Intracranial portion of the trochlear nerve and dorsal oblique muscle composition in dog: a structural and ultrastructural study

In the present investigation the right intracranial portion of the trochlear nerves and dorsal oblique muscle of the right ocular globe were removed from six adult dogs and analyzed by light and electron microscopy. Unmyelinated fibers were observed in the analyzed nerves. The number, diameter, area, and density of myelinated fibers were determined, as were corresponding axon area and diameter and myelin sheath thickness. Frequency histograms of myelin sheath thickness and fiber size show a bimodal distribution with a similar proportion of large and small fibers. Muscle samples were taken from the central portion of the muscle belly, subsequently frozen, cut, and stained with m-ATPase at pH 4.6. Fibers were classified as Type 1 or Type 2 according to their reaction to the m-ATPase and detailed morphologic and morphometric studies were made. The muscles showed two clearly distinct layers, a central layer and a peripheral layer, chiefly composed of Type 2 fibers. The fibers in the central layer were larger in size than those in the peripheral layer.     

Dual video microscopic imaging of membrane potential and cytosolic calcium of immunoidentified embryonic rat cortical cells

Membrane potential (MP) and cytosolic Ca2+ (Ca2+(c)) constitute important components involved in the physiological regulation of a myriad of cell functions in eukaryotic organisms. In particular, during development of the central nervous system, both properties are thought to be important in the regulation of cell cycle, cell migration, cell differentiation, cell-cell communication, and naturally occurring cell death. However, obtaining insight into the precise relationship between these two parameters of cell function is relatively limited either by technical difficulties inherent in using electrical recordings of membrane properties in conjunction with optical imaging of single cells or by employing optical imaging of either one or another property alone. Here, we describe in detail a novel strategy to record changes in both MP and Ca2+(c) from many intact single cells in a noninvasive manner using digital video microscopy. This method involves double-loading the cells with voltage- and calcium-sensitive fluorescent indicator dyes, green oxonol, and fura-2, which can be sequentially excited with a mercury arc lamp filtered at appropriate wavelengths and their resulting emissions can be captured with an intensified charged-coupled device camera at 1-s intervals. As an example of the utility of dual-recording strategy, we present data on a distinct functional expression of excitable membrane and cytoplasmic calcium properties in proliferating and differentiating embryonic rat cerebral cortical cells.     

Tissue-specific neuro-glia interactions determine neurite differentiation in ganglion cells

Guided formation and extension of axons versus dendrites is considered crucial for structuring the nervous system. In the chick visual system, retinal ganglion cells (RGCs) extend their axons into the tectum opticum, but not into glial somata containing retina layers. We addressed the question whether the different glia of retina and tectum opticum differentially affect axon growth. Glial cells were purified from retina and tectum opticum by complement-mediated cytolysis of non-glial cells. RGCs were purified by enzymatic delayering from flat mounted retina. RGCs were seeded onto retinal versus tectal glia monolayers. Subsequent neuritic differentiation was analysed by immunofluorescence microscopy and scanning electron microscopy. Qualitative and quantitative evaluation revealed that retinal glia somata inhibited axons. Time-lapse video recording indicated that axonal inhibition was based on the collapse of lamellipodia- and filopodia-rich growth cones of axons. In contrast to retinal glia, tectal glia supported axonal extension. Notably, retinal glia were not inhibitory for neurons in general, because in control experiments axon extension of dorsal root ganglia was not hampered. Therefore, the axon inhibition by retinal glia was neuron type-specific. In summary, the data demonstrate that homotopic (retinal) glia somata inhibit axonal outgrowth of RGCs, whereas heterotopic (tectal) glia of the synaptic target area support RGC axon extension. The data underscore the pivotal role of glia in structuring the developing nervous system.     

