This work was supported by a grant from the Swiss National Science Foundation (CE: PBEZP3-131154)

1. Subject Recruitment
<ol>
<li>Subjects are recruited via an on-line database5 containing a list of about 3000 student volunteers. From this list we only contact women who have not previously participated in similar studies and ask them whether they are interested in participating in this study. We invite exclusively women as the time course of neurophysiological effects after acute testosterone administration is well established among women6,7. For men, this time course is not yet very well understood.</li>
<li>The interested subjects are then screened in a telephone interview to exclude medication intake, somatic diseases, hormonal contraception or any neurological or psychiatric disorders.</li>
<li>Because endogenous levels of sex hormones tend to be low and stable within the first part of the female menstrual cycle we conduct a second telephone interview to gain information about stability and duration of the cycle. Subjects are then invited to come to the experiment within 10 days from the beginning of their menstrual cycle. They are further instructed to abstain from alcohol or caffeine intake and smoking for 24 hours before the experiment.</li>
<li>Subjects are invited in groups of an average size of 12 to the experimental laboratory of the Institute for Empirical Research in Economics, in Zurich, Switzerland.</li>
</ol>
2. Experimental Procedure
<ol>
<li>All sessions start at 1.00 pm to control for the circadian variation in steroid hormone levels.</li>
<li>We conduct a pregnancy test before the beginning of the experiment to preclude pregnancy.</li>
<li>In preparation of the experiment, the testosterone and visually indistinguishable placebo pills are randomly distributed to twelve workstations marked one to twelve within the laboratory room by a person who is not involved in the experiment otherwise.</li>
<li>At the entrance to the laboratory, each of the twelve subject draws a card with a concealed number on it. The numbers range from one to twelve.</li>
<li>Subjects are then taken to the laboratory room which is endowed with a total of 36 computer workplaces. Subjects are instructed to seek out and sit down by the workplace marked with the same number as indicated on the card they drew at the entrance.</li>
<li>All subjects read and sign the informed consent form.</li>
<li>We measure baseline salivary testosterone and cortisol concentrations using the IBL SaliCaps Kit (IBL-Hamburg, Hamburg, Germany) for saliva specimen collection. Subjects are requested to transfer 1 mL of saliva into the test tubes, which are then immediately frozen at -80&deg;C.</li>
<li>Subjects are requested to take the sublingual pill into their mouth, to chew it and keep the disintegrated substance in their mouth for one minute. To achieve maximal contact of the HP-&beta;-CD testosterone inclusion complex with the oral mucosa, subjects are instructed to make circling movements with their tongue. After one minute, the remainder of the substance is swallowed using 100 mL water.</li>
<li>Immediately following the substance administration, subjects are required to complete different questionnaires. We measure mood and arousal by means of the Multidimensional Mood Questionnaire (MDBF)8 and anxiety and anger traits and states, by means of the State-Trait Anxiety Inventory9 and the State-Trait Anger Expression Inventory10. These questionnaires have the purpose of controlling for any possible effects of subjects' incidental mood, anger and anxiety states as well as for personality characteristics. Furthermore, subjects complete the validated German versions of the 90-item Symptom Checklist (revised version)11 to measure their psychological health. Those who have very high scores (two standard deviations above the mean, indicating poor psychological health) are excluded from the data analysis. 
