This work was supported by a Howard Hughes Medical Institute Education Grant and an internal grant from the Saint Joseph's University chapter of Sigma Xi. We would also like to thank the three anonymous reviewers who helped strengthen the protocol and interpretations.

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The authors (GW and SPM) have no competing financial interests or conflicts of interest.

הפרוטוקול יקבע אם יש עמותות עקביות תעוזה, תוקפנות, והתנהגויות shoaling דג הזברה. אם יש עמותות עקביות באוכלוסייה נתונה בין כל ההתנהגויות האלה, ואז תסמונת התנהגותית היא הווה. על ידי לימוד תסמונת התנהגותית הטבעי של האוכלוסייה, החוקרים יכולים לקבל הבנה מלאה יותר של דינמי שלה...

ישנו גוף הולך וגדל של ספרות חוקר את הקשר בין התנהגויות שונות בתוך חיות בודדות באוכלוסיית נתונה. עמותות אלה מכונות תסמונות התנהגותיות, והמדידות כוללות בדרך כלל תעוזה, תוקפנות, התנהגות גישוש, ו חברותיות 1-5. תסמונות התנהגותיות הן בעלי ערך עבור שני סיבות ישירות ועקיפות. ישירות, ידע של תסמונות התנהגותיות יכול לספק תמונה מלאה יותר של תורת האבולוציה, מבנה האוכלוסייה, ואת הדינמיקה באוכלוסייה 3. בעקיפין, ידע לגבי עמותות התנהגותי עשוי להודיע ​​שדות לכמת התנהגות כגון פרמקולוגיה 6, טוקסיקולוגיה 7, גנטיקה התנהגותית 8,9, ו אנדוקרינולוגיה 10. בגלל יתרונות ישירים ועקיפים אלה, ידע מוגבר תסמונות התנהגותיות הוא בעל ערך במיוחד אורגניזמים מודל נפוצים כמו דג הזברה. מחקרים שעשו שימוש דג הזברה נמצאים iדיסציפלינות נה, כולל ניתוח של תסמונות התנהגותיות 11-13. כדי לקדם את הידע במחקר תסמונת התנהגותית, ובגלל דיסציפלינות אחרות גם להשתמש במדידות התנהגותי בדיקת שערות, תיאורים אמינים ותמציתיים של התנהגות נדרשים עבור ניתוחים ופרשנויות תקפים ופרוטוקולים סטנדרטיים יקלו השוואות בין-מחקר בתוך מינים. פרוטוקול שלנו פותחה כדי למדוד תסמונת התנהגותית תעוזה-תוקפנות-shoaling בקרב אוכלוסיה של דג הזברה מעבדה שגדלו 14. עם זאת, הבסיס של הפרוטוקול (מעקב דגים בודדים, הבטחה אקראית ראוי, ניתוחים מתאימים) ניתן לשנות בקלות עבור מגוון רחב של אמצעי התנהגותיים אלטרנטיביים. בנוסף, התעוזה, תוקפנות, או מבחני shoaling ניתן להפעיל בנפרד עבור הבדיקות של השערות ברורות. לכן, בזמן שהוא המטרה שלנו היא לתאר כיצד לערוך מחקר תסמונת התנהגותית ופרוטוקול עבור l הפרט מוצלחמדידת איבל התנהגותי, כל היבט של הליך זה יכול לעמוד לבד. הספרות על תסמונות התנהגותיות חובקת מספר קבוצות טקסונומיות, מן פרוקי רגליים לבני אדם 4 ו, על מנת למדוד תסמונת התנהגותית, לפחות בשני קשרים ההתנהגותיים חייבים לכמת. למרבה הצער, יש לעתים קרובות עקבי מעט המבחנים המשמשים לכמת את המדידה התנהגותי פני הצירים של התנהגות. לדוגמא, בדגים, תעוזה ניתן למדוד באמצעות מבחני מבוך T, מבחני שדה פתוח, או הקדמה של רומן או גירוי חוץ 15. מחקרי תוקפנות בדגים עלולים להיות כרוכים אינטראקציות הדיאדה, מבחני גירוי וידאו, או מבחנים מחימר 12,16,17. כמו כן, ניתוח של התנהגות shoaling, אשר בדרך כלל כרוכות מדידת העדפת shoalmate, ניתן לבצע סוגים שונים של טנקים, עם שיטות שונות כדי לקבוע עמותת זמן 21-23. בפרוטוקול זה תת-סדרה ספציפית של overaרפרטואר