Name: a method to study adaptation to left-right reversed audition
Uploaded: 2018-10-29T12:30:00
Description: Watch this Scientific Journal Video about A Method to Study Adaptation to Left-Right Reversed Audition at JoVE.com

This work was partially supported by a grant from JSPS KAKENHI Grant Number JP17K00209. The author thanks Takayuki Hoshino and Kazuhiro Shigeta for technical assistance.

All methods described here have been approved by the Ethics Committee of Tokyo Denki University. For every participant, informed consent was obtained after the participant received a detailed explanation of the protocol.
1. Setup of the Left-Right Reversed Audition System
<ol>
	<li>Setup of the Reversed Audition System without a Participant 
	<ol>
		<li>Prepare a linear pulse-code-modulation (LPCM) recorder, binaural microphones, and binaural in-ear earphones.</li>
		<li>Con...

<ol>
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The proposed protocol aimed to establish a methodology for studying adaptation to left-right reversed audition as an effective tool for uncovering the adaptability of humans to a novel auditory environment. As evidenced by the representative results, the constructed apparatus achieved left-right reversed audition with high spatiotemporal accuracy. Although the previous apparatuses for reversed audition11,12,13,<sup cl...

Adaptability to a novel environment is one of the fundamental functions for humans to live robustly in any situation. One effective tool for uncovering the mechanism of environmental adaptability in humans is an unusual sensory space that is artificially produced by apparatuses. In the majority of the previous studies dealing with this topic, special spectacles with prisms have been used to achieve left-right reversed vision1,2,3,4,5 or up-down reversed vision6,7. Furthermore, exposure to such vision from a few days to more than a month has revealed perceptual and behavioral adaptation1,2,3,4,5,6,7 (e.g., capability to ride a bicycle2,5,7). Moreover, periodic measurements of the brain activity using neuroimaging techniques, such as electroencephalography (EEG)1, magnetoencephalography (MEG)3, and functional magnetic resonance imaging (fMRI)2,4,5,7, have detected changes in the neural activity underlying the adaptation (e.g., bilateral visual activation for unilateral visual stimulation4,5). Although the participant&#39;s appearance becomes strange to some extent and great care is needed for the observer to maintain the participant&#39;s safety, reversed vision with prisms provides precise three-dimensional (3D) visual information without any delay in a wearable manner. Therefore, the methodology for uncovering the mechanism of environmental adaptability is relatively established in the visual domain.
In 1879, Thompson proposed a concept of pseudophone, &#34;an instrument for investigating the laws of binaural audition by means of the illusions it produces in the acoustic perception of space&#34;8. However, in contrast to the visual cases1,2,3,4,5,6,7, few attempts have been made to study the adaptation to unusual auditory spaces, and no noticeable knowledge has been obtained to date. Despite a long history of developing virtual auditory displays9,10, wearable apparatuses for controlling 3D audition have rarely been developed. Hence, only a few reports examined the adaptation to left-right reversed audition. One traditional apparatus consists of a pair of curved trumpets that are crossed and inserted into a participant&#39;s ear canals in a contrariwise manner11,12. In 1928, Young first reported the use of these crossed trumpets and wore them continuously for 3 days at most or a total of 85 h to test adaptation to left-right reversed audition. Willey et al.12 retested the adaptation in three participants wearing the trumpets for 3, 7, and 8 days, respectively. The curved trumpets easily provided left-right reversed audition, but had an issue with the reliability of spatial accuracy, wearability, and strange appearance. A more advanced apparatus for the reversed audition is an electronic system in which left and right lines of head/earphones and microphones are reversely connected13,14. Ohtsubo et al.13 achieved auditory reversal using the first ever binaural headphone-microphones that were connected to a fixed amplifier and evaluated its performance. More recently, Hofman et al.14 cross-linked complete-in-canal hearing aids and tested adaptation in two participants that wore the aids for 49 h in 3 days and 3 weeks, respectively. Although these studies have reported high performance of sound source localization in the front auditory field, the sound source localization in the backfield and a potential delay of electrical devices have never been evaluated. Especially in Hofman et al.&#39;s study, the spatial performance of the hearing aids was guaranteed for the front 60&#176; in the head-fixed condition and for the front 150&#176; in the head-free condition, suggesting unknown omniazimuth performance. Moreover, the exposure period may be too short to detect phenomena related to the adaptation as compared with the longer cases of reversed vision2,4,5. None of these studies have measured brain activity using neuroimaging techniques. Therefore, the uncertainty in spatiotemporal accuracy, the short exposure periods, and the non-utilization of neuroimaging could be reasons for the small number of reports and the limited amount of knowledge on adaptation to left-right reversed audition.
Thanks to the recent advances in wearable acoustic technology, Aoyama and Kuriki15 succeeded in constructing a left-right reversed 3D audition using only wearable devices that recently became available and achieved the omniazimuth system with high spatiotemporal accuracy. Moreover, approximately a 1-month exposure to reversed audition using the apparatus exhibited some representative results for MEG measurements. Based on this report, we describe, in this article, a detailed protocol to set-up, validate and use the system, and to test the adaptation to left-right reversed audition with the help of neuroimaging that is performed periodically without the system. This approach is effective for uncovering the adaptability of humans to a novel environment in the auditory domain.

