وقدمت التمويل لهذا البحث من قبل برنامج جائزة في علم الأعصاب من ولاية فلوريدا كلية الطب بجامعة وبدعم من وزارة العلوم البيولوجية في الاتحاد السوفيتي السابق. تم توفير التمويل الإضافي من قبل غرانت في والمعونة من صندوق أبحاث الكحول المشروبات المصنعة.

<ol>
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يعلن الكتاب أنه ليس لديهم مصالح مالية المتنافسة.

تكاليف تعاطي الكحول والإدمان على الكحول بالنسبة للمجتمع هائلة، سواء من حيث التكاليف البشرية والاقتصادية 30،31 29. ذبابة الفاكهة كنموذج يوفر نظام سريع وتنوعا لدراسة الاستجابات السلوكية بسرعة من عدد كبير من الأفراد وعلى هذا النحو وقد استخدم على نطاق واسع على ...

ذبابة الفاكهة السوداء البطن إظهار الاستجابات السلوكية للكحول ثنائي الطور (1) التي هي مماثلة لاستجابات الإنسان لهذا الدواء 2،3. عند التعرض الأولي لتركيزات منخفضة من الكحول، والذباب المعرض زيادة النشاط الحركي، والاستعاضة عن نقص في التنسيق الحركي، وفقدان السيطرة الوضعي والمقوم ردود الفعل (فقدان المنعكس التقويمي: LoRR)، والتخدير (الانعدام التام للنشاط الحركي في استجابة إلى التحفيز الميكانيكي) عن التعرض للكحول يتقدم 4-9. الساعة الإيقاعية الذاتية هو المغير قوية من حساسية الكحول وسمية كما لوحظ في الفئران 10،11، 12 الفئران، والبشر 13. وقد أظهرت التطورات الحديثة في مجال البحوث ذبابة الفاكهة على مدار الساعة الإيقاعية ينظم حساسية الكحول الحاد ولكن ليس الكحول التسامح 1. النهج الجينية القوية المتاحة في ذبابة الفاكهة من خلال دراسات متحولة والتلاعب المعدلة وراثيا من المكانيةوالزمانية التعبير الجيني توفير نظام يسمح التقدم السريع في تحديد الآليات الخلوية والجزيئية الكامنة لسلوكيات معقدة. استخدام ذبابة الفاكهة كأداة التحقيق سمحت بإحراز تقدم ملموس في فهم بيولوجيا الأعصاب الكحول التي يمكن أن تترجم بسرعة إلى الثدييات 14-16. من أجل تسهيل فهم الآليات الجزيئية التي من خلالها الساعة اليومية ينظم حساسية الكحول وبشكل موحد لقياس الاستجابات السلوكية عبر نقاط زمنية الإيقاعية، بروتوكول إدارة الكحول مناسبة للاستخدام في ظروف الإضاءة الخافتة الحمراء هو مطلوب. لذبابة الفاكهة، يمكن أن تدار من خلال مكملات غذائية الكحول التعرض المزمن أو موثوق من خلال ادارتها الكحول في شكل بخار عن التعرض الحاد. هنا، نحن تصف بروتوكول الإدارة الكحول مناسبة لتقييم تعديل الساعة البيولوجية للتخفيف من بين المنعكس التقويمي (LoRR) 1 وكذلكالتخدير. ومجرور الذباب مع 12 ساعة: 12 ساعة دورات LD عند درجة حرارة ثابتة، ثم نقل إلى نظام الضوء الخاضعة للرقابة ل2-5 أيام اعتمادا على مسألة تجريبية. ويتعرض الذباب لبخار الإيثانول في جهاز يعرف باسم FlyBar. في هذا الجهاز، وفقاعات كميات محددة من الهواء من خلال الماء والكحول؛ الأبخرة ثم يتم خلط وجهت إلى السكن قارورة الذباب. كل 5 دقائق الذباب وسجل للالرقم الذي فشل لعرض المقوم ردود الفعل أو أصبحت مخدرا. وتحسب النسب المئوية لLoRR كل نقطة زمنية ومقارنة بين نقاط الوقت الإيقاعية أو بين سلالات الذباب. بساطة وموثوقية التسليم الكحول باستخدام الكحول تسليم FlyBar جنبا إلى جنب مع خيارات التحليل السلوكي يوفر فائدة كبيرة للتجارب التي أجريت في ظل ظروف الإيقاعية الظلام.