assay ll התנהגותי מוצג. באופן ספציפי, פרוטוקול זה מציג מתודולוגיה כדי לעקוב אחר אנשים דרך תעוזה, תוקפנות, מבחני shoaling באופן המאפשר השוואות בתוך אנשים כדי לקבוע אם המושוות עקביות בכל יחידים בתוך אוכלוסייה. ביצענו פרוטוקול זה עם דג הזברה ציקלידים להרשיע (Amatitlania nigrofasciata) במחקרים קודמים 14,18, וזה יעבוד עם כל דג מים מתוקים בגודל דומה. מבחני תעוזה נערכים בתוך טנק טרפז 1.5 ליטר כי יש קו אופקי המגדירה אזורים בגודל שווה בחלקים העליונים ותחתונים של הטנק. התנהגויות לכמת כוללות את מספר מעברים ידי הדג הבדיקה בין החלק העליון והתחתון של הטנק, הזמן המושקע בכל חלק, מספר חצים, ואת חביון להיכנס החלק העליון. את assay התוקפנות מתבצע בתוך טנק מלבן 19L הכוללמראה אינץ 3 אינץ &#39;x 5 נוטה בסביבות 22 מעלות ממוקמות בפינה השמאלית התחתונה של הטנק 19. התנהגויות לכמת לכלול את הסכום הכולל של זמן מושקע על ידי דג יעד אינטראקציה עם המראה 20, יחד עם אינדיקטורים אגרסיווי ספציפית - מספר העקיצות, מציג לרוחב, חצים בין דגי המבחן וההשתקפות שלו. לקבלת מידע אודות מחוונים הספציפיים אלה, עקיצות מוגדרות מזנקים מהיר לעבר המראה עם פה פעור, מציג לרוחב מוגדר הלוהט של לרוחב, חזה, אנאליים הגבה סנפירים לכיוון המראה, וחיצים הם כל תנועות בלתי יציבות, כי הם לא מכוון במראה. לבסוף, assay shoaling מכמת את ההתנהגות של דג בדיקה בתא במרכז טנק תאי-תלת. תאי דופן האקווריום הם או ריקים, או להכיל &quot;נחיל יעד&quot; של דגים, ובפעם דגי המבחן מבלים ליד כל תא בצד נמדד 21-23. ציון כללי אחד, המכונה וויט&#39;לGTH של Shoaling (SOS), מחושב לגבי כל דג בדיקה אישית, ספציפית לגירויים, והוא יכול לשמש במורד זרם מנתח 14. כל ההתנהגויות הם הבקיעו ידי צופה יחיד, או צופה מרובה באמצעות תוכנת כימות התנהגותי חינם המכונית 24 JWatcher. בדיקת הנוכחות של תסמונת התנהגותית היא מאמץ סטטיסטי בעיקר, ומומלץ לעקוב אחר ההנחיות כפי שהוצג על ידי Budeav 2010 25. באופן ספציפי, מומלץ לבצע ניתוח מרכיבים עיקרי (PCA) על נתונים מרוכזים normed שבו תשומות הן הווקטורים של ההתנהגויות של פרט מבחנים עם מדידות התנהגותי מרובות (כלומר, תעוזה ותוקפנות). את PCA, שבוצעה על מטריצת מתאם, מפחיתה את ממדיות של המדידות התנהגותיות, ובכך מחלצת את הידע החשוב ביותר המסביר ברוב של הווריאציה. הרכיבים שחולצו לאחר מכן ניתן לפרש basאד על מקדמי מתאם גבוה עבור התנהגותם האישית של עשרות ריבית רגרסיה ניתן לחלץ עבור כל אדם על בסיס של רכיבים ההסבר. עשרות רגרסיה אלה לאחר מכן ניתן בהשוואה למדידה SOS ומדידות שאינם התנהגותיות שונות אחרות כגון גודל או מין דגים. עבודה זו יושמה במחקר של תסמונות התנהגותיות דג זברה שבה תסמונת ההתנהגותיות מין שקיימת בין תעוזת shoaling 14 התגלתה. במצב זה, גברי דג זברה נועזים נוטים יותר לקשר עם מינים אגרסיביים יותר, גדולים יותר (ד albolineatus), אך קשר זה הולך לאיבוד אצל נקבות. עבודה זו יושמה גם במחקר של קרובי משפחה לנוער להרשיע ציקליד (Amatitlania nigrofasciata) 18 שבה תסמונת התנהגותית לא התגלה, פוטנציאל המציין פלסטיות התנהגותית של מינים. לכן, בפרוטוקול הבא מוצג wה- i מטרה של המגדירה את האופי שלושה מבחנים ספציפיים (תעוזה, תוקפנות, shoaling) במסגרת לימוד תסמונת התנהגותית ברמת פרט.