<table>
	<tbody>
		<tr>
			<td>Linear pulse-code-modulation recorder</td>
			<td>Sony</td>
			<td>PCM-M10</td>
			<td></td>
		</tr>
		<tr>
			<td>Binaural microphones</td>
			<td>Roland</td>
			<td>CS-10EM</td>
			<td></td>
		</tr>
		<tr>
			<td>Binaural in-ear earphones</td>
			<td>Etymotic Research</td>
			<td>ER-4B</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital angle protractor</td>
			<td>Wenzhou Sanhe Measuring Instrument</td>
			<td>5422-200</td>
			<td></td>
		</tr>
		<tr>
			<td>Plane-wave speaker</td>
			<td>Alphagreen</td>
			<td>SS-2101</td>
			<td></td>
		</tr>
		<tr>
			<td>Video camera</td>
			<td>Sony</td>
			<td>HDR-CX560</td>
			<td></td>
		</tr>
		<tr>
			<td>MATLAB</td>
			<td>Mathworks</td>
			<td>R2012a, R2015a</td>
			<td>R2012a for stimulation and R2015a for analysis</td>
		</tr>
		<tr>
			<td>Psychophysics Toolbox</td>
			<td>Free</td>
			<td>Version 3</td>
			<td>http://psychtoolbox.org</td>
		</tr>
		<tr>
			<td>Insert earphones</td>
			<td>Etymotic Research</td>
			<td>ER-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetoencephalography system</td>
			<td>Neuromag</td>
			<td>Neuromag-122 TM</td>
			<td></td>
		</tr>
		<tr>
			<td>Electroencephalography system</td>
			<td>Brain Products</td>
			<td>acti64CHamp</td>
			<td></td>
		</tr>
		<tr>
			<td>MNE</td>
			<td>Free</td>
			<td>MNE Software Version 2.7, 
			MNE 0.13</td>
			<td>https://martinos.org/mne/stable/index.html</td>
		</tr>
		<tr>
			<td>The Multivariate Granger Causality Toolbox</td>
			<td>Free</td>
			<td>mvgc_v1.0</td>
			<td>http://www.sussex.ac.uk/sackler/mvgc/</td>
		</tr>
	</tbody>
</table>

a method to study adaptation to left-right reversed audition

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the previous studies have used special spectacles with prisms to achieve unusual spaces in the visual domain, a methodology for studying the adaptation to unusual auditory spaces has yet to be fully established. This study proposes a new protocol to set-up, validate, and use a left-right reversed stereophonic system using only wearable devices, and to study the adaptation to left-right reversed audition with the help of neuroimaging. Although individual acoustic characteristics are not yet implemented, and slight spillover of unreversed sounds is relatively uncontrollable, the constructed apparatus shows high performance in a 360&#176; sound source localization coupled with hearing characteristics with little delay. Moreover, it looks like a mobile music player and enables a participant to focus on daily life without arousing curiosity or drawing attention of other individuals. Since the effects of adaptation were successfully detected at the perceptual, behavioral, and neural levels, it is concluded that this protocol provides a promising methodology for studying adaptation to left-right reversed audition, and is an effective tool for uncovering the adaptability of humans to a novel environments in the auditory domain.