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			<td height="21" style="height:21px;width:285px;">Alcohol 190 proof</td>
			<td style="width:115px;">Various</td>
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			<td height="21" style="height:21px;width:285px;">Name of Material</td>
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			<td height="21" style="height:21px;width:285px;">Aerator</td>
			<td style="width:115px;">Local pet store</td>
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			<td style="width:296px;">We use Whisper 60</td>
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			<td height="21" style="height:21px;width:285px;">Silicone tubing 1/8&#8221;</td>
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			<td style="width:119px;">408060-0030</td>
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			<td height="21" style="height:21px;width:285px;">120&#176; Y Connector</td>
			<td style="width:115px;">VWR</td>
			<td style="width:119px;">82017-256</td>
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			<td height="21" style="height:21px;width:285px;">Quick disconnects</td>
			<td style="width:115px;">VWR</td>
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			<td style="width:296px;"></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:285px;">Plastic tube clamps</td>
			<td style="width:115px;">Bell-art products</td>
			<td style="width:119px;">132250000</td>
			<td style="width:296px;">Either this or next</td>
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			<td style="width:296px;">Either this or previous</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Miniature Air Regulator Mounting Bracket</td>
			<td style="width:115px;">McMaster-Carr</td>
			<td style="width:119px;">9891K66</td>
			<td style="width:296px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Gilmont size 12 flow meter</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Tool clips</td>
			<td style="width:115px;">McMaster-Carr</td>
			<td style="width:119px;">1722A43</td>
			<td style="width:296px;">To hold flow meters</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Vial</td>
			<td style="width:115px;">VWR</td>
			<td style="width:119px;">89092-722</td>
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			<td height="21" style="height:21px;width:285px;">Rubber stopper with two holes</td>
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			<td style="width:119px;">59585-186</td>
			<td style="width:296px;">Fits in vials</td>
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			<td height="21" style="height:21px;width:285px;">5 mm Pyrex Glass tubes</td>
			<td style="width:115px;">Trikinetics</td>
			<td style="width:119px;">PGT5x65</td>
			<td style="width:296px;">Fits best in previous stopper.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Teflon tape</td>
			<td style="width:115px;">Hardware store</td>
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			<td style="width:296px;">To achieve snug fit in stoppers if necessary</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:285px;">Rubber stopper with two holes</td>
			<td style="width:115px;">VWR</td>
			<td style="width:119px;">59582-122</td>
			<td style="width:296px;">Fits our bottles</td>
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			<td style="width:296px;">Cut to length and bend by heating</td>
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			<td height="21" style="height:21px;width:285px;">Very fine nylon netting</td>
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			<td style="width:119px;">Various</td>
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			<td height="21" style="height:21px;width:285px;">15 watt bulbs</td>
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			<td style="width:296px;">Overhead red light</td>
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			<td height="21" style="height:21px;width:285px;">Photographic red safe light filters</td>
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		</tr>
	</tbody>
</table>

