Lectin binding and gel secretion within Lorenzinian electroreceptors of Polyodon.

We imaged the carbohydrate-selective spatial binding of 8 lectins in the ampullary organs (AOs) of electroreceptors on the rostrum of freshwater paddlefish (Polyodon spathula), by fluorescence imaging and morphometry of frozen sections. A focus was candidate sites of secretion of the glycoprotein gel filling the lumen of AOs. The rostrum of Polyodon is an electrosensory appendage anterior of the head, covered with >50,000 AOs, each homologous with the ampulla of Lorenzini electroreceptors of marine rays and sharks. A large electrosensory neuroepithelium (EN) lines the basal pole of each AO's lumen in Polyodon; support cells occupy most (97%) of an EN's apical area, along with electrosensitive receptor cells. (1) Lectins WGA or SBA labeled the AO gel. High concentrations of the N-acetyl-aminocarbohydrate ligands of these lectins were reported in canal gel of ampullae of Lorenzini, supporting homology of Polyodon AOs. In cross sections of EN, WGA or SBA labeled cytoplasmic vesicles and organelles in support cells, especially apically, apparently secretory. Abundant phalloidin+ microvilli on the apical faces of support cells yielded the brightest label by lectins WGA or SBA. In parallel views of the apical EN surface, WGA labeled only support cells. We concluded that EN support cells massively secrete gel from their apical microvilli (and surface?), containing amino carbohydrate ligands of WGA or SBA, into the AO lumen. (2) Lectins RCA120 or ConA also labeled EN support cells, each differently. RCA120-fluorescein brightly labeled extensive Golgi tubules in the apical halves of EN cells. ConA did not label microvilli, but brightly labeled small vesicles throughout support cells, apparently non-secretory. (3) We demonstrated "sockets" surrounding the basolateral exteriors of EN receptor cells, as candidate glycocalyces. (4) We explored whether additional secretions may arise from non-EN epithelial cells of the interior ampulla wall. (5) Model: Gel is secreted mainly by support cells in the large EN covering each AO's basal pole. Secreted gel is pushed toward the pore, and out. We modeled gel velocity as increasing ~11x, going distally in AOs (toward the narrowed neck and pore), due to geometrical taper of the ampulla wall. Gel renewal and accelerated expulsion may defend against invasion of the AO lumen by microbes or small parasites. (6) We surveyed lectin labeling of accessory structures, including papilla cells in AO necks, striated ectoderm epidermis, and sheaths on afferent axons or on terminal glia.

Papilla cells may guard the entrance to ampullary organs of Polyodon electroreceptors.

We imaged 'papilla cells' in the skin pores of ampullary organs in electroreceptors on the rostrum of freshwater paddlefish (Polyodon spathula), by lectin labeling and fluorescence stack imaging of frozen sections. >50,000 ampullary organs, each 0.3-0.5 mm deep, are embedded in special skin covering the rostrum; their skin pores offer invasion routes into the body interior. We imaged numerous small papillas, of lengths 6-31 microns, lining the luminal surfaces of the narrowed neck and skin pore, at the entrance to each ampullary organ. Lectins WGA or SBA labeled their surfaces. Each papilla led to an oblong base containing a nucleus, embedded in the cuboidal epithelium of the neck wall; the papillas were anucleate. Our immunolabeling of junctions between papilla cells for ZO1, TJP2, occludin, tricellulin, phalloidin, or collagen-1 was consistent with zona occludens. We confirmed the two-part morphology of papilla cells by acute dissociation of individual neck epithelial cells (using two methods), which showed a conical papilla, angled (on some) relative to a nucleated base. We propose that papilla cells may be defensive in function, strategically located at the skin pore and neck to deter entry of pathogens into ampullary organs, including microbes or small parasites.

Large-Scale Convergence of Receptor Cell Arrays Onto Afferent Terminal Arbors in the Lorenzinian Electroreceptors of Polyodon.