The representative results shown here are based on Aoyama and Kuriki15. The present protocol achieved left-right reversed audition with high spatiotemporal accuracy. Figure 1 shows the sound source localization in directions over 360&#176; before and immediately after putting on the left-right reversed audition system (Figure 1A), in six participants, as indicated by the cosine similarity. As shown in <str...

Watch this Scientific Journal Video about A Method to Study Adaptation to Left-Right Reversed Audition at JoVE.com

A Method to Study Adaptation to Left-Right Reversed Audition

The present study proposes a protocol to investigate the adaptation to left-right reversed audition achieved only by wearable devices, using neuroimaging, which can be an effective tool for uncovering the adaptability of humans to a novel environment in the auditory domain.

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the ...

This method can be an effective tool for uncovering the adaptability of humans to a novel environment in the auditory domain. The main advantage with this technique is that long-term adaptation to precise left-right reversed audition can be tested in a variable manner in combination with neural imaging. Demonstrating the procedure will be Takayuki Hoshino, a grad student from my laboratory.To begin, prepare the linear pulse code modulation recorder, binaural microphones and binaural in-ear earphones. First, cross-connect the left and right lines of the microphones to the recorder so that left-right reversed analog sound signals are digitalized. Then, connect the left and right lines of the earphones straight through to the recorder so that the reversed digitalized signals are played immediately.Finally, put the bodies of the microphones and the earphones together for each ear with slight isolation by soundproofing materials. Then cover the microphones with dedicated windscreens for suppressing any wind noise. Next, insert rechargeable batteries and a large-capacity, high-speed memory card into the recorder and turn it on.Then place the body of the system into a pocket-sized bag. Instruct the participant to insert the earphones tightly into the ear canals. Then disconnect the lines for the left and right microphones and connect the dominant ear side of the microphone straight through to the recorder.Now, have them repetitively take off and put on the dominant ear side of the system while adjusting the sound volume of the recorder. This makes the subjective loudness of the reversed sounds equal. Ah, ah.Check the loudness for the non-dominant ear, as well. Then reconnect all the system lines. Tell the participant that, during exposure, they should follow the same calibration procedure whenever they set up the system again.Prior to reversed audition exposure, have the participant thoroughly practice the task that will be used during the neuroimaging experiments. A selective reaction time task can be used. Use a Psychophysics software Toolbox to present 1000 Hertz sounds at 65 decibels sound pressure level for 0.1 seconds, played 80 times per block.Also use an interstimulus interval of 2.5 to 3.5 seconds, played pseudorandomly in either ear. The task should consist of two compatible and two incompatible blocks. In the compatible condition, the participant should respond immediately to the sound with the index finger on the same side of their body as the sound.This should be alternated with the incompatible condition in which the participant responds immediately to the sound with the index finger on the opposite side of their body. Before the exposure to reversed audition, conduct a neuroimaging experiment using the trained task. Record either MEG or EEG responses, as well as the left and right finger responses.To begin exposure, provide the participant with a sufficient number of spare rechargeable batteries and large-capacity, high-speed memory cards so they can replace them as needed. Then, remind them repeatedly of their right to quit the exposure at any time and instruct them to wear, calibrate and check the reversed audition system by themselves during the exposure period. The participant should perform daily life activities while wearing the system continuously for approximately a month, except while sleeping, bathing, neuroimaging or during an emergency.When removed, the system should be immediately replaced with earplugs without producing any sound while in a silent area. The batteries and memory cards should also be replaced routinely, before battery exhaustion and memory overcapacity. To facilitate adaptation, the participant should experience situations involving high auditory input, such as playing video games, walking in a shopping mall or a campus and having a conversation with more than two persons for as long as possible.They should also keep a diary or provide a subjective report to an observer as often as possible, detailing perceptual and behavioral changes, experienced events or any other details. During approximately a one month exposure to reversed audition, conduct neuroimaging experiments under the trained task every week without the reversed audition system in exactly the same way as in the preexposure experiment. Finally, after the target exposure period, the participant should take off the reversed audition system.One week after exposure had ended, conduct a final neuroimaging experiment under the trained task. This figure shows the sound source localization in directions over 360 degrees before and immediately after putting on the left-right reversed audition system in six participants. Here, cosine similarity between perceptual angles and sine-regulated physical angles in the normal and reversed conditions are plotted against unregulated physical angles.And here, we see cosine similarity between perceptual angles in the reversed condition and oppositely-arranged perceptual angles in the normal condition plotted against physical angles. This figure shows changes in behavioral and neural responses during the selective reaction time task before, during and after exposure in one participant. Yellow zones indicate a period exposed to left-right reversed audition.Mean reaction times for the stimulus response compatible and incompatible conditions are seen here, while here we see left and right auditory N1m intensities for stimulus response compatible and incompatible conditions as evaluated by minimum norm estimates of MEG data. This figure shows auditory motor functional connectivity as tested by Granger causality tests during the selective reaction time task in two participants. The left and right motor and auditory areas are shown.Red, yellow or lack of arrows indicate the number of participants who showed significance at a threshold of P less than 0.05. After watching this video, you should have a good understanding of how to study left-right reversed audition as a tool to uncover the adaptability of humans to a novel auditory environment.

