Ringraziamo Joachim Erber per l&#39;introduzione di noi al paradigma dell&#39;apprendimento tattile di api mellifere. Questo lavoro è stato sostenuto dal cluster di 277 CITEC Excellence, finanziato nell&#39;ambito dell&#39;iniziativa eccellenza tedesca.

1. Preparazione delle api <ol><li> Bottinatrici nettare o polline sono catturati in campo sia da un alimentatore di saccarosio o direttamente dall&#39;ingresso alveare mentre tornava da un viaggio foraggiamento. Ogni singola ape viene catturata in una fiala di vetro che viene chiuso con un tappo di schiuma e presi immediatamente in laboratorio per ulteriori manipolazioni. </li><li> In laboratorio, le api catturate sono brevemente raffreddata in frigorifero a 4 ° C fino a quando i primi segni di immobilità. </li><li> Ogni singola ape immobilizzato è montato in un tubo metallico piccola con nastro adesivo tra la testa e il torace e sopra l&#39;addome. Si deve prestare attenzione che la proboscide e le antenne sono liberamente mobili. </li><li> Dipingere gli occhi composti e ocelli delle api fisso con vernice bianca (ad esempio, esente da solventi, Tipp-Ex) da occludere visione. </li><li> Aggiungere una goccia di cera fusa dietro la testa delle api per fissarlo al nastro tra la testa e il torace per evitare movimenti della testa durante le registrazioni. </li><li> Contrassegnare ogni singola ape con un numero sul nastro per una migliore identificazione e posizionare il tubo con l&#39;ape fisso in un ambiente umido per prevenire la disidratazione. </li><li> Alimentare ogni singola ape per 5 secondi con gocce di una soluzione di saccarosio al 30% presentato con una siringa e lasciare tutte le api recuperare per 30 min prima di iniziare con il protocollo condizionata tattile. </li></ol> 2. Condizionamento tattile <ol><li> Prima del condizionamento, ogni singola ape deve essere testato per la risposta estensione proboscide (PER) a uno stimolo di saccarosio 30% applicata alle antenne. In tal modo la punta della proboscide deve attraversare una linea virtuale tra le mandibole aperte. Eliminare tutte le api che non rispondono con un PER allo stimolo saccarosio. </li><li> Per il condizionamento tattile usare un cubo ottone (es. 3 x 5 mm) con una superficie liscia o un motivo inciso, ad esempio, scanalature orizzontali o verticali che formano un reticolo con 150 micron lunghezza d&#39;onda, come stimolo condizionato (CS). Fo lo stimolo incondizionato (US) utilizzare una soluzione di saccarosio al 30% (zucchero domestico, diluito in acqua). </li><li> Il cubo ottone (CS) è collocato in un supporto su un micromanipolatore (ad esempio, Märzhäuser MM33) per garantire l&#39;esatto posizionamento durante la procedura di condizionamento. L&#39;US è presentato al ape con una siringa riempita con una soluzione di saccarosio al 30%. </li><li> Il procedimento di condizionamento costituito da cinque coppie di stimolo tattile (CS) e soluzione di saccarosio (US), con un inter-trial intervallo (ITI) di 5 min. Posizionare una singola ape davanti al micromanipolatore con lo stimolo montato tattile (CS). Posizionare il CS lentamente, in modo tale che la superficie dello stimolo tattile è parallelo alla testa delle api (Figura 1 A e B). La distanza tra l&#39;animale e l&#39;oggetto tattile dovrebbe essere nell&#39;intervallo del raggio antennale lavoro dell&#39;ape testati, cioè l&#39;ape dovrebbe essere in grado di eseguire la scansione dello stimolo tattile in una posizione comoda wesimo entrambe le antenne. Nell&#39;esempio illustrato in figura 1, la distanza di 3 mm. Lasciate che l&#39;ape scansione lo stimolo tattile (CS) per 5 sec. Dopo i primi 3 sec, presentano una gocciolina di saccarosio 30% (US) soluzione con una siringa sotto la proboscide. Usare la punta della siringa per sollevare delicatamente la proboscide. Stimolazione Saccarosio sotto le proboscide susciterà l&#39;incondizionato PER 15. Lasciare l&#39;ape a leccare la ricompensa di saccarosio. Utilizzare un cronometro con un segnale di allarme per mantenere gli intervalli di tempo durante il condizionamento esatti. </li><li> Il PER condizionato viene usato come misura per il successo di apprendimento di un ape. Dopo gli accoppiamenti primo premiati, le api iniziano a rispondere alla presentazione CS per estensione dei loro proboscide, che indica che si aspettano la ricompensa futura. A proboscide massima estensione osservati in qualsiasi