Certain sensory receptors contain many transducers, converging onto few afferents. Convergence creates star-topology neural networks, of iterative parallel organization, that may yield special functional properties. We quantitated large-scale convergence in electroreceptors on the rostrum of preadult paddlefish, Polyodon spathula (Acipenseriforme vertebrates), and analyzed the afferent terminal branching underlying the convergence. From neurophysiological mapping, a recorded afferent innervated 23.3 ± 9.1 (range 6-45) ampullary organs, and innervated every ampullary organ within the receptive field's sharp boundary. Ampullary organs each contained ∼665 Lorenzinian receptor cells, from imaging and modeling. We imaged three serial types of afferent branching at electroreceptors, after immunofluorescent labeling for neurite filaments, glial sheaths, or nodal ion channels, or by DiI tracing. (i) Myelinated tree: Each of 3.08 ± 0.51 (2-4) parallel afferents from a cranial nerve (ALLn) entered a receptive field from deeper tissue, then branched into a laminar tree of large myelinated dendrites, parallel to the skin, that branched radially until ∼9 extremities with heminodes, which were candidate sites of spike encoders. (ii) Inline transition: Each myelinated extremity led distally into local unmyelinated arbors originating at inline branching structures covered by terminal (satellite) glia. The unmyelinated transition zones included globular afferent modules, 4-6 microns wide, from which erupted fine fascicles of parallel submicron neurites, a possibly novel type of neuronal branching. The neurite fascicles formed loose bundles projecting ∼105 microns distally to innervate local groups of ∼3 adjacent ampullary organs. (iii) Radial arbors: Receptor cells in an electrosensory neuroepithelium covering the basal pole of each ampullary organ were innervated by bouton endings of radial neurites, unmyelinated and submicron, forming a thin curviplanar lamina distal to the lectin+ basal lamina. The profuse radial neurites diverged from thicker (∼2 micron) basolateral trunks. Overall, an average Polyodon electroreceptor formed a star topology array of ∼9 sensor groups. Total convergence ratios were 15,495 ± 6,052 parallel receptor cells per afferent per mean receptive field, assuming 100% innervation. Large-scale convergence likely increases the signal-to-noise ratio (SNR) of stimulus encoding into spiking afferent output, increasing receiver sensitivity. Unmyelinated arbors may also regenerate and repair the afferent innervation of ampullary organs. LSID: urn:lsid:zoobank.org:act:09BCF04C-3C3C-4B6C-9DC9-A2BF43087369.

Quantitative measurement of lower limb mechanical alignment and coronal knee laxity in early flexion.

BACKGROUND: Non-invasive quantification of lower limb alignment using navigation technology is now possible throughout knee flexion owing to software developments. We report the precision and accuracy of a non-invasive system measuring mechanical alignment of the lower limb including coronal stress testing of the knee. METHODS: Twelve cadaveric limbs were tested with a commercial invasive navigation system against the non-invasive system. Coronal mechanical femorotibial (MFT) alignment was measured with no stress, then 15 Nm varus and valgus applied moments. Measurements were recorded at 10° intervals from extension to 90° flexion. At each flexion interval, coefficient of repeatability (CR) tested precision within each system, and limits of agreement (LOA) tested agreement between the two systems. Limits for CR & LOA were set at 3° based on requirements for surgical planning and evaluation. RESULTS: Precision was acceptable throughout flexion in all conditions of stress using the invasive system (CR ≤ 1.9°). Precision was acceptable using the non-invasive system from extension to 50° flexion (CR ≤ 2.4°), beyond which precision was unacceptable (> 3.4°). With no coronal stress applied, agreement remained acceptable from extension to 40° (LOA ≤ 2.4°), and when 15 Nm varus or valgus stress was applied agreement was acceptable from extension to 30° (LOA ≤ 2.9°). Higher angles of knee flexion had a negative impact on precision and accuracy. CONCLUSION & CLINICAL RELEVANCE: The non-invasive system provides reliable quantitative data in-vitro on coronal MFT alignment and laxity in the range relevant to assessment of collateral ligament injury, pre-operative planning of arthroplasty and flexion instability following arthroplasty. In-vivo validation should be performed.

Repeatability and accuracy of a non-invasive method of measuring internal and external rotation of the tibia.