<table><tbody><tr><td>Zebrafish Rack System</td><td>Aquaneering Inc</td><td>Cat. # ZS550</td><td></td></tr><tr><td>Pet Valu Tropical Fish Food, 224.0 g</td><td>Pet Valu</td><td>Cat. # 31700</td><td></td></tr><tr><td>Premium Grade Brine Shrimp Eggs, 16 oz</td><td>Brine Shrimp Direct</td><td></td><td></td></tr><tr><td>1.5 L Trapezoidal Tank</td><td>Pentair Aquatic Ecosystems</td><td>Cat. # itsts-a</td><td></td></tr><tr><td>19 L rectangular tank</td><td>That Fish Place</td><td>211932</td><td></td></tr><tr><td>76 L rectangular tank</td><td>That Fish Place</td><td>212180</td><td></td></tr><tr><td>Hitachi KP-D20A CCD Camera</td><td>Prescott&#039;s, Inc.</td><td></td><td></td></tr><tr><td>Nikon AF Nikkor 35-105 mm f/305~4.5s MACRO lens</td><td>Nikon Corporation</td><td></td><td></td></tr><tr><td>ArtMinds Square Mirror, Value Pack 3&quot; x 3&quot;</td><td>Michaels</td><td>Cat. # 10334162</td><td></td></tr><tr><td>Jwatcher</td><td></td><td></td><td></td></tr><tr><td>SPSS Statistics Base</td><td>IBM</td><td></td><td></td></tr><tr><td>R</td><td>The R Foundation</td><td></td><td></td></tr></tbody></table>

boldness, aggression, and shoaling assays for zebrafish behavioral syndromes

תסמונת התנהגותית מתקיימת כאשר התנהגויות מסוימות אינטראקציה תחת בהקשרים שונים. דג הזברה הייתה נבדקת במחקרים אחרונים וזה חשוב לתקנן פרוטוקולים כדי להבטיח ניתוחים ופרשנויות נכונים. במחקרים הקודמים שלנו, אנו מודדים תעוזה ידי ניטור סדרה של התנהגויות (זמן קרוב לפני שטח, חביון מעברים, מספר מעברים, חצים) בתוך טנק טרפז 1.5 ליטר. כמו כן, אנו לכמת תוקפנות על ידי עקיצות התבוננות, מציג לרוחב, חצים, וזמן ליד מראה נוטה בתוך טנק 19 L מלבני. על ידי חלוקת טנק 76 L לשלישים, גם בדקנו העדפות shoaling. את assay shoaling הוא assay להתאמה אישית והוא יכול להיות מותאם עבור השערות ספציפיות. עם זאת, פרוטוקולים עבור assay זה גם חייבים להיות אחידים, אך גמיש מספיק עבור התאמה אישית. במחקרים קודמים, תא הסוף היו או ריק, כלול 5 או 10 דג זברה, או 5 danios פנינה (ד albolineatus). ב folגעיית כתב יד, אנו מציגים פרוטוקול מפורט ונתוני נציגים שמלווים יישומים מוצלחים של הפרוטוקול, אשר יאפשר לשכפול ניסויי תסמונת התנהגותית.