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Behavior

Uncovering Beat Deafness: Detecting Rhythm Disorders with Synchronized Finger Tapping and Perceptual Timing Tasks

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here with the goal of uncovering rhythm disorders, such as beat deafness. Beat deafness is characterized by poor performance in perceiving durations in auditory rhythmic patterns or poor synchronization of movement with auditory rhythms (e.g., with musical beats). These tasks include the synchronization of finger tapping to the beat of simple and complex auditory stimuli and the detection of rhythmic irregularities (anisochrony detection task) embedded in the same stimuli. These tests, which are easy to administer, include an assessment of both perceptual and sensorimotor timing abilities under different conditions (e.g., beat rates and types of auditory material) and are based on the same auditory stimuli, ranging from a simple metronome to a complex musical excerpt. The analysis of synchronized tapping data is performed with circular statistics, which provide reliable measures of synchronization accuracy (e.g., the difference between the timing of the taps and the timing of the pacing stimuli) and consistency. Circular statistics on tapping data are particularly well-suited for detecting individual differences in the general population. Synchronized tapping and anisochrony detection are sensitive measures for identifying profiles of rhythm disorders and have been used with success to uncover cases of poor synchronization with spared perceptual timing. This systematic assessment of perceptual and sensorimotor timing can be extended to populations of patients with brain damage, neurodegenerative diseases (e.g., Parkinson&#8217;s disease), and developmental disorders (e.g., Attention Deficit Hyperactivity Disorder).

Behavioral tasks that allow for the assessment of perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) are presented. Synchronization of finger tapping to the beat of an auditory stimuli and detecting rhythmic irregularities provides a means of uncovering rhythm disorders.

A set of behavioral tasks for assessing perceptual and sensorimotor timing abilities in the general population (i.e., non-musicians) is presented here ...

Rodent Brain Microinjection to Study Molecular Substrates of Motivated Behavior

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as appropriate models, partly because the response to behaviorally relevant stimuli and the circuitry parsing stimulus-action outcomes is astonishingly similar between humans and rodents. In studying molecular substrates of complex behaviors, the microinjection of reagents that modify, augment, or silence specific systems is an invaluable technique. However, it is crucial that the microinjection site is precisely targeted in order to aid interpretation of the results. We present a method for the manufacture of surgical implements and microinjection needles that enables accurate microinjection and unlimited customizability with minimal cost. Importantly, this technique can be successfully completed in awake rodents if conducted in conjunction with other JoVE articles that covered requisite surgical procedures. Additionally, there are many behavioral paradigms that are well suited for measuring motivation. The progressive ratio is a commonly used method that quantifies the efficacy of a reinforcer to maintain responding despite an (often exponentially) increasing work requirement. This assay is sensitive to reinforcer magnitude and pharmacological manipulations, which allows reinforcing efficacy and/ or motivation to be determined. We also present a straightforward approach to program operant software to accommodate a progressive ratio reinforcement schedule.