Due to the established time lag of four hours for the appearance of behavioral effects after sublingual application of 0.5 mg testosterone in young, healthy women6,7, a long waiting period is an integral part of our study. During this period, our subjects are required to stay in the laboratory room and keep themselves quietly occupied. This is to ensure that no social interaction outside the laboratory takes place which could give rise to confounded measures.</li>
</ol>
3. Ultimatum Game
<ol>
<li>Subjects then receive the instructions for their role in the game and the payoff rules on paper and are asked to read them. We check their comprehension of the payoff structure by having them complete several control questions at the end of the instructions.</li>
<li>We then summarize the experimental procedure orally. The reason for this is to ensure that subjects know that everyone else has the same information about the structure of the game and how the payoffs are determined, which is also referred to as "common knowledge". The summary is as follows:</li>
<li>"The Ultimatum Game is played with two types of players, a proposer and a responder, who have to agree on the division of 10 monetary units (MUs). The proposer can decide on the distribution of the 10 MUs. In this experiment, the proposers can propose offers of 5, 3, 2, or 0 MUs to the responder. The responder then has to either accept or reject this proposal. If the responder accepts the proposer's offer, the proposed allocation will be implemented and paid. However, the responder can also reject the proposal; in this case neither party earns anything." </li>
<li>We ensure a clear separation between an offer that is regarded as fair (5 MUs) and offers that are perceived as unfair (3, 2 or 0 MUs) by restricting the offers to 5, 3, 2 or 0 MUs. Subjects are randomly and anonymously assigned to the role of either the proposer or the responder and then remain in this role for the whole experiment. Every proposer makes three propositions on the distribution of MUs while paired with three different randomly selected interaction partners. No pair of subjects interacts more than once. They never learn the identity of the persons with whom they are matched in the experiment. The proposer does not receive feedback about the responders' choices until the end of the experiment. All decisions in the ultimatum game are implemented in zTree software and presented on computer screens12.</li>
<li>Because testosterone is one of the most widely discussed hormones in the popular press it is possible that subjects have a preformed belief (true or false) about its effects13. Furthermore, because ethical concerns require researchers to inform subjects that they will either receive a placebo or testosterone the prior knowledge about testosterone may pollute a possible impact of testosterone on behavior. Therefore we assess the perceived group assignment by asking the subjects whether they think they have received testosterone or the placebo pill.</li>
<li>The perceived group assignment is recorded in the final questionnaire together with a few socioeconomic characteristics. During the last questionnaire we prepare subjects' payment.</li>
</ol>
4. Subject Payment
<ol>
<li>Subjects receive a base fee of one hundred Swiss francs for participation in the experiment. The use of real monetary incentives greatly reduces a potential social desirability bias and biased self-perceptions. Therefore, each subjects receives one Swiss franc (approximately 1 US$) for each MU earned in the ultimatum game. Each subject receives his or her earnings consisting of the base fee plus the earned MUs in private at the end of the experiment.</li>
</ol>

