between the STEP approach with experimentally determined phase I [Formula: see text] lengths and the PRBS approach was noticed (r = 0.536; p < 0.05). CONCLUSIONS: Both approaches (STEP and PRBS) are not significantly different for estimating the [Formula: see text] kinetics, but the very high variability impairs the predictability between the models. However, the determination of the length of phase I [Formula: see text] should be as appropriate as possible because predefined duration lengths can result in overestimations in [Formula: see text] kinetics.

Heart Rate and Oxygen Uptake Kinetics in Type 2 Diabetes Patients - A Pilot Study on the Influence of Cardiovascular Medication on Regulatory Processes.

The aim of this pilot study was to investigate whether there are differences in heart rate and oxygen uptake kinetics in type 2 diabetes patients, considering their cardiovascular medication. It was hypothesized that cardiovascular medication would affect heart rate and oxygen uptake kinetics and that this could be detected using a standardized exercise test. 18 subjects were tested for maximal oxygen uptake. Kinetics were measured in a single test session with standardized, randomized moderate-intensity work rate changes. Time series analysis was used to estimate kinetics. Greater maxima in cross-correlation functions indicate faster kinetics. 6 patients did not take any cardiovascular medication, 6 subjects took peripherally acting medication and 6 patients were treated with centrally acting medication. Maximum oxygen uptake was not significantly different between groups. Significant main effects were identified regarding differences in muscular oxygen uptake kinetics and heart rate kinetics. Muscular oxygen uptake kinetics were significantly faster than heart rate kinetics in the group with no cardiovascular medication (maximum in cross-correlation function of muscular oxygen uptake vs. heart rate; 0.32±0.08 vs. 0.25±0.06; p=0.001) and in the group taking peripherally acting medication (0.34±0.05 vs. 0.28±0.05; p=0.009) but not in the patients taking centrally acting medication (0.28±0.05 vs. 0.30±0.07; n.s.). It can be concluded that regulatory processes for the achievement of a similar maximal oxygen uptake are different between the groups. The used standardized test provided plausible results for heart rate and oxygen uptake kinetics in a single measurement session in this patient group.

On the role of ephrinA2 in auditory function.

Recent findings suggest that the manipulation of the EphA/ephrinA system can improve hearing threshold sensitivity in the auditory system (Yates et al., 2014). These results appear to open-up the possibility that pharmacological manipulation of this system could lead to the development of treatments to cure some types of hearing loss. As a first step towards this goal, we have performed a further series of auditory brainstem evoked potential recordings on ephrinA2 homozygous knockout mice and their wildtype littermates in order to replicate the previously reported findings. However, we found that ephrinA2 knockout mice had auditory threshold sensitivity for click and 3-42 kHz tone pip frequencies comparable to that of their wildtype littermates. Evoked potential wave amplitudes, latencies and inter-peak intervals were also comparable between ephrinA2 knockout mice and wild type control littermates. Thus in our experiments we could not replicate the findings of Yates et al. (2014). Whilst the EphA/ephrinA system may therefore play a role in the development of innervation of the cochlea and neural circuitry of the auditory brainstem, there appears to be a functional redundancy between members of this family such that loss of ephrinA2 function alone is insufficient to alter auditory function in the mouse.

Faster heart rate and muscular oxygen uptake kinetics in type 2 diabetes patients following endurance training.

Cardiorespiratory kinetics were analyzed in type 2 diabetes patients before and after a 12-week endurance exercise-training intervention. It was hypothesized that muscular oxygen uptake and heart rate (HR) kinetics would be faster after the training intervention and that this would be detectable using a standardized work rate protocol with pseudo-random binary sequences. The cardiorespiratory kinetics of 13 male sedentary, middle-aged, overweight type 2 diabetes patients (age, 60 ± 8 years; body mass index, 33 ± 4 kg·m-2) were tested before and after the 12-week exercise intervention. Subjects performed endurance training 3 times a week on nonconsecutive days. Pseudo-random binary sequences exercise protocols in combination with time series analysis were used to estimate kinetics. Greater maxima in cross-correlation functions (CCFmax) represent faster kinetics of the respective parameter. CCFmax of muscular oxygen uptake (pre-training: 0.31 ± 0.03; post-training: 0.37 ± 0.1, P = 0.024) and CCFmax of HR (pre-training: 0.25 ± 0.04; post-training: 0.29 ± 0.06, P = 0.007) as well as peak oxygen uptake (pre-training: 24.4 ± 4.7 mL·kg-1·min-1; post-training: 29.3 ± 6.5 mL·kg-1·min-1, P = 0.004) increased significantly over the course of the exercise intervention. In conclusion, kinetic responses to changing work rates in the moderate-intensity range are similar to metabolic demands occurring in everyday habitual activities. Moderate endurance training accelerated the kinetic responses of HR and muscular oxygen uptake. Furthermore, the applicability of the used method to detect these accelerations was demonstrated.

Target-independent ephrina/EphA-mediated axon-axon repulsion as a novel element in retinocollicular mapping.

EphrinAs and EphAs play critical roles during topographic map formation in the retinocollicular projection; however, their complex expression patterns in both the retina and superior colliculus (SC) have made it difficult to uncover their precise mechanisms of action. We demonstrate here that growth cones of temporal axons collapse when contacting nasal axons in vitro, and removing ephrinAs from axonal membranes by PI-PLC treatment abolishes this response. In conditional knockout mice, temporal axons display no major targeting defects when ephrinA5 is removed only from the SC, but substantial mapping defects were observed when ephrinA5 expression was removed from both the SC and from the retina, with temporal axons invading the target areas of nasal axons. Together, these data indicate that ephrinA5 drives repellent interactions between temporal and nasal axons within the SC, and demonstrates for the first time that target-independent mechanisms play an essential role in retinocollicular map formation in vivo.