Effects of capture and video‐recording on the behavior and breeding success of Great Tits in urban and forest habitats

Behavioral research often involves capturing and video‐recording birds, but these procedures may have undesired effects on the behavior of birds that have rarely been quantified. In addition, birds in urban and more natural areas may differ in their sensitivity to disturbance. We examined the possible effects of both capturing, weighing and measuring, and taking a blood sample, and the presence of video‐cameras on the behavior of male and female Great Tits (Parus major) breeding in urban and forest habitats. Using a 2 × 2 block design, we compared the behavior and breeding success of parents that either were or were not captured on their nests a few days before behavioral observations, and of parents that either were or were not habituated to the presence of a concealed video‐recorder mounted on nest boxes. We found no significant effects of habituation to the camera on bird behavior, but males captured in their nest boxes were more vigilant and hesitated longer before entering nest boxes, and also had slightly lower provisioning rates than males that had not been captured. Captured females also tended to be more vigilant than females that had not been captured, but their provisioning rates were not affected. Capturing males also influenced the behavior of their non‐captured mates, but capturing females had no effect on the behavior of their non‐captured mates. We found no difference in the effects of capture on Great Tits in urban and forest habitats, and our treatments also had no effect on the mass, size, and survival of nestlings until fledging. Our results suggest that, for Great Tits, being captured results in sex‐dependent behavioral effects that can last for at least several days. As such, we suggest that the possibility of similar effects in other species of birds should be considered in behavioral studies where birds must be captured, and recommend either that behavioral data be collected before capturing birds or that all birds in a study should be captured and handled in a standardized way.     

BEMOVI,software for extracting behavior and morphology from videos,illustrated with analyses of microbes

Microbes are critical components of ecosystems and provide vital services (e.g., photosynthesis, decomposition, nutrient recycling). From the diverse roles microbes play in natural ecosystems, high levels of functional diversity result. Quantifying this diversity is challenging, because it is weakly associated with morphological differentiation. In addition, the small size of microbes hinders morphological and behavioral measurements at the individual level, as well as interactions between individuals. Advances in microbial community genetics and genomics, flow cytometry and digital analysis of still images are promising approaches. They miss out, however, on a very important aspect of populations and communities: the behavior of individuals. Video analysis complements these methods by providing in addition to abundance and trait measurements, detailed behavioral information, capturing dynamic processes such as movement, and hence has the potential to describe the interactions between individuals. We introduce BEMOVI, a package using the R and ImageJ software, to extract abundance, morphology, and movement data for tens to thousands of individuals in a video. Through a set of functions BEMOVI identifies individuals present in a video, reconstructs their movement trajectories through space and time, and merges this information into a single database. BEMOVI is a modular set of functions, which can be customized to allow for peculiarities of the videos to be analyzed, in terms of organisms features (e.g., morphology or movement) and how they can be distinguished from the background. We illustrate the validity and accuracy of the method with an example on experimental multispecies communities of aquatic protists. We show high correspondence between manual and automatic counts and illustrate how simultaneous time series of abundance, morphology, and behavior are obtained from BEMOVI. We further demonstrate how the trait data can be used with machine learning to automatically classify individuals into species and that information on movement behavior improves the predictive ability.     

fMRI Validation of fNIRS Measurements During a Naturalistic Task

We present a method to compare brain activity recorded with near-infrared spectroscopy (fNIRS) in a dance video game task to that recorded in a reduced version of the task using fMRI (functional magnetic resonance imaging). Recently, it has been shown that fNIRS can accurately record functional brain activities equivalent to those concurrently recorded with functional magnetic resonance imaging for classic psychophysical tasks and simple finger tapping paradigms. However, an often quoted benefit of fNIRS is that the technique allows for studying neural mechanisms of complex, naturalistic behaviors that are not possible using the constrained environment of fMRI. Our goal was to extend the findings of previous studies that have shown high correlation between concurrently recorded fNIRS and fMRI signals to compare neural recordings obtained in fMRI procedures to those separately obtained in naturalistic fNIRS experiments. Specifically, we developed a modified version of the dance video game Dance Dance Revolution (DDR) to be compatible with both fMRI and fNIRS imaging procedures. In this methodology we explain the modifications to the software and hardware for compatibility with each technique as well as the scanning and calibration procedures used to obtain representative results. The results of the study show a task-related increase in oxyhemoglobin in both modalities and demonstrate that it is possible to replicate the findings of fMRI using fNIRS in a naturalistic task. This technique represents a methodology to compare fMRI imaging paradigms which utilize a reduced-world environment to fNIRS in closer approximation to naturalistic, full-body activities and behaviors. Further development of this technique may apply to neurodegenerative diseases, such as Parkinson’s disease, late states of dementia, or those with magnetic susceptibility which are contraindicated for fMRI scanning.     