<ol>
	<li>Edwards, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+Androgen+Stimulation+and+Aggressive+Behavior+in+Male+and+Female+Mice.">Early Androgen Stimulation and Aggressive Behavior in Male and Female Mice.</a> Physiology &#38; Behav. 4, 333-338 (1969).</li><li>Archer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testosterone+and+human+aggression:+an+evaluation+of+the+challenge+hypothesis.">Testosterone and human aggression: an evaluation of the challenge hypothesis.</a> Neurosci Biobehav R. 30, 319-345 (2006).</li><li>Mazur, A., Booth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testosterone+and+dominance+in+men.">Testosterone and dominance in men.</a> Behav Brain Sci. 21, 353-363 (1998).</li><li>Josephs, R. A., Newman, M. L., Brown, R. P., Beer, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Status,+testosterone,+and+human+intellectual+performance:+stereotype+threat+as+status+concern.">Status, testosterone, and human intellectual performance: stereotype threat as status concern.</a> Psychol Sci. 14, 158-163 (2003).</li><li>Greiner, B., Kremer, K., Macho, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Forschung und wissenschaftliches Rechnen: GWDG Bericht 63. , 79-93 (2004).</li><li>van Honk, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+single+administration+of+testosterone+induces+cardiac+accelerative+responses+to+angry+faces+in+healthy+young+women.">A single administration of testosterone induces cardiac accelerative responses to angry faces in healthy young women.</a> Behav Neurosci. 115, 238-242 (2001).</li><li>van Honk, J., Peper, J. S., Schutter, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testosterone+reduces+unconscious+fear+but+not+consciously+experienced+anxiety:+implications+for+the+disorders+of+fear+and+anxiety.">Testosterone reduces unconscious fear but not consciously experienced anxiety: implications for the disorders of fear and anxiety.</a> Biol Psychiatry. 58, 218-225 (2005).</li><li>Steyer, R., Schwenkmezger, P., Notz, P., Eid, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Der Mehrdimensionale Befindlichkeitsfragebogen (MDBF) [Multidimensional Mood Questionnaire. , (1997).</li><li>Laux, L., Glanzmann, P., Schaffner, P., Spielberger, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Das State-Trait-Angstinventar (Manual for the State-Trait Anxiety Inventory. , (1981).</li><li>Schwenkmezger, P., Hodapp, V., Spielberger, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Das State-Trait-Aergerausdruck-Inventar STAXI [The State-Trait Anger Expression Inventory (STAXI)]. , (1992).</li><li>Derogatis, L. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> SCL-90-R, administration, scoring &amp;amp; procedures manual-I for the R(evised) version. , (1977).</li><li>Fischbacher, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=z-Tree:+Zurich+toolbox+for+ready-made+economic+experiments.">z-Tree: Zurich toolbox for ready-made economic experiments.</a> Experimental Economics. 10, 171-178 (2007).</li><li>Bjorkqvist, K., Nygren, T., Bjorklund, A. C., Bjorkqvist, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Testosterone+Intake+and+Aggressiveness+-+Real+Effect+or+Anticipation.">Testosterone Intake and Aggressiveness - Real Effect or Anticipation.</a> Aggressive Behav. 20, 17-26 (1994).</li><li>Camerer, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Behavioral game theory: experiments in strategic interaction. , (2003).</li><li>Henrich, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term="Economic+man"+in+cross-cultural+perspective:+Behavioral+experiments+in+15+small-scale+societies.">"Economic man" in cross-cultural perspective: Behavioral experiments in 15 small-scale societies.</a> Behavioral and Brain Sciences. 28, 795-855 (2005).</li><li>Bardsley, N., Cubitt, R. P., Loomes, G., Moffatt, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Experimental economics : rethinking the rules. , (2009).</li><li>Eisenegger, C., Naef, M., Snozzi, R., Heinrichs, M., Fehr, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prejudice+and+truth+about+the+effect+of+testosterone+on+human+bargaining+behaviour.">Prejudice and truth about the effect of testosterone on human bargaining behaviour.</a> Nature. 463, 356-359 (2010).</li></ol>