PURPOSE: The ability to quantify rotational laxity of the knee would increase understanding of functional rotatory instability, identify the best treatment methods for soft tissue injury, and have a role in diagnosis of soft tissue injury. This study aimed to report the reliability, repeatability and precision of a non-invasive adaptation of image-free navigation technology by comparing with a validated invasive system used for computer-assisted surgery. METHODS: Twelve cadaveric lower limbs were tested with a commercial image-free navigation system using passive trackers secured by bone screws. They were then tested a non-invasive fabric-strap system. Manual application of torque was used consistent with clinical examination to rotate the tibia to the end of internal rotation and external rotation range. Measurements were taken at 10° intervals from full extension to 90° flexion, and protocol was repeated twice using each system. Intraclass correlation coefficient (ICC) was used to reflect reliability of measurements. At each flexion interval, coefficient of repeatability (CR) was calculated for each system, and limits of agreement (LOA) were used to reflect agreement between the systems. RESULTS: The results for internal and external rotation were combined throughout flexion: ICC invasive; 0.94 (0.86-0.99), non-invasive; 0.92 (0.7-0.99), CR invasive; 2.4° (1.3-4.8°), non-invasive; 3.5° (1.8-6.6), LOA; 8.2° (4.3-13.5). CONCLUSION: Non-invasive optical tracker fixation gives improved agreement with a validated method of measurement compared with devices measuring tibial rotation by foot position. This system gives the added possibility of dynamic, weight-bearing testing in the clinically important range of 0°-30° knee flexion without the need for any limb restraint.

Review of the evidence: surgical management of 4th and 5th tarsometatarsal joint osteoarthritis.

Osteoarthritis of the lateral tarsometatarsal joints is less common than that which is seen in the 1st-3rd tarsometatarsal joints. Despite a suspected increase in incidence of tarsometatarsal arthritis and consequently the burden of disability and economic impact, guidelines for treatment and decision making remain scarce. When conservative treatment fails, lateral column osteoarthritis can severely limit a patient's mobility, lifestyle, and present a difficult management problem for the foot and ankle specialist. Evidence for the surgical techniques used in treatment of lateral column osteoarthritis is limited and sporadic within the literature. This article aims to summarise and compare the evidence for these surgical management options. This article looks at aetiology and epidemiology, with a summary of the biomechanics of the region and a comprehensive review of the literature regarding surgical treatment options.

Characteristic effects of stochastic oscillatory forcing on neural firing: analytical theory and comparison to paddlefish electroreceptor data.

Stochastic signals with pronounced oscillatory components are frequently encountered in neural systems. Input currents to a neuron in the form of stochastic oscillations could be of exogenous origin, e.g. sensory input or synaptic input from a network rhythm. They shape spike firing statistics in a characteristic way, which we explore theoretically in this report. We consider a perfect integrate-and-fire neuron that is stimulated by a constant base current (to drive regular spontaneous firing), along with Gaussian narrow-band noise (a simple example of stochastic oscillations), and a broadband noise. We derive expressions for the nth-order interval distribution, its variance, and the serial correlation coefficients of the interspike intervals (ISIs) and confirm these analytical results by computer simulations. The theory is then applied to experimental data from electroreceptors of paddlefish, which have two distinct types of internal noisy oscillators, one forcing the other. The theory provides an analytical description of their afferent spiking statistics during spontaneous firing, and replicates a pronounced dependence of ISI serial correlation coefficients on the relative frequency of the driving oscillations, and furthermore allows extraction of certain parameters of the intrinsic oscillators embedded in these electroreceptors.

Identifying temporal codes in spontaneously active sensory neurons.

The manner in which information is encoded in neural signals is a major issue in Neuroscience. A common distinction is between rate codes, where information in neural responses is encoded as the number of spikes within a specified time frame (encoding window), and temporal codes, where the position of spikes within the encoding window carries some or all of the information about the stimulus. One test for the existence of a temporal code in neural responses is to add artificial time jitter to each spike in the response, and then assess whether or not information in the response has been degraded. If so, temporal encoding might be inferred, on the assumption that the jitter is small enough to alter the position, but not the number, of spikes within the encoding window. Here, the effects of artificial jitter on various spike train and information metrics were derived analytically, and this theory was validated using data from afferent neurons of the turtle vestibular and paddlefish electrosensory systems, and from model neurons. We demonstrate that the jitter procedure will degrade information content even when coding is known to be entirely by rate. For this and additional reasons, we conclude that the jitter procedure by itself is not sufficient to establish the presence of a temporal code.

Sensory coding in oscillatory electroreceptors of paddlefish.