the flybar: administering alcohol to flies

ذبابة الفاكهة (ذبابة الفاكهة السوداء البطن) هي نموذج أنشئت لكلا البحوث الكحول والبيولوجيا الإيقاعية. في الآونة الأخيرة، وأظهرت لنا أن الساعة الإيقاعية ينظم حساسية الكحول، ولكن ليس تشكيل التسامح. هنا، نحن تصف بروتوكول لدينا في التفاصيل. يدار الكحول لالذباب باستخدام FlyBar. في هذا الإعداد، يتم خلط بخار الكحول المشبعة بالهواء مرطب في مجموعة النسب، وتدار على الذباب في أربعة أنابيب في وقت واحد. وتربى الذباب في ظل ظروف موحدة من أجل تقليل التباين بين مكررات. وتستخدم الذباب القديمة لمدة ثلاثة أيام من المورثات أو العلاجات المختلفة للتجارب، ويفضل عن طريق مطابقة الذباب من وجهتي نظر مختلفتين الوقت (على سبيل المثال، CT 5 و CT 17) إجراء مقارنات مباشرة ممكن. أثناء التجربة، ويتعرض الذباب لمدة 1 ساعة إلى نسبة محددة سلفا من بخار الكحول وعدد من الذباب الذي يحمل فقدان المنعكس التقويمي (LoRR) أو الحوار الاقتصادي الاستراتيجيتحسب أوجه كل 5 دقائق. ويمكن تحليل البيانات باستخدام ثلاثة أساليب إحصائية مختلفة. الأول هو لتحديد الوقت الذي 50٪ من الذباب فقدوا المنعكس المقوم واستخدام تحليل التباين (ANOVA) لتحديد ما إذا كانت توجد فروق ذات دلالة إحصائية بين نقاط الوقت. والثاني هو لتحديد النسبة المئوية التي تظهر الذباب LoRR بعد عدد معين من الدقائق، يعقبه تحليل ANOVA. الأسلوب الأخير هو تحليل كامل سلسلة مرات باستخدام الإحصاءات متعدد المتغيرات. ويمكن أيضا أن تستخدم بروتوكول للتجارب غير الإيقاعية أو المقارنات بين الأنماط الجينية.

تعديل الساعة البيولوجية من الكحول الحساسية باستخدام LoRR 50٪ كعلامة. ويرد مثال ممثل تظهر في التشكيل الإيقاعية حساسية الكحول أثناء النهار في الشكل 2. تم قياس LoRR في ست نقاط وقت خلال اليوم الثاني 2 من DD-S في كانتون وتم تحدي...

Watch this Scientific Journal Video about The FlyBar: Administering Alcohol to Flies at JoVE.com

The FlyBar: Administering Alcohol to Flies

وقد برزت ذبابة الفاكهة كنظام نموذج كبير للتشريح الأسس الخلوية والجزيئية من الاستجابات السلوكية للكحول. هنا نقدم بروتوكول لجمع البيانات حساسية الكحول في سياق الإيقاعية التي يمكن تطبيقها بسهولة على التجارب الأخرى ومناسبة تماما للبحوث الجامعية.

وFlyBar: إدارة الكحول الذباب

Fruit flies (Drosophila melanogaster) are an established model for both alcohol research and circadian biology. Recently, we showed that the circadian ...

the-flybar-administering-alcohol-to-flies

Research

JoVE Journal

Neuroscience

10.6K Views.  Florida State University.  وقد برزت ذبابة الفاكهة كنظام نموذج كبير للتشريح الأسس الخلوية والجزيئية من الاستجابات السلوكية للكحول. هنا نقدم بروتوكول لجمع البيانات حساسية الكحول في سياق الإيقاعية التي يمكن تطبيقها بسهولة على التجارب الأخرى ومناسبة تماما للبحو?...

Video: The FlyBar: Administering Alcohol to Flies

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and relapse. Various types of behavioral activity are commonly utilized as measures of drug motivation in animals. However, a crucial component of drug abuse relapse in abstinent cocaine users is "drug craving", which is difficult to model in animals, as it often occurs in the absence of overt behaviors. Yet, it is possible that a class of ultrasonic vocalizations (USVs) in rats may be a useful marker for affective responses to drug administration, drug anticipation and even drug craving. Rats vocalize in ultrasonic frequencies that serve as a communicatory function and express subjective emotional states. Several studies have shown that different call frequency ranges are associated with negative and positive emotional states. For instance, high frequency calls ("50-kHz") are associated with positive affect, whereas low frequency calls ("22-kHz") represent a negative emotional state. This article describes a procedure to assess rat USVs associated with daily cocaine self-administration. For this procedure, we utilized standard single-lever operant chambers housed within sound-attenuating boxes for cocaine self-administration sessions and utilized ultrasonic microphones, multi-channel recording hardware and specialized software programs to detect and analyze USVs. USVs measurements reflect emotionality of rats before, during and after drug availability and can be correlated with commonly assessed drug self-administration behavioral data such lever responses, inter-response intervals and locomotor activity. Since USVs can be assessed during intervals prior to drug availability (e.g., anticipatory USVs) and during drug extinction trials, changes in affect associated with drug anticipation and drug abstinence can also be determined. In addition, determining USV changes over the course of short- and long-term drug exposure can provide a more detailed interpretation of drug exposure effects on affective functioning.