No conflicts of interest declared.

An indication that your results are valid is if the average offers observed is similar to what other studies found. Typically, average offers are between 30 - 40% of the total amount and approximately half of the subjects offer the equal share14. On the other hand, a likely indicator that subjects did not understand the rules of the game is if offers of more than 50% are observed. It is worth noting, however, that there is considerable heterogeneity in the distribution and average offers observed in studies ar...

combining behavioral endocrinology and experimental economics: testosterone and social decision making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

Watch this Scientific Journal Video about Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making at JoVE.com

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

combining-behavioral-endocrinology-experimental-economics

Research

Education

JoVE Core

JoVE Journal

Introduction to Psychology

Neuroscience

Unit 1

Biopsychology

SCIENTISTS IN ACTION

15.0K Views.  University of Zurich. The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

Video: Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Combining Computer Game-Based Behavioural Experiments With High-Density EEG and Infrared Gaze Tracking

Experimental paradigms are valuable insofar as the timing and other parameters of their stimuli are well specified and controlled, and insofar as they yield data relevant to the cognitive processing that occurs under ecologically valid conditions. These two goals often are at odds, since well controlled stimuli often are too repetitive to sustain subjects' motivation. Studies employing electroencephalography (EEG) are often especially sensitive to this dilemma between ecological validity and experimental control: attaining sufficient signal-to-noise in physiological averages demands large numbers of repeated trials within lengthy recording sessions, limiting the subject pool to individuals with the ability and patience to perform a set task over and over again. This constraint severely limits researchers' ability to investigate younger populations as well as clinical populations associated with heightened anxiety or attentional abnormalities. Even adult, non-clinical subjects may not be able to achieve their typical levels of performance or cognitive engagement: an unmotivated subject for whom an experimental task is little more than a chore is not the same, behaviourally, cognitively, or neurally, as a subject who is intrinsically motivated and engaged with the task. A growing body of literature demonstrates that embedding experiments within video games may provide a way between the horns of this dilemma between experimental control and ecological validity. The narrative of a game provides a more realistic context in which tasks occur, enhancing their ecological validity (Chaytor &amp; Schmitter-Edgecombe, 2003). Moreover, this context provides motivation to complete tasks. In our game, subjects perform various missions to collect resources, fend off pirates, intercept communications or facilitate diplomatic relations. In so doing, they also perform an array of cognitive tasks, including a Posner attention-shifting paradigm (Posner, 1980), a go/no-go test of motor inhibition, a psychophysical motion coherence threshold task, the Embedded Figures Test (Witkin, 1950, 1954) and a theory-of-mind (Wimmer &amp; Perner, 1983) task. The game software automatically registers game stimuli and subjects' actions and responses in a log file, and sends event codes to synchronise with physiological data recorders. Thus the game can be combined with physiological measures such as EEG or fMRI, and with moment-to-moment tracking of gaze. Gaze tracking can verify subjects' compliance with behavioural tasks (e.g. fixation) and overt attention to experimental stimuli, and also physiological arousal as reflected in pupil dilation (Bradley et al., 2008). At great enough sampling frequencies, gaze tracking may also help assess covert attention as reflected in microsaccades - eye movements that are too small to foveate a new object, but are as rapid in onset and have the same relationship between angular distance and peak velocity as do saccades that traverse greater distances. The distribution of directions of microsaccades correlates with the (otherwise) covert direction of attention (Hafed &amp; Clark, 2002).

Procedures for recording high-density EEG and gaze data during computer game-based cognitive tasks are described. Using a video game to present cognitive tasks enhances ecological validity without sacrificing experimental control.

Experimental paradigms are valuable insofar as the timing and other parameters of their stimuli are well specified and controlled, and insofar as they ...

Brain Imaging Investigation of the Neural Correlates of Observing Virtual Social Interactions

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language affect our decisions in personal and professional life alike 1. These "friend or foe" judgements are often based on first impressions, which in turn may affect our decisions to "approach or avoid". Previous studies investigating the neural correlates of social cognition tended to use static facial stimuli 2. Here, we illustrate an experimental design in which whole-body animated characters were used in conjunction with functional magnetic resonance imaging (fMRI) recordings. Fifteen participants were presented with short movie-clips of guest-host interactions in a business setting, while fMRI data were recorded; at the end of each movie, participants also provided ratings of the host behaviour. This design mimics more closely real-life situations, and hence may contribute to better understanding of the neural mechanisms of social interactions in healthy behaviour, and to gaining insight into possible causes of deficits in social behaviour in such clinical conditions as social anxiety and autism 3.

This article demonstrates an experimental design in which whole-body animated characters are used in conjunction with functional magnetic resonance imaging (fMRI) to investigate the neural correlates of observing virtual social interactions.

The ability to gauge social interactions is crucial in the assessment of others&#x2019; intentions. Factors such as facial expressions and body language ...

Measuring the Subjective Value of Risky and Ambiguous Options using Experimental Economics and Functional MRI Methods