בהתאם לאופי של המחקר, ופרוטוקול מסוים המועסק, תוצאות ברורות כמה אפשריות בניסוי תסמונות התנהגותיות. הטבלאות הבאות ודמויות, בהם מצוין, מעובדים מנתונים במחקר הקודם שלנו שפורסם בכתב העת ההתנהגות תהליכים 14 וכתב העת דג הזברה 17. כשההצעה (כמתוא?...

Watch this Scientific Journal Video about Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes at JoVE.com

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

תעוזה, תוקפנות, ואת מבחני Shoaling עבור תסמונות התנהגותיות דג הזברה

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

boldness-aggression-shoaling-assays-for-zebrafish-behavioral

Research

JoVE Journal

Medicine

10.2K Views.  Saint Joseph’s University. This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

Video: Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities 

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the visual scene. However, the OKR requires that an animal detect moving stripes and it is possible that fish that fail to exhibit an OKR may not be completely blind. One zebrafish mutant, the no optokinetic response c (nrc) has no OKR under any light conditions tested and was reported to be completely blind. Previously, we have shown that OFF-ganglion cell activity can be recorded in these mutants. To determine whether mutant fish with no OKR such as the nrc mutant can detect simple light increments and decrements we developed the visual motor behavioral assay (VMR). In this assay, single zebrafish larvae are placed in each well of a 96-well plate allowing the simultaneous monitoring of larvae using an automated video-tracking system. The locomotor responses of each larva to 30 minutes light ON and 30 minutes light OFF were recorded and quantified. WT fish have a brief spike of motor activity upon lights ON, known as the startle response, followed by return to lower-than baseline activity, called a freeze. WT fish also sharply increase their locomotor activity immediately following lights OFF and only gradually (over several minutes) return to baseline locomotor activity. The nrc mutants respond similarly to light OFF as WT fish, but exhibit a slight reduction in their average activity as compared to WT fish. Motor activity in response to light ON in nrc mutants is delayed and sluggish. There is a slow rise time of the nrc mutant response to light ON as compared to WT light ON response. The results indicate that nrc fish are not completely blind. Because teleosts can detect light through non-retinal tissues, we confirmed that the immediate behavioral responses to light-intensity changes require intact eyes by using the chokh (chk) mutants, which completely lack eyes from the earliest stages of development. In our VMR assay, the chk mutants exhibit no startle response to either light ON or OFF, showing that the lateral eyes mediate this behavior. The VMR assay described here complements the well-established OKR assay, which does not test the ability of zebrafish larvae to respond to changes in light intensities. Additionally, the automation of the VMR assay lends itself to high-throughput screening for defects in light-intensity driven visual responses.

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities

We developed the Visual-Motor Response to quantitate the motor output of larval zebrafish in response to light increments and decrements. We also examined zebrafish vision mutants, including the no optokinetic response (nrc) mutants, which were thought to be completely blind when tested by another vision assay, the optokinetic reflex.

Assay ההתנהגות כדי למדוד את התגובה של דג הזברה לשינויים לעוצמות אור

The optokinetic reflex (OKR) is a basic visual reflex exhibited by most vertebrates and plays an important role in stabilizing the eye relative to the ...

Biology

Behavioral Approaches to Studying Innate Stress in Zebrafish

Responding appropriately to stressful stimuli is essential for survival of an organism. Extensive research has been done on a wide spectrum of stress-related diseases and psychiatric disorders, yet further studies into the genetic and neuronal regulation of stress are still required to develop better therapeutics. The zebrafish provides a powerful genetic model to investigate the neural underpinnings of stress, as there exists a large collection of mutant and transgenic lines. Moreover, pharmacology can easily be applied to zebrafish, as most drugs can be added directly to water. We describe here the use of the 'novel tank test' as a method to study innate stress responses in zebrafish, and demonstrate how potential anxiolytic drugs can be validated using the assay. The method can easily be coupled with zebrafish lines harboring genetic mutations, or those in which transgenic approaches for manipulating precise neural circuits are used. The assay can also be used in other fish models. Together, the described protocol should facilitate the adoption of this simple assay to other laboratories.