Rodents are an appropriate model to investigate the molecular substrates of behavior and complex psychiatric disorders. Brain microinjection in awake rodents can be used to elucidate disease substrates. An efficient and customizable brain microinjection method as well as the execution of an operant paradigm that quantifies motivation is presented.

Brain microinjection can aid elucidation of the molecular substrates of complex behaviors, such as motivation. For this purpose rodents can serve as ...

Visualizing Visual Adaptation

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical or age differences, or a color deficiency or disease. This protocol describes a technique for incorporating sensory adaptation into the simulations. The protocol is illustrated with the example of color vision, but is generally applicable to any form of visual adaptation. The protocol uses a simple model of human color vision based on standard and plausible assumptions about the retinal and cortical mechanisms encoding color and how these adjust their sensitivity to both the average color and range of color in the prevailing stimulus. The gains of the mechanisms are adapted so that their mean response under one context is equated for a different context. The simulations help reveal the theoretical limits of adaptation and generate &#34;adapted images&#34; that are optimally matched to a specific environment or observer. They also provide a common metric for exploring the effects of adaptation within different observers or different environments. Characterizing visual perception and performance with these images provides a novel tool for studying the functions and consequences of long-term adaptation in vision or other sensory systems.

This article describes a novel method for simulating and studying adaptation in the visual system.

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical ...

Psychophysical Tracking Method to Measure Taste Preferences in Children and Adults

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to adulthood. The method, which is identical for children, adolescents, and adults, is of short duration (&#60; 15 min), does not rely on sustained attention or place demands on memory (which would yield spurious age differences), and minimizes the impact of language development, making this method amenable to the cognitive limitations of pediatric populations. In this whole-mouth tasting method, subjects are asked to taste (without swallowing) pairs of solutions of different sucrose concentrations and to point to the solution they prefer. Each subsequent pair contains the participant&#39;s preceding preferred concentration and an adjacent stimulus concentration. The procedure continues until the subject chooses either a given concentration of sucrose when paired with both a higher and a lower concentration, or the highest or lowest concentration two consecutive times. Subjects are prevented from reaching response criteria on the basis of first or second position bias by the two-series design of the method, which counterbalances the order of solution presentation within each pair between the series (the weaker concentration is presented first in Series 1, second in Series 2). The geometric mean of the two sucrose concentrations chosen in Series 1 and 2 is an estimate of the participant&#39;s most preferred level of sucrose. Sucrose preference as determined with this laboratory-based measure has been shown to be associated with preference for sugars in foods and beverages and with taste receptor genotype, family history of alcoholism, and race/ethnicity, as well as depressive symptomatology among pediatric populations. The method has real-world relevance and has been applied to determine most preferred level of other tastes (e.g., salt), making it a valuable psychophysical tool.

Sweet taste has powerful hedonic appeal among people of all ages, particularly children. Described herein is a reliable and valid method that can be used to determine the level of sweetness most preferred, making it a valuable psychophysical tool for scientists.

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to ...

Using a Split-belt Treadmill to Evaluate Generalization of Human Locomotor Adaptation

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. However, studying human locomotor learning can be challenging. During infancy and childhood, the neuromuscular system is quite immature, and it is unlikely that locomotor learning during early stages of development is governed by the same mechanisms as in adulthood. By the time humans reach maturity, they are so proficient at walking that it is difficult to come up with a sufficiently novel task to study de novo locomotor learning. The split-belt treadmill, which has two belts that can drive each leg at a different speed, enables the study of both short- (i.e., immediate) and long-term (i.e., over minutes-days; a form of motor learning) gait modifications in response to a novel change in the walking environment. Individuals can easily be screened for previous exposure to the split-belt treadmill, thus ensuring that all experimental participants have no (or equivalent) prior experience. This paper describes a typical split-belt treadmill adaptation protocol that incorporates testing methods to quantify locomotor learning and generalization of this learning to other walking contexts. A discussion of important considerations for designing split-belt treadmill experiments follows, including factors like treadmill belt speeds, rest breaks, and distractors. Additionally, potential but understudied confounding variables (e.g., arm movements, prior experience) are considered in the discussion.