Most of the choices we make have uncertain consequences. In some cases the probabilities for different possible outcomes are precisely known, a condition termed "risky". In other cases when probabilities cannot be estimated, this is a condition described as "ambiguous". While most people are averse to both risk and ambiguity1,2, the degree of those aversions vary substantially across individuals, such that the subjective value of the same risky or ambiguous option can be very different for different individuals. We combine functional MRI (fMRI) with an experimental economics-based method3 to assess the neural representation of the subjective values of risky and ambiguous options4. This technique can be now used to study these neural representations in different populations, such as different age groups and different patient populations.
In our experiment, subjects make consequential choices between two alternatives while their neural activation is tracked using fMRI. On each trial subjects choose between lotteries that vary in their monetary amount and in either the probability of winning that amount or the ambiguity level associated with winning. Our parametric design allows us to use each individual's choice behavior to estimate their attitudes towards risk and ambiguity, and thus to estimate the subjective values that each option held for them. Another important feature of the design is that the outcome of the chosen lottery is not revealed during the experiment, so that no learning can take place, and thus the ambiguous options remain ambiguous and risk attitudes are stable. Instead, at the end of the scanning session one or few trials are randomly selected and played for real money. Since subjects do not know beforehand which trials will be selected, they must treat each and every trial as if it and it alone was the one trial on which they will be paid. This design ensures that we can estimate the true subjective value of each option to each subject. We then look for areas in the brain whose activation is correlated with the subjective value of risky options and for areas whose activation is correlated with the subjective value of ambiguous options.

Using functional MRI and behavioral methods to determine the neural representation of the subjective value of risky and ambiguous options in the human brain.

Most of the choices we make have uncertain consequences. In some cases the probabilities for different possible outcomes are precisely known, a condition ...

C. elegans Tracking and Behavioral Measurement

We have developed instrumentation, image processing, and data analysis techniques to quantify the locomotory behavior of C. elegans as it crawls on the surface of an agar plate. For the study of the genetic, biochemical, and neuronal basis of behavior, C. elegans is an ideal organism because it is genetically tractable, amenable to microscopy, and shows a number of complex behaviors, including taxis, learning, and social interaction1,2. Behavioral analysis based on tracking the movements of worms as they crawl on agar plates have been particularly useful in the study of sensory behavior3, locomotion4, and general mutational phenotyping5. Our system works by moving the camera and illumination system as the worms crawls on a stationary agar plate, which ensures no mechanical stimulus is transmitted to the worm. Our tracking system is easy to use and includes a semi-automatic calibration feature. A challenge of all video tracking systems is that it generates an enormous amount of data that is intrinsically high dimensional. Our image processing and data analysis programs deal with this challenge by reducing the worms shape into a set of independent components, which comprehensively reconstruct the worms behavior as a function of only 3-4 dimensions6,7. As an example of the process we show that the worm enters and exits its reversal state in a phase specific manner.

C. elegans Tracking and Behavioral Measurement

We have developed a video-rate tracking microscope system that can record and quantify C. elegans behavior at high resolution and high speeds. We have also developed computational methods to reduce the dimensionality of the worm images to a fundamental set of measurements that completely describe the shape of the worm.

We have developed instrumentation, image processing, and data analysis techniques to quantify the locomotory behavior of C. elegans as it crawls on the ...

A Low Cost Setup for Behavioral Audiometry in Rodents

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the experiments. Such parameters may be physiological response characteristics of the auditory pathway, e.g. via brainstem audiometry (BERA). But these methods allow only indirect and uncertain extrapolations about the auditory percept that corresponds to these physiological parameters. To assess the perceptual level of hearing, behavioral methods have to be used. A potential problem with the use of behavioral methods for the description of perception in animal models is the fact that most of these methods involve some kind of learning paradigm before the subjects can be behaviorally tested, e.g. animals may have to learn to press a lever in response to a sound. As these learning paradigms change perception itself 1,2 they consequently will influence any result about perception obtained with these methods and therefore have to be interpreted with caution. Exceptions are paradigms that make use of reflex responses, because here no learning paradigms have to be carried out prior to perceptual testing. One such reflex response is the acoustic startle response (ASR) that can highly reproducibly be elicited with unexpected loud sounds in na&iuml;ve animals. This ASR in turn can be influenced by preceding sounds depending on the perceptibility of this preceding stimulus: Sounds well above hearing threshold will completely inhibit the amplitude of the ASR; sounds close to threshold will only slightly inhibit the ASR. This phenomenon is called pre-pulse inhibition (PPI) 3,4, and the amount of PPI on the ASR gradually depends on the perceptibility of the pre-pulse. PPI of the ASR is therefore well suited to determine behavioral audiograms in na&iuml;ve, non-trained animals, to determine hearing impairments or even to detect possible subjective tinnitus percepts in these animals. In this paper we demonstrate the use of this method in a rodent model (cf. also ref. 5), the Mongolian gerbil (Meriones unguiculatus), which is a well know model species for startle response research within the normal human hearing range (e.g. 6).