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

تقييم الالفاظ بالموجات فوق الصوتية خلال مخدرات الذاتي الإدارة في الفئران

Drug self-administration procedures are commonly used to study behavioral and neurochemical changes associated with human drug abuse, addiction and ...

A Simple Way to Measure Ethanol Sensitivity in Flies

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain is correlated with that strain s ethanol preference, and therefore sedation is a highly relevant measure to study the genetics of alcohol responses and drinking. We demonstrate a simple way to expose flies to ethanol and measure its intoxicating effects. The assay we describe can determine acute sensitivity, as well as ethanol tolerance induced by repeat exposure. It does not require a technically involved setup, and can therefore be applied in any laboratory with basic fly culture tools.

A simple assay to measure the sedating effects of ethanol on Drosophila flies, based on the loss of righting reflex, is described.

طريقة بسيطة لقياس حساسية الايثانول في الذباب

Low doses of ethanol cause flies to become hyperactive, while high doses are sedating. The sensitivity to ethanol-induced sedation of a given fly strain ...

Vibrodissociation of Neurons from Rodent Brain Slices to Study Synaptic Transmission and Image Presynaptic Terminals

Mechanical dissociation of neurons from the central nervous system has the advantage that presynaptic boutons remain attached to the isolated neuron of interest. This allows for examination of synaptic transmission under conditions where the extracellular and postsynaptic intracellular environments can be well controlled. A vibration-based technique without the use of proteases, known as vibrodissociation, is the most popular technique for mechanical isolation. A micropipette, with the tip fire-polished to the shape of a small ball, is placed into a brain slice made from a P1-P21 rodent. The micropipette is vibrated parallel to the slice surface and lowered through the slice thickness resulting in the liberation of isolated neurons. The isolated neurons are ready for study within a few minutes of vibrodissociation. This technique has advantages over the use of primary neuronal cultures, brain slices and enzymatically isolated neurons including: rapid production of viable, relatively mature neurons suitable for electrophysiological and imaging studies; superior control of the extracellular environment free from the influence of neighboring cells; suitability for well-controlled pharmacological experiments using rapid drug application and total cell superfusion; and improved space-clamp in whole-cell recordings relative to neurons in slice or cell culture preparations. This preparation can be used to examine synaptic physiology, pharmacology, modulation and plasticity. Real-time imaging of both pre- and postsynaptic elements in the living cells and boutons is also possible using vibrodissociated neurons. Characterization of the molecular constituents of pre- and postsynaptic elements can also be achieved with immunological and imaging-based approaches.

This report demonstrates a technique for mechanical isolation of individual viable neurons retaining attached presynaptic boutons. Vibrodissociated neurons have the advantages of rapid production, excellent pharmacological control and improved space-clamp without influence from neighboring cells. This method can be used for imaging of synaptic elements and patch-clamp recording.

Vibrodissociation من الخلايا العصبية من الدماغ القوارض شرائح لدراسة نقل صورة متشابك ومحطات قبل المشبكي

Mechanical dissociation of neurons from the central nervous system has the advantage that presynaptic boutons remain attached to the isolated neuron of ...

Experimental Methods for Testing the Effects of Neurotrophic Peptide, ADNF-9, Against Alcohol-induced Apoptosis during Pregnancy in C57BL/6 Mice