We describe a protocol for investigating human locomotor adaptation using the split-belt treadmill, which has two belts that can drive each leg at a different speed. We specifically focus on a paradigm designed to test the generalization of adapted locomotor patterns to different walking contexts (e.g., gait speeds, walking environments).

Understanding the mechanisms underlying locomotor learning helps researchers and clinicians optimize gait retraining as part of motor rehabilitation. ...

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors remains poorly understood. In Drosophila melanogaster, environmental cues, such as food availability, affect mating activity offering a tractable system to investigate the mechanisms modulating sexual behavior. In D. melanogaster, environmental cues are often sensed via the chemosensory gustatory and olfactory systems. Here, we present a method to test the effect of environmental chemical cues on mating behavior. The assay consists of a small mating arena containing food medium and a mating couple. The mating frequency for each couple is continuously monitored for 24 h. Here we present the applicability of this assay to test environmental compounds from an external source through a pressurized air system as well as manipulation of the environmental components directly in the mating arena. The use of a pressurized air system is especially useful to test the effect of very volatile compounds, while manipulating components directly in the mating arena can be of value to ascertain a compound's presence. This assay can be adapted to answer questions about the influence of genetic and environmental cues on mating behavior and fecundity as well as other male and female reproductive behaviors.

A Method to Test the Effect of Environmental Cues on Mating Behavior in Drosophila melanogaster

We demonstrate an assay to analyze the environmental and genetic cues that influence mating behavior in the fruit fly Drosophila melanogaster.

An individual's sexual drive is influenced by genotype, experience and environmental conditions. How these factors interact to modulate sexual behaviors ...

A Computational Method to Quantify Fly Circadian Activity

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. Fundamental mechanisms that underlie this temporal control are widely studied using the fruit fly Drosophila melanogaster as a model organism. In flies, the clock is typically studied by analyzing multiday locomotor recording. Such a recording shows a complex bimodal pattern with two peaks of activity: a morning peak that happens around dawn, and an evening peak that happens around dusk. These two peaks together form a waveform that is very different from sinusoidal oscillations observed in clock genes, suggesting that mechanisms in addition to the clock have profound effects in producing the observed patterns in behavioral data. Here we provide instructions on using a recently developed computational method that mathematically describes temporal patterns in fly activity. The method fits activity data with a model waveform that consists of four exponential terms and nine independent parameters that fully describe the shape and size of the morning and evening peaks of activity. The extracted parameters can help elucidate the kinetic mechanisms of substrates that underlie the commonly observed bimodal activity patterns in fly locomotor rhythms.

A method to quantify the main temporal features seen in fly circadian locomotor rhythms is presented. The quantification is achieved by fitting fly activity with a multi-parametric model waveform. The model parameters describe the shape and size of the morning and evening peaks of daily activity.

In most animals and plants, circadian clocks orchestrate behavioral and molecular processes and synchronize them to the daily light-dark cycle. ...

Modified Blood Collection from Tail Veins of Non-anesthetized Mice with a Vacuum Blood Collection System and Eyeglass Magnifier

Blood sample collection is the basis of experimental animal research. It is of importance to obtain adequate blood samples for various research purposes. The tail veins of mice are small, and it is sometimes difficult to obtain the required blood volume by conventional puncture methods. This study investigates the superiority of repeated blood sample collection from tail veins of mice through use of a vacuum blood collection system and eyeglass magnifier (experimental group) compared to conventional blood sampling methods (conventional group), performed by beginners and experts, respectively. With the help of an eyeglass magnifier, a butterfly needle tip is inserted into the tail vein of each mouse in the experimental group. When the vein is penetrated successfully, a blood sample is collected in the vacuum collection tube by insertion of the rubber end of a butterfly needle into the vacuum blood collection tube. The plunger is then used to collect blood without the help of the eyeglass magnifier in the conventional group. Success rates of blood sample collection by the beginners and experts were shown to be 70% vs. 100% (p &#60; 0.01) in the experimental group and 35% vs. 85% (p &#60; 0.01) in the conventional group. For both beginners and experts, puncture times required for obtaining required blood sample were significantly lower in the experimental group compared to the conventional group (2.40 &#177; 0.75 vs. 2.90 &#177; 0.31, p &#60; 0.05; 1.15 &#177; 0.37 vs. 1.55 &#177; 0.76, p &#60; 0.05). In conclusion, the presented blood sampling technique is feasible and easy to practice and enables frequent sampling of adequate blood volumes from non-anesthetized mice.&#160;

This study reports blood sampling from tail vein in mice using a vacuum extraction tube system with eyeglass magnifier. Our method is easy to practice and could be used for repeat blood sampling in mice.