momento durante la finestra 3 sec Tempo di presentazione dello stimolo tattile e prima della presentazione saccarosio è segnato come una risposta positiva. Nessuna risposta è negativa contato. Il occurrence della PER Va osservato dallo sperimentatore. La percentuale di api che mostrano il PER in occasione della presentazione CS è tracciata per ogni prova. </li></ol> 3. Registrazioni cinematici <ol><li> Movimenti antennale di un&#39;ape sfruttata sono registrati con una videocamera digitale con un obiettivo macro adatto (per esempio Basler A602f-2 dotato di TechSpec VZM 200, operato a 50 fps tramite una connessione FireWire). La telecamera è posizionata sopra l&#39;animale in una vista dall&#39;alto in basso (Figura 1 A). In figura 1, la risoluzione spaziale delle registrazioni è di 0,02 mm / pixel. </li><li> Calibrare la fotocamera con la registrazione di immagini singole di una scacchiera 10 x 10 mm, con una lunghezza dello spigolo 1 mm da diversi orientamenti nel quadro dell&#39;obiettivo della fotocamera. La calibrazione può essere fatto con la cassetta degli attrezzi di calibrazione fotocamera Matlab 16. </li><li> Posizionare un un&#39;ape fissa sotto la lente della fotocamera. Presentare lo stimolo tattile, fissato ad un micromanipolatore, Per l&#39;animale. Procedere nello stesso modo descritto per il condizionamento tattile, e registrare il movimento antennale mentre l&#39;ape viene scansione dell&#39;oggetto. È importante che l&#39;intera gamma antennale lavoro e lo stimolo tattile sono visibili. Le scelte dei modelli di stimolo testato e numeri di prova dipende dal disegno sperimentale. </li></ol> 4. Analisi dei dati <ol><li> Il calcolo dello sfondo dell&#39;immagine è fatto in Matlab. Primo, il valore mediano di grigio nel tempo deve essere calcolato per ciascun pixel. Oggetti statici, come la testa fissa delle api e lo stimolo tattile, costituirà l&#39;immagine di sfondo. Oggetti in movimento, come due antenne, non sarà parte dello sfondo. </li><li> Ogni fotogramma del video registrato deve essere caricato in sequenza, e la differenza tra il frame corrente e lo sfondo dell&#39;immagine deve essere calcolata. Il risultato di questa sottrazione enfatizza quelle parti dell&#39;immagine che si sposta da frame a frame. Ideally, le antenne delle api sono le uniche aree di valori diversi da zero (Figura 1C). </li><li> Per l&#39;ulteriore elaborazione, i due più grandi aree con valori diversi da zero pixel vengono considerati le antenne. Per entrambe le antenne di una maschera binaria deve essere generato contenenti, per ciascun pixel un valore 1, se il pixel appartiene alla antenna e valore 0 altrimenti. Questa maschera serve come base per localizzare le punte antennali tardi. Per ottenere una maschera preliminare per l&#39;intera immagine del grigio-valore di ciascun pixel dell&#39;immagine differenza viene confrontata con una soglia predefinita. Dal momento che ci prendiamo cura di rumore entro la soglia viene scelto per essere piuttosto basso. La maschera preliminare poi porta ancora due tipi di errori: in primo luogo, a causa del rumore di immagine piccole regioni sono ancora parte della maschera. In secondo luogo, le aree che appartengono alla antenna potrebbe non necessariamente essere completamente collegato. Quest&#39;ultimo si verifica soprattutto se lo sfondo ha approssimativamente la stessa luminescenza come antenna. Per eliminare questi artefatti, standardoperazione di trattamento immagini morfologiche vengono applicate, ossia una combinazione di erosione e dilatazione dell&#39;immagine, basandosi sulle funzioni standard Matlab imerode e imdilate rispettivamente (vedi 17 pp 158-205 per ulteriori spiegazioni). Dopo il processo di denoising le ipotesi antenne sono ancora contenute all&#39;interno della maschera stessa. Quindi, come passo successivo la maschera binaria è in cluster per le aree disgiunte che utilizzano lo standard Matlab funzione bwlabel (vedi 17 pp 40-48 per ulteriori dettagli su l&#39;algoritmo di segmentazione di immagini). </li><li> Il numero di pixel per cluster viene contato e le due maggiori gruppi sono selezionati. Il centro di gravità è calcolata a distinguere tra l&#39;antenna sinistra e destra (Figura 1D). </li><li> La punta antennale per ciascuna antenna può essere definita come il pixel in un cluster con il valore massimo nella prossimale-distale direzione (vedi Figura 1D). </li></ol>