Habitual action video game playing is associated with caudate nucleus-dependent navigational strategies

The habitual playing of video games is associated with increased grey matter and activity in the striatum. Studies in humans and rodents have shown an inverse relationship between grey matter in the striatum and hippocampus. We investigated whether action video game playing is also associated with increased use of response learning strategies during navigation, known to be dependent on the caudate nucleus of the striatum, when presented in a dual solution task. We tested 26 action video game players (actionVGPs) and 33 non-action video game players (nonVGPs) on the 4-on-8 virtual maze and a visual attention event-related potential (ERP) task, which elicits a robust N-2-posterior-controlateral (N2pc) component. We found that actionVGPs had a significantly higher likelihood of using a response learning strategy (80.76%) compared to nonVGPs (42.42%). Consistent with previous evidence, actionVGPs and nonVGPs differed in the way they deployed visual attention to central and peripheral targets as observed in the elicited N2pc component during an ERP visual attention task. Increased use of the response strategy in actionVGPs is consistent with previously observed increases in striatal volume in video game players (VGPs). Using response strategies is associated with decreased grey matter in the hippocampus. Previous studies have shown that decreased volume in the hippocampus precedes the onset of many neurological and psychiatric disorders. If actionVGPs have lower grey matter in the hippocampus, as response learners normally do, then these individuals could be at increased risk of developing neurological and psychiatric disorders during their lifetime.     

The ability of video image analysis to predict lean meat yield and EUROP score of lamb carcasses

A total of 862 lamb carcasses that were evaluated by both the VIAscan® and the current EUROP classification system were deboned and the actual yield was measured. Models were derived for predicting lean meat yield of the legs (Leg%), loin (Loin%) and shoulder (Shldr%) using the best VIAscan® variables selected by stepwise regression analysis of a calibration data set (n=603). The equations were tested on validation data set (n=259). The results showed that the VIAscan® predicted lean meat yield in the leg, loin and shoulder with an R² of 0.60, 0.31 and 0.47, respectively, whereas the current EUROP system predicted lean yield with an R² of 0.57, 0.32 and 0.37, respectively, for the three carcass parts. The VIAscan® also predicted the EUROP score of the trial carcasses, using a model derived from an earlier trial. The EUROP classification from VIAscan® and the current system were compared for their ability to explain the variation in lean yield of the whole carcass (LMY%) and trimmed fat (FAT%). The predicted EUROP scores from the VIAscan® explained 36% of the variation in LMY% and 60% of the variation in FAT%, compared with the current EUROP system that explained 49% and 72%, respectively. The EUROP classification obtained by the VIAscan® was tested against a panel of three expert classifiers (n=696). The VIAscan® classification agreed with 82% of conformation and 73% of the fat classes assigned by a panel of expert classifiers. It was concluded that VIAscan® provides a technology that can directly predict LMY% of lamb carcasses with more accuracy than the current EUROP classification system. The VIAscan® is also capable of classifying lamb carcasses into EUROP classes with an accuracy that fulfils minimum demands for the Icelandic sheep industry. Although the VIAscan® prediction of the Loin% is low, it is comparable to the current EUROP system, and should not hinder the adoption of the technology to estimate the yield of Icelandic lambs as it delivered a more accurate prediction for the Leg%, Shldr% and overall LMY% with negligible prediction bias.     

The production of female sex pheromone in Bactrocera oleae (Rossi) young males does not influence their mating chances

In Bactrocera oleae females attract males, which is in contrast to the majority of Tephritidae. However, the major component of the secretion of the female rectal ampulla glands, 1,7‐dioxaspiro‐[5,5]‐undecane (DSU), was also isolated from the glands of young males. The DSU produced by females and young males attracts males, but not females. In this study, we investigated the role of the production of DSU in young males. The mating performances of young and old males were evaluated, as well as the male‐male courtships by old males oriented at young and old males. Young males were found not to have a mating advantage. Young males were courted more by other males. Frame‐by‐frame analysis of male wing vibrations showed that this behavior did not differ when oriented at females and young males, highlighting that young males are perceived as females by the courting males. Overall, the production of DSU in young B. oleae males did not seem to be a case of female chemical mimicry. The hypothesis that young olive fruit fly males could benefit indirectly from the DSU production, simply distracting mature males away from females, is discussed.