Blood sample collection is the basis of experimental animal research. It is of importance to obtain adequate blood samples for various research purposes. ...

A Laboratory Method to Measure Contagious Yawning in Rats

Communication is an essential aspect of animal social life. Animals may influence one another and come together in schools, flocks, and herds. Communication is also the way sexes interact during courtship and how rivals settle disputes without fighting. However, there are some behavioral patterns for which it is difficult to test the existence of a communicatory function, because several types of sensory modalities are likely involved. For example, contagious yawning is a communicatory act in mammals that potentially occurs through sight, hearing, smell, or a combination of these senses depending on whether the animals are familiar to one another. Therefore, to test hypotheses about the possible communicatory role of such behaviors, a suitable method is necessary to identify the participating sensory modalities.
The method proposed here aims to obtain yawn contagion curves for familiar and unfamiliar rats and evaluate the relative participation of visual and olfactory sensory modalities. The method uses inexpensive materials, and with some minor changes, it can also be used with other rodent species such as mice. Overall, the method involves the substitution of clear dividers (with or without holes) with opaque dividers (with or without holes) that either allow or prevent communication between rats placed in adjacent cages with holes in adjoining sides. Accordingly, four conditions can be tested: olfactory communication, visual communication, both visual and olfactory communication, and neither visual nor olfactory communication. As social interaction occurs between the rats, these test conditions simulate what may occur in a natural environment. In this respect, the method proposed here is more effective than traditional methods that rely on video presentations whose biological validity can raise concerns. Nonetheless, it does not discriminate between the potential role of hearing and roles of smell and vision in yawn contagion.

The method described here aims to obtain yawn contagion curves in pairs of familiar or unfamiliar male rats. Cages with holes separated by either clear or opaque partitions with (or without) holes are used to detect whether visual, olfactory, or both types of sensory cues can stimulate yawn contagion.

Communication is an essential aspect of animal social life. Animals may influence one another and come together in schools, flocks, and herds. ...

A Novel Pavlovian Fear Conditioning Paradigm to Study Freezing and Flight Behavior

Fear- and anxiety-related behaviors significantly contribute to an organism&#8217;s survival. However, exaggerated defensive responses to perceived threat are characteristic of various anxiety disorders, which are the most prevalent form of mental illness in the United States. Discovering the neurobiological mechanisms responsible for defensive behaviors will aid in the development of novel therapeutic interventions. Pavlovian fear conditioning is a widely used laboratory paradigm to study fear-related learning and memory. A major limitation of traditional Pavlovian fear conditioning paradigms is that freezing is the only defensive behavior monitored. We recently developed a modified Pavlovian fear conditioning paradigm that allows us to study both conditioned freezing and flight (also known as escape) behavior within individual subjects. This model employs higher intensity footshocks and a greater number of pairings between the conditioned stimulus and unconditioned stimulus. Additionally, this conditioned flight paradigm utilizes serial presentation of pure tone and white noise auditory stimuli as the conditioned stimulus. Following conditioning in this paradigm, mice exhibit freezing behavior in response to the tone stimulus, and flight responses during the white noise. This conditioning model can be applied to the study of rapid and flexible transitions between behavioral responses necessary for survival.

Defensive behavioral responses are contingent upon threat intensity, proximity, and context of exposure. Based on these factors, we developed a classical conditioning paradigm that elicits clear transitions between conditioned freezing and flight behavior within individual subjects. This model is crucial for the understanding the pathologies involved in anxiety, panic, and post-traumatic stress disorders.

Fear- and anxiety-related behaviors significantly contribute to an organism&#8217;s survival. However, exaggerated defensive responses to perceived threat ...