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Neurosci. 4, 46 (2010).</li><li>Scheiner, R., Barnert, M., Erber, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+in+water+and+sucrose+responsiveness+during+the+foraging+season+affects+proboscis+extension+learning+in+honey+bees.">Variation in water and sucrose responsiveness during the foraging season affects proboscis extension learning in honey bees.</a> Apidologie. 34, 67-72 (2003).</li><li>Mujagic, S., Erber, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sucrose+acceptance,+discrimination+and+proboscis+responses+of+honey+bees+(Apis+mellifera+L.)+in+the+field+and+the+laboratory.">Sucrose acceptance, discrimination and proboscis responses of honey bees (Apis mellifera L.) in the field and the laboratory.</a> J. Comp. Physiol. 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A. 181, 355-365 (1997).</li><li>Lambin, M., D&#233;glise, P., Gauthier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antennal+movements+as+indicators+of+odor+detection+by+worker+honeybees.">Antennal movements as indicators of odor detection by worker honeybees.</a> Apidologie. 36, 119-126 (2005).</li></ol>

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Preparazione delle api: raccogliere e fissare le api dovrebbe essere fatto rapidamente, al fine di mantenere il livello di stress di bassa animale. Lo stress ha un effetto sulla PER-risposta e, quindi, potrebbe avere un effetto indiretto sulle prestazioni di apprendimento delle api 19, 20. Il livello di stress può essere ridotta collocando i flaconi di vetro con le api direttamente su ghiaccio immediatamente dopo il prelievo per immobilizzare rapidamente. Si deve tenere conto che le api biso...

<table border="1"><tbody><tr><td> Nome del reagente / materiale </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti </td></tr><tr><td> Bottinatrici di polline, Apis mellifera L. </td><td></td><td></td><td> Orticaria privati </td></tr><tr><td> Fiale di vetro (22 x 45 mm) </td><td> Carl Roth GmbH &amp; Co </td><td> X655.1 </td><td></td></tr><tr><td> Tubi in ottone, 10 x 30 mm </td><td></td><td></td><td> Auto-prodotto </td></tr><tr><td> Ottone cubo, 3 x 5 mm </td><td></td><td></td><td> Auto-prodotto </td></tr><tr><td> Tipp-Ex (senza solventi) </td><td> Tipp-Ex GmbH &amp; Co KG </td><td></td><td></td></tr><tr><td> Soluzione di saccarosio (30%) </td><td></td><td></td><td> Suger domestica diluito in acqua </td></tr><tr><td> Etanolo (70%) </td><td> Carl Roth GmbH &amp; Co </td><td> 9.065,3 &lt;/ Td&gt; <td></td></tr><tr><td> Siringa (1 ml) e l&#39;ago (0,6 x 25) </td><td> Carl Roth GmbH &amp; Co </td><td> 59,1 </td><td></td></tr><tr><td> Stop Watch </td><td> Carl Roth GmbH &amp; Co </td><td> L423.1 </td><td></td></tr><tr><td> Micromanipolatore </td><td> Märzhäuser MM33 </td><td> 00-42-101-0000 </td><td></td></tr><tr><td> Videocamera digitale </td><td> Basler A602f-2 </td><td></td><td></td></tr><tr><td> Obiettivo Macro per fotocamera </td><td> TechSpec VZM 200 </td><td></td><td></td></tr><tr><td> Matlab R2009b </td><td> The MathWorks </td><td></td><td></td></tr></tbody></table>

tactile conditioning and movement analysis of antennal sampling strategies in honey bees (apis mellifera l.)