Coherence and information theoretic analyses were applied to quantitate the response properties and the encoding of time-varying stimuli in paddlefish electroreceptors (ERs), studied in vivo. External electrical stimuli were Gaussian noise waveforms of varied frequency band and strength, including naturalistic waveforms derived from zooplankton prey. Our coherence analyses elucidated the role of internal oscillations and transduction processes in shaping the 0.5-20 Hz best frequency tuning of these electroreceptors, to match the electrical signals emitted by zooplankton prey. Stimulus-response coherence fell off above approximately 20 Hz, apparently due to intrinsic limits of transduction, but was detectable up to 40-50 Hz. Aligned with this upper fall off was a narrow band of intense internal noise at ∼25 Hz, due to prominent membrane potential oscillations in cells of sensory epithelia, which caused a narrow deadband of external insensitivity. Using coherence analysis, we showed that more than 76% of naturalistic stimuli of weak strength, ∼1 µV∕cm, was linearly encoded into an afferent spike train, which transmitted information at a rate of ∼30 bits∕s. Stimulus transfer to afferent spike timing became essentially nonlinear as the stimulus strength was increased to induce bursting firing. Strong stimuli, as from nearby zooplankton prey, acted to synchronize the bursting responses of afferents, including across populations of electroreceptors, providing a plausible mechanism for reliable information transfer to higher-order neurons through noisy synapses.

Coherent stochastic oscillations enhance signal detection in spiking neurons.

We study the effect of noisy oscillatory input on the signal discrimination by spontaneously firing neurons. Using analytically tractable model, we contrast signal detection in two situations: (i) when the neuron is driven by coherent oscillations and (ii) when the coherence of oscillations is destroyed. Analytical calculations revealed a region in the parameter space of the model where oscillations act to reduce the variability of neuronal firing and to enhance the discriminability of weak signals. These analytical results are employed to unveil a possible role of coherent oscillations in peripheral electrosensory system of paddlefish in improvement of detection of weak stimuli. The proposed mechanism may be relevant to a wide range of phenomena involving coherently driven oscillators.

Response clustering in transient stochastic synchronization and desynchronization of coupled neuronal bursters.

We studied the transient dynamics of synchronized coupled neuronal bursters subjected to repeatedly applied stimuli, using a hybrid neuroelectronic system of paddlefish electroreceptors. We show experimentally that the system characteristically undergoes poststimulus transients, in which the relative phases of the oscillators may be grouped in several clusters, traversing alternate phase trajectories. These signature transient dynamics can be detected and characterized quantitatively using specific statistical measures based on a stochastic approach to transient oscillator responses.

Noise-induced transition to bursting in responses of paddlefish electroreceptor afferents.

The response properties of ampullary electroreceptors of paddlefish, Polyodon spathula, were studied in vivo, as single-unit afferent responses to external electrical stimulation with varied intensities of several types of noise waveforms, all Gaussian and zero-mean. They included broadband white noise, Ornstein-Uhlenbeck noise, low- or high-frequency band-limited noise, or natural noise recorded from swarms of Daphnia zooplankton prey, or from individual prey. Normally the afferents fire spontaneously in a tonic manner, which is actually quasiperiodic due to embedded oscillators. 1) Weak noise stimuli increased the variability of afferent firing, but it remained tonic. 2) In contrast, stimulation with less-weak broadband noise led to a qualitative change of the firing patterns, to parabolic bursting, even though the mean firing rate was scarcely affected. 3) The transition to afferent bursting was marked by the development of two well-separated timescales: the fast frequency of spiking inside bursts at

Measuring direction in the coupling of biological oscillators: a case study for electroreceptors of paddlefish.

Recently developed methods for estimating directionality in the coupling between oscillators were tested on experimental time series data from electroreceptors of paddlefish, because each electroreceptor contains two distinct types of noisy oscillators. One type of oscillator is in the sensory epithelia, and another type is in the terminals of afferent neurons. Based on morphological organization and our previous work, we expected unidirectional coupling, whereby epithelial oscillations synaptically influence the spiking oscillators of afferent neurons. Using directionality analysis we confirmed unidirectional coupling of oscillators embedded in electroreceptors. We studied the performance of directionality algorithms for decreasing length of data. Also, we experimentally varied the strength of oscillator coupling, to test the effect of coupling strength on directionality algorithms.

Models of stochastic biperiodic oscillations and extended serial correlations in electroreceptors of paddlefish.