A fast and inexpensive method for the behavioral determination of hearing parameters like hearing thresholds, hearing impairments or phantom perceptions (subjective tinnitus) is described. It uses pre-pulse inhibition of the acoustic startle response and can be easily implemented in a personal computer using a programmable AD/DA-converter and a piezo sensor.

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance imaging. However, use of such methods in animal models requires anesthesia and hence both alters the brain state and prevents behavioral measures. An alternative method is the use of in vivo microdialysis to take continuous measurement of brain extracellular fluid concentrations of glucose, lactate, and related metabolites in awake, unrestrained animals. This technique is especially useful when combined with tasks designed to rely on specific brain regions and/or acute pharmacological manipulation; for example, hippocampal measurements during a spatial working memory task (spontaneous alternation) show a dip in extracellular glucose and rise in lactate that are suggestive of enhanced glycolysis1-3,4-5, and intrahippocampal insulin administration both improves memory and increases hippocampal glycolysis6. Substances such as insulin can be delivered to the hippocampus via the same microdialysis probe used to measure metabolites. The use of spontaneous alternation as a measure of hippocampal function is designed to avoid any confound from stressful motivators (e.g. footshock), restraint, or rewards (e.g. food), all of which can alter both task performance and metabolism; this task also provides a measure of motor activity that permits control for nonspecific effects of treatment. Combined, these methods permit direct measurement of the neurochemical and metabolic variables regulating behavior.

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Modulation of hippocampally-dependent spatial working memory by direct intrahippocampal microinjection, accompanied and followed by in vivo microdialysis for metabolites in conscious, behaving animals.

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific frequency and in turn modulate ongoing cortical oscillatory activity. This neurotool allows the establishment of a causal link between endogenous oscillatory activity and behavior. Most of the tACS studies have shown online effects of tACS. However, little is known about the underlying action mechanisms of this technique because of the AC-induced artifacts on Electroencephalography (EEG) signals. Here we show a unique approach to investigate online physiological frequency-specific effects of tACS of the primary motor cortex (M1) by using single pulse Transcranial Magnetic Stimulation (TMS) to probe cortical excitability changes. In our setup, the TMS coil is placed over the tACS electrode while Motor Evoked Potentials (MEPs) are collected to test the effects of the ongoing M1-tACS. So far, this approach has mainly been used to study the visual and motor systems. However, the current tACS-TMS setup can pave the way for future investigations of cognitive functions. Therefore, we provide a step-by-step manual and video guidelines for the procedure.

Transcranial Alternating Current Stimulation (tACS) allows the modulation of cortical excitability in a frequency-specific fashion. Here we show a unique approach which combines online tACS with single pulse Transcranial Magnetic Stimulation (TMS) in order to "probe" cortical excitability by means of Motor Evoked Potentials.

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...

Operant Protocols for Assessing the Cost-benefit Analysis During Reinforced Decision Making by Rodents

Reinforcement-guided decision making is the ability to choose between competing courses of action based on the relative value of the benefits and their consequences. This process is integral to the normal human behavior and has been shown to be disrupted by neurological and psychiatric disorders such as addiction, schizophrenia, and depression. Rodents have long been used to uncover the neurobiology of human cognition. To this end, several behavioral tasks have been developed; however, most are non-automated and are labor-intensive. The recent development of the open-source microcontroller has enabled researchers to automate operant-based tasks for assessing a variety of cognitive tasks, standardizing the stimulus presentation, improving the data recording and consequently, improving the research output. Here, we describe an automated delay-based reinforcement-guided decision-making task, using an operant T-maze controlled by custom-written software programs. Using these decision-making tasks, we show the changes in the local field potential activities in the anterior cingulate cortex of a rat whilst it performs a delay-based cost-and-benefit decision-making task.