This manuscript describes a simple method to measure stress behaviorally in adult zebrafish. The approach takes advantage of the innate tendency that zebrafish prefer the bottom half of a tank when in a stressful state. We also describe methods for coupling the assay with pharmacology.

גישות התנהגותיות לחקר הלחץ הטבוע בדג זברה

Responding appropriately to stressful stimuli is essential for survival of an organism. Extensive research has been done on a wide spectrum of ...

Behavior

Automated High-throughput Behavioral Analyses in Zebrafish Larvae

We have created a novel high-throughput imaging system for the analysis of behavior in 7-day-old zebrafish larvae in multi-lane plates. This system measures spontaneous behaviors and the response to an aversive stimulus, which is shown to the larvae via a PowerPoint presentation. The recorded images are analyzed with an ImageJ macro, which automatically splits the color channels, subtracts the background, and applies a threshold to identify individual larvae placement in the lanes. We can then import the coordinates into an Excel sheet to quantify swim speed, preference for edge or side of the lane, resting behavior, thigmotaxis, distance between larvae, and avoidance behavior. Subtle changes in behavior are easily detected using our system, making it useful for behavioral analyses after exposure to environmental toxicants or pharmaceuticals.

Our laboratory developed a novel high-throughput automated imaging system that is useful for the detection of several different behaviors in 7-day-old zebrafish larvae. The system can be used for detecting subtle changes in behavior after the larvae have been exposed to environmental toxicants or pharmaceuticals.

ניתוח התנהגות תפוקה גבוהה אוטומטי בזחלי דג הזברה

We have created a novel high-throughput imaging system for the analysis of behavior in 7-day-old zebrafish larvae in multi-lane plates. This system ...

Neuroscience

The Three-Chamber Choice Behavioral Task using Zebrafish as a Model System

Neurodegenerative diseases are age-dependent, debilitating, and incurable. Recent reports have also correlated hyperglycemia with changes in memory and/or cognitive impairment. We have modified and developed a three-chamber choice cognitive task similar to that used with rodents for use with hyperglycemic zebrafish. The testing chamber consists of a centrally located starting chamber and two choice compartments on either side, with a shoal of conspecifics used as the reward. We provide data showing that once acquired, zebrafish remember the task at least 8 weeks later. Our data indicate that zebrafish respond robustly to this reward, and we have identified cognitive deficits in hyperglycemic fish after 4 weeks of treatment. This behavioral assay may also be applicable to other studies related to cognition and memory.

We present a behavioral chamber designed to assess cognitive performance. We provide data showing that once acquired, zebrafish remember the task 8 weeks later. We also show that hyperglycemic zebrafish have altered cognitive performance, indicating that this paradigm is applicable to studies assessing cognition and memory.

המשימה ההתנהגותית של בחירה תלת-תאית באמצעות דג זברה כמערכת מודל

Neurodegenerative diseases are age-dependent, debilitating, and incurable. Recent reports have also correlated hyperglycemia with changes in memory and/or ...

Modeling Fetal Alcohol Spectrum Disorders in Zebrafish to Characterize the Impact of an Adverse Embryonic Environment on Adult Social Behavior

Fetal alcohol spectrum disorders (FASD) describe all alcohol-induced birth defects. Birth defects such as growth deficiencies, craniofacial, behavioral, and cognitive abnormalities are associated with FASD. Social difficulties are common behavioral abnormalities associated with FASD and often result in serious health issues. Animal models are critical to understanding the mechanisms responsible for ethanol-induced social defects. Zebrafish are social vertebrates that produce externally fertilized transparent eggs; these characteristics provide researchers with a precise yet simple procedure for creating the FASD phenotype and an innate behavior that can be leveraged to model the social deficits associated with FASD. Thus, zebrafish are ideal for characterizing the social deficits of FASD. The goal of the current protocol is to provide the user with a simple behavioral assay that can be used to characterize the consequences of a negative environment early during development and the effects it can have on social behavior in adulthood. The protocol can be used to characterize the effect mutations or teratogens have on adult social behavior. The protocol outlined here demonstrates how to characterize the social behavior of individual fish during a 20-min social assay. Furthermore, the data obtained using the current protocol provides evidence that the protocol can be used to characterize the effects of embryonic ethanol-induced social defects in adult zebrafish.