Experimental designs for investigating the effects of prenatal alcohol exposure during early embryonic stages in fetal brain growth are challenging. This is mostly due to the difficulty of microdissection of fetal brains and their sectioning for determination of apoptotic cells caused by prenatal exposure to alcohol. The experiments described here provide visualized techniques from mice breeding to the identification of cell death in fetal brain tissue. This study used C57BL/6 mice as the animal model for studying fetal alcohol exposure and the role of trophic peptide against alcohol-induced apoptosis. The breeding consists of a 2-hr matting window to determine the exact stage of embryonic age. An established fetal alcohol exposure model has been used in this study to determine the effects of prenatal alcohol exposure in fetal brains. This involves free access to alcohol or pair-fed liquid diets as the sole source of nutrients for the pregnant mice.
The techniques involving dissection of fetuses and microdissection of fetal brains are described carefully, since the latter can be challenging. Microdissection requires a stereomicroscope and ultra-fine forceps. Step-by-step procedures for dissecting the fetal brains are provided visually. The fetal brains are dissected from the base of the primordium olfactory bulb to the base of the metencephalon.
For investigating apoptosis, fetal brains are first embedded in gelatin using a peel-away mold to facilitate their sectioning with a vibratome apparatus. Fetal brains embedded and fixed in paraformaldehyde are easily sectioned, and the free floating sections can be mounted in superfrost plus slides for determination of apoptosis or cell death.
TUNEL (TdT-mediated dUTP Nick End Labeling; TdT: terminal deoxynucleotidyl transferase) assay has been used to identify cell death or apoptotic cells. It is noteworthy that apoptosis and cell-mediated cytotoxicity are characterized by DNA fragmentation. Thus, the visualized TUNEL-positive cells are indicative of cell death or apoptotic cells.
The experimental designs here provide information about the use of an established liquid diet for studying the effects of alcohol and the role of neurotrophic peptides during pregnancy in fetal brains. This involves breeding and feeding pregnant mice, microdissecting fetal brains, and determining apoptosis. Together, these visual and textual techniques might be a source for investigating prenatal exposure of harmful agents in fetal brains.

The experimental designs proposed here focus on studying the effects of alcohol exposure in apoptosis and the application of neurotrophic peptide during pregnancy in fetal brain. A detailed description from the breeding to the collection of fetal brains is described. Techniques for determination of apoptosis are also described in detail.

الطرق التجريبية لاختبار آثار الببتيد عصبية، ADNF-9، ضد موت الخلايا المبرمج الناجم عن الكحول أثناء فترة الحمل في C57BL / 6 الفئران

Experimental designs for investigating the effects of prenatal alcohol exposure during early embryonic stages in fetal brain growth are challenging. This ...

Electroporation of the Hindbrain to Trace Axonal Trajectories and Synaptic Targets in the Chick Embryo

Electroporation of the chick embryonic neural tube has many advantages such as being quick and efficient for the expression of foreign genes into neuronal cells. In this manuscript we provide a method that demonstrates uniquely how to electroporate DNA into the avian hindbrain at E2.75 in order to specifically label a subset of neuronal progenitors, and how to follow their axonal projections and synaptic targets at much advanced stages of development, up to E14.5. We have utilized novel genetic tools including specific enhancer elements, Cre/Lox - based plasmids and the PiggyBac-mediated DNA transposition system to drive GFP expression in a subtype of hindbrain cells (the dorsal most subgroup of interneurons, dA1). Axonal trajectories and targets of dA1 axons are followed at early and late embryonic stages at various brainstem regions. This strategy contributes advanced techniques for targeting cells of interest in the embryonic hindbrain and for tracing circuit formation at multiple stages of development.

How neuronal networks are established in the embryonic brain is a fundamental question in developmental neurobiology. Here we combined an electroporation technique with novel genetic tools, such as Cre/Lox&#x2013;plasmids and PiggyBac-mediated DNA transposition system in the avian hindbrain to label dorsal interneurons and track their axonal projections and synaptic targets at various developmental stages.

Electroporation من الدماغ المؤخر لتتبع مسارات محور عصبي والأهداف متشابك في الجنين الفرخ

Electroporation of  the chick embryonic neural tube has many advantages such as being quick and efficient  for the expression of foreign genes into ...