Le api da miele (Apis mellifera L.) sono insetti eusociali e ben noti per la loro complessa divisione del lavoro e la capacità di apprendimento associativo 1, 2. Le api operaie trascorrere la prima metà della loro vita all&#39;interno dell&#39;alveare buio, dove sono le larve allattamento o la costruzione della regolare pettini esagonale per il cibo (ad esempio polline o nettare) e la covata 3. Le antenne sono straordinari antenne multisensoriali e svolgono un ruolo fondamentale in diversi compiti tattili mediate 4, tra cui lo sviluppo alveare 5 e riconoscimento di pattern 6. Più tardi nella vita, ogni singola ape lascia l&#39;alveare alla ricerca di cibo. Poi un ape deve imparare a discriminare redditizie fonti di cibo, memorizzare la loro posizione, e comunicare ai propri compagni di nido 7. Api utilizzano diversi segnali floreali come colori o odori 7, 8, ma anche tattili dalla superficie petalo 9 per formare memorie multisensoriali della fonte di cibo. In laboratorio conditions, le api possono essere addestrati in un paradigma di apprendimento appetitivo di discriminare le caratteristiche degli oggetti tattili, come i bordi o scanalature con le loro antenne 10, 11, 12, 13. Questo paradigma di apprendimento è strettamente legato al condizionamento classico olfattivo della risposta estensione proboscide (PER) in api bardati 14. Il vantaggio del paradigma dell&#39;apprendimento tattile in laboratorio è la possibilità di combinare esperimenti comportamentali sull&#39;apprendimento con varie misurazioni fisiologiche, compresa l&#39;analisi del modello di movimento antennale.

Nel seguente esperimento abbiamo studiato come l&#39;apprendimento tattile influenza il comportamento antennale campionamento di api mellifere. Per questo, abbiamo monitorato il movimento della punta antennale in api pretrattati e condizionati prima e durante la presentazione di uno stimolo tattile. Primo, il movimento spontaneo antennale di un gruppo di raccoglitori polline (N = 42) è stata registrata per 1 min. Metà delle api (N = 21) è stata poi condizionata accoppiando cinque volte un...

Watch this Scientific Journal Video about Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L. at JoVE.com

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

In questo protocollo si mostra come condizione sfruttata api a stimoli tattili e introdurre un 2D tecnica del motion capture per analizzare la cinematica di fine scala modello di campionamento antenne.

Condizionata tattile e Analisi del Movimento di strategie di campionamento antennale di api mellifere (<em&gt; Apis mellifera</em&gt; L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

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Neuroscience

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

10.5K Views.  Bielefeld University. In questo protocollo si mostra come condizione sfruttata api a stimoli tattili e introdurre un 2D tecnica del motion capture per analizzare la cinematica di fine scala modello di campionamento antenne.

Video: Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

Eye Movement Monitoring of Memory

Explicit (often verbal) reports are typically used to investigate memory (e.g. "Tell me what you remember about the person you saw at the bank yesterday."), however such reports can often be unreliable or sensitive to response bias 1, and may be unobtainable in some participant populations. Furthermore, explicit reports only reveal when information has reached consciousness and cannot comment on when memories were accessed during processing, regardless of whether the information is subsequently accessed in a conscious manner. Eye movement monitoring (eye tracking) provides a tool by which memory can be probed without asking participants to comment on the contents of their memories, and access of such memories can be revealed on-line 2,3. Video-based eye trackers (either head-mounted or remote) use a system of cameras and infrared markers to examine the pupil and corneal reflection in each eye as the participant views a display monitor. For head-mounted eye trackers, infrared markers are also used to determine head position to allow for head movement and more precise localization of eye position. Here, we demonstrate the use of a head-mounted eye tracking system to investigate memory performance in neurologically-intact and neurologically-impaired adults. Eye movement monitoring procedures begin with the placement of the eye tracker on the participant, and setup of the head and eye cameras. Calibration and validation procedures are conducted to ensure accuracy of eye position recording. Real-time recordings of X,Y-coordinate positions on the display monitor are then converted and used to describe periods of time in which the eye is static (i.e. fixations) versus in motion (i.e., saccades). Fixations and saccades are time-locked with respect to the onset/offset of a visual display or another external event (e.g. button press). Experimental manipulations are constructed to examine how and when patterns of fixations and saccades are altered through different types of prior experience. The influence of memory is revealed in the extent to which scanning patterns to new images differ from scanning patterns to images that have been previously studied 2, 4-5. Memory can also be interrogated for its specificity; for instance, eye movement patterns that differ between an identical and an altered version of a previously studied image reveal the storage of the altered detail in memory 2-3, 6-8. These indices of memory can be compared across participant populations, thereby providing a powerful tool by which to examine the organization of memory in healthy individuals, and the specific changes that occur to memory with neurological insult or decline 2-3, 8-10.