The goal of the current protocol is to outline the steps necessary to establish and use a social preference assay for adult zebrafish and demonstrate that it can be used to characterize ethanol-induced social defects.

Fetal alcohol spectrum disorders (FASD) describe all alcohol-induced birth defects. Birth defects such as growth deficiencies, craniofacial, behavioral, ...

Behavioral And Physiological Analysis In A Zebrafish Model Of Epilepsy

Epilepsy represents one of the most common neurological disorders, affecting an estimated 50 million people worldwide. Recent advances in genetic research have uncovered a large spectrum of genes implicated in various forms of epilepsy, highlighting the heterogeneous nature of this disorder. Appropriate animal models are essential for investigating the pathological mechanisms triggered by genetic mutations implicated in epilepsy and for developing specialized, targeted therapies. In recent years, zebrafish has emerged as a valuable vertebrate organism for modeling epilepsies, with the use of both genetic manipulation and exposure to known epileptogenic drugs, such as pentylenetetrazole (PTZ), to identify novel anti-epileptic therapeutics. Deleterious mutations in the mTOR regulator DEPDC5 have been associated with various forms of focal epilepsies and knock-down of the zebrafish orthologue causes hyperactivity associated with spontaneous seizure-like episodes, as well as enhanced electrographic activity and characteristic turn wheel swimming. Here, we described the method involved in generating the DEPDC5 loss-of-function model and illustrate the protocol for assessing motor activity at 28 and 48 h post fertilization (hpf), as well as a method for recording field activity in the zebrafish optic tectum. An illustration of the effect of the epileptogenic drug PTZ on neuronal activity over time is also provided.

Here, we present a protocol for the development and the characterization of a zebrafish model of epilepsy resulting from the transient inhibition of the DEPDC5 gene.

ניתוח התנהגותי ופיזיולוגי במודל דגי זברה של אפילפסיה

Epilepsy represents one of the most common neurological disorders, affecting an estimated 50 million people worldwide. Recent advances in genetic research ...

Using an Automated 3D-tracking System to Record Individual and Shoals of Adult Zebrafish

Like many aquatic animals, zebrafish (Danio rerio) moves in a 3D space. It is thus preferable to use a 3D recording system to study its behavior. The presented automatic video tracking system accomplishes this by using a mirror system and a calibration procedure that corrects for the considerable error introduced by the transition of light from water to air. With this system it is possible to record both single and groups of adult zebrafish. Before use, the system has to be calibrated. The system consists of three modules: Recording, Path Reconstruction, and Data Processing. The step-by-step protocols for calibration and using the three modules are presented. Depending on the experimental setup, the system can be used for testing neophobia, white aversion, social cohesion, motor impairments, novel object exploration etc. It is especially promising as a first-step tool to study the effects of drugs or mutations on basic behavioral patterns. The system provides information about vertical and horizontal distribution of the zebrafish, about the xyz-components of kinematic parameters (such as locomotion, velocity, acceleration, and turning angle) and it provides the data necessary to calculate parameters for social cohesions when testing shoals.

The use of a 3D automatic video system that can track individual and groups of zebrafish is described. As application example we explore the effects of the NMDA-receptor antagonist MK-801 on shoals of zebrafish.

Like many aquatic animals, zebrafish (Danio rerio) moves in a 3D space. It is thus preferable to use a 3D recording system to study its behavior. The ...