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety of brain disorders1,2. To better understand the mechanisms by which brain circuits react to an animal's experience requires the ability to monitor the experience-dependent molecular changes in a given set of neurons, over a prolonged period of time, in the live animal. While experience and associated neural activity is known to trigger gene expression changes in neurons1,2, most of the methods to detect such changes do not allow repeated observation of the same neurons over multiple days or do not have sufficient resolution to observe individual neurons3,4. Here, we describe a method that combines in vivo two-photon microscopy with a genetically encoded fluorescent reporter to track experience-dependent gene expression changes in individual cortical neurons over the course of day-to-day experience. 
One of the well-established experience-dependent genes is Activity-regulated cytoskeletal associated protein (Arc)5,6. The transcription of Arc is rapidly and highly induced by intensified neuronal activity3, and its protein product regulates the endocytosis of glutamate receptors and long-term synaptic plasticity7. The expression of Arc has been widely used as a molecular marker to map neuronal circuits involved in specific behaviors3. In most of those studies, Arc expression was detected by in situ hybridization or immunohistochemistry in fixed brain sections. Although those methods revealed that the expression of Arc was localized to a subset of excitatory neurons after behavioral experience, how the cellular patterns of Arc expression might change with multiple episodes of repeated or distinctive experiences over days was not investigated. 
In vivo two-photon microscopy offers a powerful way to examine experience-dependent cellular changes in the living brain8,9. To enable the examination of Arc expression in live neurons by two-photon microscopy, we previously generated a knock-in mouse line in which a GFP reporter is placed under the control of the endogenous Arc promoter10. This protocol describes the surgical preparations and imaging procedures for tracking experience-dependent Arc-GFP expression patterns in neuronal ensembles in the live animal. In this method, chronic cranial windows were first implanted in Arc-GFP mice over the cortical regions of interest. Those animals were then repeatedly imaged by two-photon microscopy after desired behavioral paradigms over the course of several days. This method may be generally applicable to animals carrying other fluorescent reporters of experience-dependent molecular changes4.

In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons

Experience-dependent molecular changes in neurons are essential for the brain's ability to adapt in response to behavioral challenges. An in vivo two-photon imaging method is described here that allows the tracking of such molecular changes in individual cortical neurons through genetically encoded reporters.

في الجسم الحي ثنائي الفوتون التصوير من الخبرة التي تعتمد على التغيرات الجزيئية في الخلايا العصبية القشرية

The brain's ability to change in response to experience is essential for healthy brain function, and abnormalities in this process contribute to a variety ...

A Method to Make a Craniotomy on the Ventral Skull of Neonate Rodents

The use of a craniotomy for in vivo experiments provides an opportunity to investigate the dynamics of diverse cellular processes in the mammalian brain in adulthood and during development. Although most in vivo approaches use a craniotomy to study brain regions located on the dorsal side, brainstem regions such as the pons, located on the ventral side remain relatively understudied. The main goal of this protocol is to facilitate access to ventral brainstem structures so that they can be studied in vivo using electrophysiological and imaging methods. This approach allows study of structural changes in long-range axons, patterns of electrical activity in single and ensembles of cells, and changes in blood brain barrier permeability in neonate animals. Although this protocol has been used mostly to study the auditory brainstem in neonate rats, it can easily be adapted for studies in other rodent species such as neonate mice, adult rodents and other brainstem regions.

A surgical method is described to expose the ventral skull in neonate rats. Using this approach it is possible to open a craniotomy to perform acute electrophysiology and two-photon microscopy experiments in the brainstem of anesthetized pups.

وهناك طريقة لجعل حج القحف على بطني من الجمجمة الوليد القوارض

The use of a craniotomy for in vivo experiments provides an opportunity to investigate the dynamics of diverse cellular processes in the mammalian brain ...

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in their etiology. The fruit fly, Drosophila melanogaster, is a powerful model for exploring molecular-genetic mechanisms underlying alcohol-related behaviors and therefore holds great promise for identifying and understanding the function of genes that influence AUD. The use of the Drosophila model for these types of studies depends on the availability of assays that reliably measure behavioral responses to ethanol. This report describes an assay suitable for assessing ethanol sensitivity and rapid tolerance in flies. Ethanol sensitivity measured in this assay is influenced by the volume and concentration of ethanol used, a variety of previously reported genetic manipulations, and also the length of time the flies are housed without food immediately prior to testing. In contrast, ethanol sensitivity measured in this assay is not affected by the vigor of fly handling, sex of the flies, and supplementation of growth medium with antibiotics or live yeast. Three different methods for quantitating ethanol sensitivity are described, all leading to essentially indistinguishable ethanol sensitivity results. The scalable nature of this assay, combined with its overall simplicity to set-up and relatively low expense, make it suitable for small and large scale genetic analysis of ethanol sensitivity and rapid tolerance in Drosophila.