Eye movement monitoring (or eye tracking) reveals where in space the eyes linger, when and for how long. Here, we demonstrate how eye tracking can be used to investigate the integrity of memory in multiple participant populations, without requiring verbal, or otherwise explicit, reports.

Eye Movement Monitoraggio della Memoria

Explicit (often verbal) reports are typically used to investigate memory (e.g. "Tell me what you remember about the person you saw at the bank ...

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Neuroscienze

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the survival of a honeybee colony depends on the exploitation of food sources, forager bees learn and memorize variable flower sites as well as their profitability. Forager bees can be easily trained in natural settings where they forage at a feeding site and learn the related signals such as odor or color. Appetitive associative learning can also be studied under controlled conditions in the laboratory by conditioning the proboscis extension response (PER) of individually harnessed honeybees3,4. This learning paradigm enables the study of the neuronal and molecular mechanisms that underlie learning and memory formation in a simple and highly reliable way5-12. A behavioral pharmacology approach is used to study molecular mechanisms. Drugs are injected systemically to interfere with the function of specific molecules during or after learning and memory formation13-16.
Here we demonstrate how to train harnessed honeybees in PER conditioning and how to apply drugs systemically by injection into the bee flight muscle.

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees Apis mellifera

We demonstrate how to implement a behavioral pharmacology method in an appetitive olfactory conditioning paradigm in honeybees (Apis mellifera) by systemic application of drugs. This method allows investigation of the mechanisms underlying learning and memory formation in a simple and reliable way.

Farmacologia Comportamentale nel condizionamento classico della risposta estensione proboscide in api ( Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the ...

Automated Sholl Analysis of Digitized Neuronal Morphology at Multiple Scales

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3. Specifically, it affects the ability of neurons to receive inputs from other cells2 and contributes to the propagation of action potentials4,5. The morphology of the neurites also affects how information is processed. The diversity of dendrite morphologies facilitate local and long range signaling and allow individual neurons or groups of neurons to carry out specialized functions within the neuronal network6,7. Alterations in dendrite morphology, including fragmentation of dendrites and changes in branching patterns, have been observed in a number of disease states, including Alzheimer's disease8, schizophrenia9,10, and mental retardation11. The ability to both understand the factors that shape dendrite morphologies and to identify changes in dendrite morphologies is essential in the understanding of nervous system function and dysfunction.
Neurite morphology is often analyzed by Sholl analysis and by counting the number of neurites and the number of branch tips. This analysis is generally applied to dendrites, but it can also be applied to axons. Performing this analysis by hand is both time consuming and inevitably introduces variability due to experimenter bias and inconsistency. The Bonfire program is a semi-automated approach to the analysis of dendrite and axon morphology that builds upon available open-source morphological analysis tools. Our program enables the detection of local changes in dendrite and axon branching behaviors by performing Sholl analysis on subregions of the neuritic arbor. For example, Sholl analysis is performed on both the neuron as a whole as well as on each subset of processes (primary, secondary, terminal, root, etc.) Dendrite and axon patterning is influenced by a number of intracellular and extracellular factors, many acting locally. Thus, the resulting arbor morphology is a result of specific processes acting on specific neurites, making it necessary to perform morphological analysis on a smaller scale in order to observe these local variations12.
The Bonfire program requires the use of two open-source analysis tools, the NeuronJ plugin to ImageJ and NeuronStudio. Neurons are traced in ImageJ, and NeuronStudio is used to define the connectivity between neurites. Bonfire contains a number of custom scripts written in MATLAB (MathWorks) that are used to convert the data into the appropriate format for further analysis, check for user errors, and ultimately perform Sholl analysis. Finally, data are exported into Excel for statistical analysis. A flow chart of the Bonfire program is shown in Figure 1.