BDNF promotes axon branching of retinal ganglion cells via miRNA-132 and p250GAP.

A crucial step in the development of the vertebrate visual system is the branching of retinal ganglion cell (RGC) axons within their target, the superior colliculus/tectum. A major player in this process is the neurotrophin brain-derived neurotrophic factor (BDNF). However, the molecular basis for the signaling pathways mediating BDNF action is less well understood. As BDNF exerts some of its functions by controlling the expression of microRNAs (miRNAs), we investigated whether miRNAs are also involved in BDNF-mediated retinal axon branching. Here, we demonstrate that the expression pattern of miRNA-132 in the retina is consistent with its involvement in this process, and that BDNF induces the upregulation of miRNA-132 in retinal cultures. Furthermore, in vitro gain-of-function and loss-of-function approaches in retinal cultures reveal that miRNA-132 mediates axon branching downstream of BDNF. A known target of miRNA-132 is the Rho family GTPase-activating protein, p250GAP. We find that p250GAP is expressed in RGC axons and mediates the effects of miRNA-132 in BDNF-induced branching. BDNF treatment or overexpression of miRNA-132 leads to a reduction in p250GAP protein levels in retinal cultures, whereas the overexpression of p250GAP abolishes BDNF-induced branching. Finally, we used a loss-of-function approach to show that miRNA-132 affects the maturation of RGC termination zones in the mouse superior colliculus in vivo, while their topographic targeting remains intact. Together, our data indicate that BDNF promotes RGC axon branching during retinocollicular/tectal map formation via upregulation of miRNA-132, which in turn downregulates p250GAP.

Protein tyrosine phosphatase receptor type O inhibits trigeminal axon growth and branching by repressing TrkB and Ret signaling.

Axonal branches of the trigeminal ganglion (TG) display characteristic growth and arborization patterns during development. Subsets of TG neurons express different receptors for growth factors, but these are unlikely to explain the unique patterns of axonal arborizations. Intrinsic modulators may restrict or enhance cellular responses to specific ligands and thereby contribute to the development of axon growth patterns. Protein tyrosine phosphatase receptor type O (PTPRO), which is required for Eph receptor-dependent retinotectal development in chick and for development of subsets of trunk sensory neurons in mouse, may be such an intrinsic modulator of TG neuron development. PTPRO is expressed mainly in TrkB-expressing (TrkB(+)) and Ret(+) mechanoreceptors within the TG during embryogenesis. In PTPRO mutant mice, subsets of TG neurons grow longer and more elaborate axonal branches. Cultured PTPRO(-/-) TG neurons display enhanced axonal outgrowth and branching in response to BDNF and GDNF compared with control neurons, indicating that PTPRO negatively controls the activity of BDNF/TrkB and GDNF/Ret signaling. Mouse PTPRO fails to regulate Eph signaling in retinocollicular development and in hindlimb motor axon guidance, suggesting that chick and mouse PTPRO have different substrate specificities. PTPRO has evolved to fine tune growth factor signaling in a cell-type-specific manner and to thereby increase the diversity of signaling output of a limited number of receptor tyrosine kinases to control the branch morphology of developing sensory neurons. The regulation of Eph receptor-mediated developmental processes by protein tyrosine phosphatases has diverged between chick and mouse.

Axonal ephrinA/EphA interactions, and the emergence of order in topographic projections.

In the classical view of axon guidance, neurons send out axons which are endowed with guidance receptors enabling them to find their (distant) target areas by an interaction with their ligands expressed in specific spatio-temporal patterns along their pathways and in their target area. However, this view has recently been confounded by more detailed analyses of, for example, the expression patterns of EphAs and ephrinAs in the retinotectal projection. Here ephrinA 'ligands' are expressed not only in the target area but also on the projecting RGC axons, and EphA 'receptors' not only on retinal ganglion cell (RGC) axons but also in the target area itself. This review describes the on-going functional characterisation of the surprising co-expression of ephrinAs and EphAs on retinal ganglion cell (RGC) axons and other cell types. It also investigates the function of ephrinAs as receptors and describes their interaction with co-receptors involved in mediating this function.

Localization of Cadm2a and Cadm3 proteins during development of the zebrafish nervous system.

Members of the Cadm/SynCAM/Necl/IGSF/TSLC family of cell adhesion molecules are known to have diverse functions during development of the nervous system, but information regarding their role during central nervous system (CNS) development in vivo is scarce. The rapid development of a relatively simple nervous system in larval zebrafish makes them a highly tractable model organism for studying gene function during nervous system development. An essential prerequisite for functional studies is a description of protein localization. To address this we have generated subtype-specific antibodies to two members of the zebrafish cell adhesion molecule family: cadm2a and cadm3. Using these novel antibodies we show that cadm3 and cadm2a are expressed throughout the nervous system of larval stage zebrafish. Particularly striking, and largely nonoverlapping expression of cadm2a and cadm3 is observed in the developing retina and spinal cord. Using in vitro binding assays we show that cadm2a and cadm3 bind heterophilically and preferentially to cadm1 and cadm4, respectively. These binding preferences are very similar to those seen for tetrapod Cadms but our study of protein localization suggests novel and diverse functions of cadms during nervous system development.