Two types of minimal models were used to study stochastic oscillations in sensory receptors composed of two coupled oscillators, as in the electroreceptors of paddlefish. They have populations of cells in sensory epithelia undergoing approximately 26 Hz oscillations. These are coupled unidirectionally via synaptic excitation to a few afferent neurons, each of whose terminal contains a 30-70 Hz oscillator, expressed as a dominant peak in the power spectra of spontaneous afferent firing, corresponding to the mean firing rate. The two distinct types of internal noisy oscillators result in stochastic biperiodic firing patterns of the primary afferent sensory neurons. However, the functions of the oscillations have remained elusive, motivating this study. The models we used here are based on the circle map, or on the Ermentraut-Koppell canonical phase (theta neuron) model. Parameters were chosen according to experimental data. We used the models to demonstrate that the presence of epithelial oscillations leads to extended negative correlations of afferent interspike intervals, and to show that the correlation structure depends crucially on the ratio of the afferent to epithelial oscillation frequencies, being most pronounced when this ratio is close to 2, as observed in experiments. Our studies of stochastic versions of these models are of general interest for a wide range of coupled excitable systems, especially for understanding the functional roles of noisy oscillations in auditory and other types of "hair cell-primary afferent" sensory receptors.

Information processing in noisy burster models of sensory neurons.

Processing of external stimuli by sensory neurons often involves bursting, when epochs of fast firing alternate with intervals of quiescence. In particular, sensory neurons of electroreceptors in paddlefish (Polyodon spathula) undergo bursting when stimulated externally with broad-band noise, but otherwise fire spontaneously in a quasiperiodic tonic manner. We use a simple phenomenological model for noise-induced bursting to quantify analytically, by means of the Kullback entropy and Fisher information, the gain in information transfer and electroreceptor sensitivity for external noisy stimuli. A good agreement between theoretical predictions, numerical simulations and experimental data is shown.

Two distinct types of noisy oscillators in electroreceptors of paddlefish.

Our computational analyses and experiments demonstrate that ampullary electroreceptors in paddlefish (Polyodon spathula) contain 2 distinct types of continuously active noisy oscillators. The spontaneous firing of afferents reflects both rhythms, and as a result is stochastically biperiodic (quasiperiodic). The first type of oscillator resides in the sensory epithelia, is recorded as approximately 26 Hz and +/-70 microV voltage fluctuations at the canal skin pores, and gives rise to a noisy peak at f(e) approximately 26 Hz in power spectra of spontaneous afferent firing. The second type of oscillator resides in afferent terminals, is seen as a noisy peak at f(a) approximately 30-70 Hz that dominates the power spectra of spontaneous afferent firing, and corresponds to the mean spontaneous firing rate. Sideband peaks at frequencies of f(a) +/- f(e) are consistent with epithelia-to-afferent unidirectional synaptic coupling or, alternatively, nonlinear mixing of the 2 oscillatory processes. External stimulation affects the frequency of only the afferent oscillator, not the epithelial oscillators. Application of temperature gradients localized the f(e) and f(a) oscillators to different depths below the skin. Having 2 distinct types of internal oscillators is a novel form of organization for peripheral sensory receptors, of relevance for other hair cell sensory receptors.

Synchronization of noise-induced bursts in noncoupled sensory neurons.

We report experimental observation of phase synchronization in an array of nonidentical noncoupled noisy neuronal oscillators, due to stimulation with external noise. The synchronization derives from a noise-induced qualitative change in the firing pattern of single neurons, which changes from a quasiperiodic to a bursting mode. We show that at a certain noise intensity the onsets of bursts in different neurons become synchronized, even though the number of spikes inside the bursts may vary for different neurons. We demonstrate this effect both experimentally for the electroreceptor afferents of paddlefish, and numerically for a canonical phase model, and characterize it in terms of stochastic synchronization.

Behavioral stochastic resonance: how the noise from a Daphnia swarm enhances individual prey capture by juvenile paddlefish.

Zooplankton emit weak electric fields into the surrounding water that originate from their own muscular activities associated with swimming and feeding. Juvenile paddlefish prey upon single zooplankton by detecting and tracking these weak electric signatures. The passive electric sense in this fish is provided by an elaborate array of electroreceptors, Ampullae of Lorenzini, spread over the surface of an elongated rostrum. We have previously shown that the fish use stochastic resonance to enhance prey capture near the detection threshold of their sensory system. However, stochastic resonance requires an external source of electrical noise in order to function. A swarm of plankton, for example Daphnia, can provide the required noise. We hypothesize that juvenile paddlefish can detect and attack single Daphnia as outliers in the vicinity of the swarm by using noise from the swarm itself. From the power spectral density of the noise plus the weak signal from a single Daphnia, we calculate the signal-to-noise ratio, Fisher information and discriminability at the surface of the paddlefish's rostrum. The results predict a specific attack pattern for the paddlefish that appears to be experimentally testable.