We have developed a computer program to analyze neuronal morphology. In combination with two existing open source analysis tools, our program performs Sholl analysis and determines the number of neurites, branch points, and neurite tips. The analyses are performed so that local changes in neurite morphology can be observed.

Analisi automatizzata Sholl di Digitized morfologia neuronale a scale multiple

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3.  Specifically, it affects the ability of neurons to ...

Coherence between Brain Cortical Function and Neurocognitive Performance during Changed Gravity Conditions

Previous studies of cognitive, mental and/or motor processes during short-, medium- and long-term weightlessness have only been descriptive in nature, and focused on psychological aspects. Until now, objective observation of neurophysiological parameters has not been carried out - undoubtedly because the technical and methodological means have not been available -, investigations into the neurophysiological effects of weightlessness are in their infancy (Schneider et al. 2008).
While imaging techniques such as positron emission tomography (PET) and magnetic resonance imaging (MRI) would be hardly applicable in space, the non-invasive near-infrared spectroscopy (NIRS) technique represents a method of mapping hemodynamic processes in the brain in real time that is both relatively inexpensive and that can be employed even under extreme conditions. The combination with electroencephalography (EEG) opens up the possibility of following the electrocortical processes under changing gravity conditions with a finer temporal resolution as well as with deeper localization, for instance with electrotomography (LORETA).
Previous studies showed an increase of beta frequency activity under normal gravity conditions and a decrease under weightlessness conditions during a parabolic flight (Schneider et al. 2008a+b). Tilt studies revealed different changes in brain function, which let suggest, that changes in parabolic flight might reflect emotional processes rather than hemodynamic changes. However, it is still unclear whether these are effects of changed gravity or hemodynamic changes within the brain. Combining EEG/LORETA and NIRS should for the first time make it possible to map the effect of weightlessness and reduced gravity on both hemodynamic and electrophysiological processes in the brain. Initially, this is to be done as part of a feasibility study during a parabolic flight. Afterwards, it is also planned to use both techniques during medium- and long-term space flight.
It can be assumed that the long-term redistribution of the blood volume and the associated increase in the supply of oxygen to the brain will lead to changes in the central nervous system that are also responsible for anaemic processes, and which can in turn reduce performance (De Santo et al. 2005), which means that they could be crucial for the success and safety of a mission (Genik et al. 2005, Ellis 2000).
Depending on these results, it will be necessary to develop and employ extensive countermeasures. Initial results for the MARS500 study suggest that, in addition to their significance in the context of the cardiovascular and locomotor systems, sport and physical activity can play a part in improving neurocognitive parameters. Before this can be fully established, however, it seems necessary to learn more about the influence of changing gravity conditions on neurophysiological processes and associated neurocognitive impairment.

The effect of weightlessness and hypergravity on both hemodynamic and electrophysiological processes in the brain is going to be followed during parabolic flight by EEG and NIRS techniques. 
A feasibility study of a more complex experiment, which is planned to carry out during medium- and long-term space flight.

Coerenza tra funzione cerebrale corticale e prestazioni neurocognitive durante condizioni di gravità Cambiato

Previous studies of cognitive, mental and/or motor processes during short-, medium- and long-term weightlessness have only been descriptive in nature, and ...

Electrophysiological Measurements and Analysis of Nociception in Human Infants

Pain is an unpleasant sensory and emotional experience. Since infants cannot verbally report their experiences, current methods of pain assessment are based on behavioural and physiological body reactions, such as crying, body movements or changes in facial expression. While these measures demonstrate that infants mount a response following noxious stimulation, they are limited: they are based on activation of subcortical somatic and autonomic motor pathways that may not be reliably linked to central sensory processing in the brain. Knowledge of how the central nervous system responds to noxious events could provide an insight to how nociceptive information and pain is processed in newborns.
The heel lancing procedure used to extract blood from hospitalised infants offers a unique opportunity to study pain in infancy. In this video we describe how electroencephalography (EEG) and electromyography (EMG) time-locked to this procedure can be used to investigate nociceptive activity in the brain and spinal cord.
This integrative approach to the measurement of infant pain has the potential to pave the way for an effective and sensitive clinical measurement tool.