A cost-benefit analysis is a weighing-scale approach that the brain performs during the course of decision making. Here, we propose a protocol to train rats on an operant-based decision-making paradigm where rats choose higher rewards at the expense of waiting for 15 s to receive them.

Reinforcement-guided decision making is the ability to choose between competing courses of action based on the relative value of the benefits and their ...

Abbiategrasso Brain Bank Protocol for Collecting, Processing and Characterizing Aging Brains

In a constantly aging population, the prevalence of neurodegenerative disorders is expected to rise. Understanding disease mechanisms is the key to find preventive and curative measures. The most effective way to achieve this is through direct examination of diseased and healthy brain tissue. The authors present a protocol to obtain, process, characterize and store good quality brain tissue donated by individuals registered in an antemortem brain donation program. The donation program includes a face-to-face empathic approach to people, a collection of complementary clinical, biological, social and lifestyle information and serial multi-dimensional assessments over time to track individual trajectories of normal aging and cognitive decline. Since many neurological diseases are asymmetrical, our brain bank offers a unique protocol for slicing fresh specimens. Brain sections of both hemispheres are alternately frozen (at -80 &#176;C) or fixed in formalin; a fixed slice on one hemisphere corresponds to a frozen one on the other hemisphere. With this approach, a complete histological characterization of all frozen material can be obtained, and omics studies can be performed on histologically well-defined tissues from both hemispheres thus offering a more complete assessment of neurodegenerative disease mechanisms. Correct and definite diagnosis of these diseases can only be achieved by combining the clinical syndrome with the neuropathological evaluation, which often adds important etiological clues necessary to interpret the pathogenesis. This method can be time consuming, expensive and limited as it only covers a limited geographical area. Regardless of its limitations, the high degree of characterization it provides can be rewarding. Our ultimate goal is to establish the first Italian Brain Bank, all the while emphasizing the importance of neuropathologically verified epidemiological studies.

This protocol describes a method to track individual brain-aging trajectories through a brain donation program and proper characterization of brains. Brain donors are involved in a long-term longitudinal study including serial multi-dimensional assessments. The protocol contains a detailed description of brain processing and an accurate diagnostic methodology.

In a constantly aging population, the prevalence of neurodegenerative disorders is expected to rise. Understanding disease mechanisms is the key to find ...

Post-Movie Subliminal Measurement (PMSM), for Investigating Implicit Social Bias

New knowledge is continuously gained from a social environment that can influence how people respond to each other. Such responses often occur implicitly, at a subliminal perceptual level, and related brain mechanisms can be experimentally isolated by presenting the stimuli quickly. Subliminal presentation of faces that belong to different ethnicity groups, races, or gender has been shown to be successful in investigating social implicit responses. However, many implicit responses are based on knowledge previously gained about the faces (e.g., sexual orientation, political views, and socioeconomic status) and not solely on physical appearance. Here, a novel method called post-movie subliminal measurement (PMSM) is presented. When watching a socially engaging movie, a spectator gains knowledge about the protagonist and becomes familiar with his/her identity and world views. When the face of the protagonist is presented subliminally after the movie, it evokes an implicit neural response depending on what is learned about the protagonist. With a vast number of movies available, each depicting a variety of people with different identities, the PMSM method enables investigation of the brain&#39;s complex implicit biases in a manner that resembles real-life social perceptions.

Post-Movie Subliminal Measurement PMSM, for Investigating Implicit Social Bias

This protocol describes the use of movies to investigate brain mechanisms underlying implicit social biases during functional magnetic resonance imaging. When the face of a protagonist is presented after a movie subliminally, it evokes an implicit response based on knowledge of the protagonist gained during the movie.

New knowledge is continuously gained from a social environment that can influence how people respond to each other. Such responses often occur implicitly, ...