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

و، الفحص السلوكية قابلة غير مكلفة لقياس الإيثانول التخدير الحساسية والتسامح السريع في<em&gt; ذبابة الفاكهة</em

Alcohol use disorder (AUD) is a serious health challenge. Despite a large hereditary component to AUD, few genes have been unambiguously implicated in ...

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the development of more effective treatments is the relative lack of understanding of the molecular underpinnings of the effects of ethanol on behavior. The nematode, Caenorhabditis elegans (C.&#160;elegans), provides a useful model in which to generate and test hypotheses about the molecular effects of ethanol. Here, we describe an assay that has been developed and used to examine the roles of particular genes and environmental factors in behavioral responses to ethanol, in which locomotion is the behavioral output. Ethanol dose-dependently causes an acute depression of crawling on an agar surface. The effects are dynamic; animals exposed to a high concentration demonstrate an initial strong depression of crawling, referred to here as initial sensitivity, and then partially recover locomotion speed despite the continued presence of the drug. This ethanol-induced behavioral plasticity is referred to here as the development of acute functional tolerance. This assay has been used to demonstrate that these two phenotypes are distinct and genetically separable. The straightforward locomotion assay described here is suitable for examining the effects of both genetic and environmental manipulations on these acute behavioral responses to ethanol in C.&#160;elegans.

An Assay for Measuring the Effects of Ethanol on the Locomotion Speed of Caenorhabditis elegans

C. elegans is a useful model for studying the effects of ethanol on behavior. We present a behavioral assay that quantifies the effects of ethanol on the locomotion speed of crawling worms; both initial sensitivity and the development of acute functional tolerance to ethanol can be measured with this assay.

على الفحص لقياس آثار الإيثانول على سرعة تحرك من<em&gt; انواع معينة ايليجانس</em

Alcohol use disorders are a significant public health concern, for which there are few effective treatment strategies. One difficulty that has delayed the ...

Video Imaging and Spatiotemporal Maps to Analyze Gastrointestinal Motility in Mice

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central nervous system. Changes in ENS function are a major cause of GI symptoms and disease and may contribute to GI symptoms reported in neuropsychiatric disorders including autism. It is well established that isolated colon segments generate spontaneous, rhythmic contractions known as Colonic Migrating Motor Complexes (CMMCs). A procedure to analyze the enteric neural regulation of CMMCs in ex vivo preparations of mouse colon is described. The colon is dissected from the animal and flushed to remove fecal content prior to being cannulated in an organ bath. Data is acquired via a video camera positioned above the organ bath and converted to high-resolution spatiotemporal maps via an in-house software package. Using this technique, baseline contractile patterns and pharmacological effects on ENS function in colon segments can be compared over 3-4 hr. In addition, propagation length and speed of CMMCs can be recorded as well as changes in gut diameter and contraction frequency. This technique is useful for characterizing gastrointestinal motility patterns in transgenic mouse models (and in other species including rat and guinea pig). In this way, pharmacologically induced changes in CMMCs are recorded in wild type mice and in the Neuroligin-3R451C mouse model of autism. Furthermore, this technique can be applied to other regions of the GI tract including the duodenum, jejunum and ileum and at different developmental ages in mice.

This article describes a video imaging technique and high-resolution spatiotemporal mapping to identify changes in the neural regulation of colonic motility in adult mice. Subtle effects on gastrointestinal (GI) function can be detected using this approach in isolated tissue preparations to advance our understanding of GI disease.

التصوير الفيديو وخرائط الزمانية المكانية لتحليل الجهاز الهضمي على الحركة في الفئران

The enteric nervous system (ENS) plays an important role in regulating gastrointestinal (GI) motility and can function independently of the central ...