The assessment and treatment of pain in infants is difficult because infants cannot verbally report their experience. In this video we describe quantitative electrophysiological methods and analysis techniques that can be used to measure the response to noxious events from the infant nervous system.

Le misurazioni elettrofisiologiche e Analisi di nocicezione nei neonati umani

Pain is an unpleasant sensory and emotional experience. Since infants cannot verbally report their experiences, current methods of pain assessment are ...

Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity to normally innocuous stimuli1. Hyperalgesia and allodynia have been studied in many different rodent models of diabetes mellitus2. However, as stated by B&ouml;lcskei et al, determination of "pain" in animal models is challenging due to its subjective nature3. Moreover, the traditional methods used to determine behavioral responses to noxious thermal stimuli usually lack reproducibility and pharmacological sensitivity3. For instance, by using the hot-plate method of Ankier4, flinch, withdrawal and/or licking of either hind- and/or fore-paws is quantified as reflex latencies at constant high thermal stimuli (52-55 &deg;C). However, animals that are hyperalgesic to thermal stimulus do not reproducibly show differences in reflex latencies using those supra-threshold temperatures3,5. As the recently described method of B&ouml;lcskei et al.6, the procedures described here allows for the rapid, sensitive and reproducible determination of thermal nociceptive thresholds (TNTs) in mice and rats. The method uses slowly increasing thermal stimulus applied mostly to the skin of mouse/rat plantar surface. The method is particularly sensitive to study anti-nociception during hyperalgesic states such as PDN. The procedures described bellow are based on the ones published in detail by Alm&aacute;si et al 5 and B&ouml;lcskei et al 3. The procedures described here have been approved the Laboratory Animal Care and Use Committee (LACUC), Wright State University.

Here, we describe a rapid reliable and simple procedure to determine the lowest temperature at which rats or mice show nocifensive behavior, i.e. the thermal nociceptive threshold (TNT). This method applies a slowly increasing thermal stimulus allowing precise and reproducible estimation of TNTs with minimum, if any, stress to the animals.

Rapida determinazione della soglia termica nocicettivo in ratti diabetici

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity ...

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)

This video demonstrates novel techniques of RNA interference (RNAi) which downregulate two genes simultaneously in honey bees using double-stranded RNA (dsRNA) injections. It also presents a protocol of proboscis extension response (PER) assay for measuring gustatory perception.	
	RNAi-mediated gene knockdown is an effective technique downregulating target gene expression. This technique is usually used for single gene manipulation, but it has limitations to detect interactions and joint effects between genes. In the first part of this video, we present two strategies to simultaneously knock down two genes (called double gene knockdown). We show both strategies are able to effectively suppress two genes, vitellogenin (vg) and ultraspiracle (usp), which are in a regulatory feedback loop. This double gene knockdown approach can be used to dissect interrelationships between genes and can be readily applied in different insect species.	
	The second part of this video is a demonstration of proboscis extension response (PER) assay in honey bees after the treatment of double gene knockdown. The PER assay is a standard test for measuring gustatory perception in honey bees, which is a key predictor for how fast a honey bee's behavioral maturation is. Greater gustatory perception of nest bees indicates increased behavioral development which is often associated with an earlier age at onset of foraging and foraging specialization in pollen. In addition, PER assay can be applied to identify metabolic states of satiation or hunger in honey bees. Finally, PER assay combined with pairing different odor stimuli for conditioning the bees is also widely used for learning and memory studies in honey bees.

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees Apis mellifera

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

RNAi mediato doppia Gene atterramento e gustative Percezione misura di api mellifere (<em&gt; Apis mellifera</em&gt;)

This video demonstrates novel techniques of RNA  interference (RNAi) which downregulate two genes simultaneously in honey bees  using double-stranded RNA ...