Analysis of Skeletal Muscle Defects in Larval Zebrafish by Birefringence and Touch-evoke Escape Response Assays

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular dystrophies1-3, congenital myopathies4,5, and disruptions in sarcomeric assembly6,7, due to high genomic and structural conservation with mammals8. Muscular disorganization and locomotive impairment can be quickly assessed in the zebrafish over the first few days post-fertilization. Two assays to help characterize skeletal muscle defects in zebrafish are birefringence (structural) and touch-evoked escape response (behavioral).
Birefringence is a physical property in which light is rotated as it passes through ordered matter, such as the pseudo-crystalline array of muscle sarcomeres9. It is a simple, noninvasive approach to assess muscle integrity in translucent zebrafish larvae early in development. Wild-type zebrafish with highly organized skeletal muscle appear very bright amidst a dark background when visualized between two polarized light filters, whereas muscle mutants have birefringence patterns specific to the primary muscular disorder they model. Zebrafish modeling muscular dystrophies, diseases characterized by myofiber degeneration followed by repeated rounds of regeneration, exhibit degenerative dark patches in skeletal muscle under polarized light. Nondystrophic myopathies are not associated with necrosis or regenerative changes, but result in disorganized myofibers and skeletal muscle weakness. Myopathic zebrafish typically show an overall reduction in birefringence, reflecting the disorganization of sarcomeres.
The touch-evoked escape assay involves observing an embryo&#39;s swimming behavior in response to tactile stimulation10-12. In comparison to wild-type larvae, mutant larvae frequently display a weak escape contraction, followed by slow swimming or other type of impaired motion that fails to propel the larvae more than a short distance12. The advantage of these assays is that disease progression in the same fish type can be monitored in vivo for several days, and that large numbers of fish can be analyzed in a short time relative to higher vertebrates.

The zebrafish is now an established and powerful tool for modeling muscular dystrophies, congenital myopathies, and related neuromuscular diseases. Birefringence and touch-evoked escape behavior are two common noninvasive assays used to determine the degree of muscular disorganization and locomotive impairment of zebrafish embryos during early development.

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular ...

מקבץ בדיקות התנהגותיות להערכת השפעות נוירוטוקסיות בדגי זברה בוגרים

Neurotoxicity Assessment in Adult Danio rerio using a Battery of Behavioral Tests in a Single Tank

The presence of neuropathological effects proved to be, for many years, the main endpoint for assessing the neurotoxicity of a chemical substance. However, in the last 50 years, the effects of chemicals on the behavior of model species have been actively investigated. Progressively, behavioral endpoints were incorporated into neurotoxicological screening protocols, and these functional outcomes are now routinely used to identify and determine the potential neurotoxicity of chemicals. Behavioral assays in adult zebrafish provide a standardized and reliable means to study a wide range of behaviors, including anxiety, social interaction, learning, memory, and addiction. Behavioral assays in adult zebrafish typically involve placing the fish in an experimental arena and recording and analyzing their behavior using video tracking software. Fish can be exposed to various stimuli, and their behavior can be quantified using a variety of metrics. The novel tank test is one of the most accepted and widely used tests to study anxiety-like behavior in fish. The shoaling and social preference tests are useful in studying the social behavior of zebrafish. This assay is particularly interesting since the behavior of the entire shoal is studied. These assays have proven to be highly reproducible and sensitive to pharmacological and genetic manipulations, making them valuable tools for studying the neural circuits and molecular mechanisms underlying behavior. Additionally, these assays can be used in drug screening to identify compounds that may be potential modulators of behavior.
We will show in this work how to apply behavioral tools in fish neurotoxicology, analyzing the effect of methamphetamine, a recreational drug, and glyphosate, an environmental pollutant. The results demonstrate the significant contribution of behavioral assays in adult zebrafish to the understanding of the neurotoxicological effects of environmental pollutants and drugs, in addition to providing insights into the molecular mechanisms that may alter neuronal function.

מחבר זרקור: הערכה של שינויים סביבתיים והתנהגותיים הנגרמים על ידי תרופות בדגי זברה בוגרים 

Here, we present a comprehensive behavioral test battery, including the novel tank, Shoaling, and social preference tests, to effectively determine the potential neurotoxic effects of chemicals (e.g., methamphetamine and glyphosate) on adult zebrafish using a single tank. This method is relevant to neurotoxicity and environmental research.