Non-invasive Assessment of Changes in Corticomotoneuronal Transmission in Humans

The corticospinal pathway is the major pathway connecting the brain with the muscles and is therefore highly relevant for movement control and motor learning. There exists a number of noninvasive electrophysiological methods investigating the excitability and plasticity of this pathway. However, most methods are based on quantification of compound potentials and neglect that the corticospinal pathway consists of many different connections that are more or less direct. Here, we present a method that allows testing excitability of different fractions of the corticospinal transmission. This so called H-reflex conditioning technique allows one to assess excitability of the fastest (monosynaptic) and also polysynaptic corticospinal pathways. Furthermore, by using two different stimulation sites, the motor cortex and the cervicomedullary junction, it allows not only differentiation between cortical and spinal effects but also assessment of transmission at the corticomotoneural synapse. In this manuscript, we describe how this method can be used to assess corticomotoneural transmission after low-frequency repetitive transcranial magnetic stimulation, a method that was previously shown to reduce excitability of cortical cells. Here we demonstrate that not only cortical cells are affected by this repetitive stimulation but also transmission at the corticomotoneuronal synapse at the spinal level. This finding is important for the understanding of basic mechanisms and sites of neuroplasticity. Besides investigation of basic mechanisms, the H-reflex conditioning technique may be applied to test changes in corticospinal transmission following behavioral (e.g., training) or therapeutic interventions, pathology or aging and therefore allows a better understanding of neural processes that underlie movement control and motor learning.

The aim of the present study was to assess changes in transmission at the corticomotoneuronal synapses in humans after repetitive transcranial magnetic stimulation. For this purpose, an electrophysiological method is introduced that allows assessment of pathway specific corticospinal transmission, i.e. differentiation of fast, direct corticospinal pathways from polysynaptic connections.

Valutazione non invasiva dei cambiamenti nella trasmissione corticomoturonale negli esseri umani

The corticospinal pathway is the major pathway connecting the brain with the muscles and is therefore highly relevant for movement control and motor ...

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

Honey bees demonstrate astonishing learning abilities and advanced social behavior and communication. In addition, their brain is small, easy to visualize and to study. Therefore, bees have long been a favored model amongst neurobiologists and neuroethologists for studying the neural basis of social and natural behavior. It is important, however, that the experimental techniques used to study bees do not interfere with the behaviors being studied. Because of this, it has been necessary to develop a range of techniques for pharmacological manipulation of honey bees. In this paper we demonstrate methods for treating restrained or free-flying honey bees with a wide range of pharmacological agents. These include both noninvasive methods such as oral and topical treatments, as well as more invasive methods that allow for precise drug delivery in either systemic or localized fashion. Finally, we discuss the advantages and disadvantages of each method and&#160;describe common hurdles and how to best overcome them. We conclude with a discussion on the importance of adapting the experimental method to the biological questions rather than the other way around.

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

This manuscript describes several protocols for administering pharmacological agents to honey bees, including simple noninvasive methods for free-flying bees, as well as more invasive variants that allow precise localized treatment of restrained bees.

Manipolazione neurofarmacologico di sobrio e-volo libero api mellifere,<em&gt; Apis mellifera</em

Honey bees demonstrate astonishing learning abilities and advanced social behavior and communication. In addition, their brain is small, easy to visualize ...

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain undergoes rapid organizational change and is similar to accelerated brain changes that occur during the third trimester in humans. This model of fetal alcohol spectrum disorders (FASDs) produces severe brain damage, mimicking the amount and pattern of binge-drinking that occurs in some pregnant alcoholic mothers. We describe the use of trace eyeblink classical conditioning (ECC), a higher-order variant of associative learning, to assess long-term hippocampal dysfunction that is typically seen in alcohol-exposed adult offspring. At 90 days of age, rodents were surgically prepared with recording and stimulating electrodes, which measured electromyographic (EMG) blink activity from the left eyelid muscle and delivered mild shock posterior to the left eye, respectively. After a 5 day recovery period, they underwent 6 sessions of trace ECC to determine associative learning differences between alcohol-exposed and control rats. Trace ECC is one of many possible ECC procedures that can be easily modified using the same equipment and software, so that different neural systems can be assessed. ECC procedures in general, can be used as diagnostic tools for detecting neural pathology in different brain systems and different conditions that insult the brain.

Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings.

L'uso di traccia Eyeblink condizionamento classico per valutare la disfunzione Hippocampal in un modello del ratto dei disturbi dello spettro fetale alcol

Neonatal rats were administered a relatively high concentration of ethyl alcohol (11.9% v/v) during postnatal days 4-9, a time when the fetal brain ...

Condizionata tattile e Analisi del Movimento di strategie di campionamento antennale di api mellifere (

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