وأيد هذا العمل من قبل منحة من NHLBI لRW.

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H., Smith, H., Paddags, A., Hudson, R., Janssen, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+in+obesity+and+related+comorbid+conditions+after+diet-induced+weight+loss+or+exercise-induced+weight+loss+in+med+â&#x20AC;&#x201C;+A+randomized,+controlled+trial.">Reduction in obesity and related comorbid conditions after diet-induced weight loss or exercise-induced weight loss in med â&#x20AC;&#x201C; A randomized, controlled trial.</a> Annals of Internal Medicine. 133, 92-103 (2000).</li><li>Goodyea, L. J., Kahn, B. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise+Glucose+Transport,+and+Insulin+Sensitivity.">Exercise Glucose Transport, and Insulin Sensitivity.</a> Annual Review of Medicine. 49, 235-261 (1998).</li><li>Linke, A., Adams, V., Schulze, P. D., Erbs, S., Gielen, S., Fiehn, E., Mobius-Winkler, S., Schubert, A., Schuler, G., Hambrecht, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antioxidative+Effects+of+Exercise+Training+in+Patients+With+Chronic+Heart+Failure.">Antioxidative Effects of Exercise Training in Patients With Chronic Heart Failure.</a> Circulation. 111, 1763-1763 (2005).</li><li>Kemi, O. J., Ellingsen, O., Smith, G. L., Wisloff, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise-induced+changes+in+calcium+handling+in+left+ventricular+cardiomyocytes.">Exercise-induced changes in calcium handling in left ventricular cardiomyocytes.</a> Frontiers in Bioscience. 13, 356-368 (2008).</li><li>Wang, H., Hiatt, W. R., Barstow, T. J., Brass, E. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationships+between+muscle+mitochondrial+enzyme+activity+and+oxidative+capacity+in+man:+alterations+with+disease.">Relationships between muscle mitochondrial enzyme activity and oxidative capacity in man: alterations with disease.</a> Eur. J. Appl. Physiol. 80, 22-27 (1999).</li><li>Wu, Z., Puigserver, P., Andersson, U., Zhang, C., Adelmant, G., Mootha, V., Troy, A., Cinti, S., Lowell, B., Scarpulla, R. C., Spiegelman, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+Controlling+Mitochondrial+Biogenesis+and+Respiration+through+the+Thermogenic+Coactivator+PGC-1.">Mechanisms Controlling Mitochondrial Biogenesis and Respiration through the Thermogenic Coactivator PGC-1.</a> Cell. 98, 115-124 (1999).</li><li>Goto, M., Terada, S., Kato, M., Katoh, M., Yokozeki, T., Tabata, I., Shimokawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=cDNA+Cloning+and+mRNA+Analysis+of+PGC-1+in+Epirtrochlearis+Muscle+in+Swimming-Exercised+Rats.">cDNA Cloning and mRNA Analysis of PGC-1 in Epirtrochlearis Muscle in Swimming-Exercised Rats.</a> Biochemical and Biophysical Research Communications. , 274-350 (2000).</li><li>Piazza, N., Gosangi, B., Devilla, S., Arking, R., Wessells, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise-Training+in+Young+Drosophila+melanogaster+Reduces+Age-Related+Decline+in+Mobility+and+Cardiac+Performance.">Exercise-Training in Young Drosophila melanogaster Reduces Age-Related Decline in Mobility and Cardiac Performance.</a> PLoS ONE. 4, e5886-e5886 (2009).</li><li>Gargano, J. W., Martin, I., Bhandari, P., Grotewiel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+iterative+negative+geotaxis+(RING):+a+new+method+for+assessing+age-related+locomotor+decline+in+Drosophila.">Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila.</a> Exp Gerontol. 40, 386-395 (2005).</li></ol>

ليس لدينا ما يكشف.

وكان بروتوكول عام المعروضة هنا ناجحة في توثيق الآثار الفسيولوجية بعد التدريب. ومع ذلك، العديد من المناطق في هذا البروتوكول قد تخضع للتعديل لتناسب احتياجات تجريبية خاصة. على سبيل المثال، يمكن أن يحتمل أن تكون مدة التدريب وعدد من نوبات تكون متنوعة لجعل البرنامج أكثر أو أقل صعوبة، كما هو مطلوب. يمكن تعديل ارتفاع الحاوية التي يقاس بها قدرة انجذاب بالجاذبية السلبية لزيادة المساحة المتاحة للتحسين لتكون موثقة. قد الأساليب المختلفة لأتمتة تحديد الكميات من التسلق السرعة تكون أيضا قابلة للتطبيق. من حيث المبدأ، يمكن استخدام أي برنامج قادر على التمييز بين ذبابة من خلفية لتسريع عملية جمع البيانات. يجب أن يتم تعديل بعض جوانب البروتوكول إلا بحذر شديد، ولكن. على سبيل المثال، والتجارب الأولية تشير بقوة إلى أن ما لا يقل عن يوم واحد من الراحة في الأسبوع تميل إلى تيسير زيادة الظهورovement من ممارسة يومية لا هوادة فيها. بالإضافة إلى ذلك، ومن المعروف ايقاعات كل يوم، ودرجة الحرارة تؤثر على حركة الحيوانات ذات الدم البارد. يمكن أن تختلف الوقت من اليوم الذي يتم التدريب، ولكن ينبغي أن تكون متسقة دائما ضمن مجموعات خاصة في إطار المقارنة، من أجل تجنب احتمال آثار الخلط من ايقاعات كل يوم. التحكم في درجة الحرارة ومن الضروري أيضا، ونحن نوصي غرفة مخصصة عند درجة حرارة ثابتة لممارسة المعدات منزل. وأخيرا، لا بد من تربية الذكور والإناث، وتقاس على حدة، من أجل تجنب احتمال الخلط من آثار الخلافات خصوبة والجنس في القدرة على ممارسة الرياضة. وتقتصر التطبيقات المحتملة لهذه المنهجية إلا من خلال مخيلة الباحث. في الأعمال التمهيدية، واستخدمت هذه المنهجية أننا في ثلاثة تطبيقات واسعة: <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لقياس مدى تأثير تمارين التحمل في مختلف جوانب علم الأحياء النوع البري عبر العصور. </li><li style=";text-align:right;direction:rtl"> تيس قياس تأثير تمارين التحمل في الظواهر متحولة محددة. </li><li style=";text-align:right;direction:rtl"> للكشف عن عوامل وراثية والتي هي ضرورية لتنفيذ فوائد التمارين الرياضية </li></ol> كل من هذه التطبيقات تشمل مجموعة واسعة من إمكانيات محددة. بناء على خبرتنا الأولية، الظواهر متحولة تميل إلى التفاوت حسب مستوى ممارسة بقدر ما تختلف مع النظام الغذائي. استخدام نماذج اللافقارية إلى فهم أفضل للعلاقة بين ممارسة الرياضة، اتباع نظام غذائي، وعلم وظائف الأعضاء الشيخوخة وربما كان التطبيق الأكثر أهمية عام من هذا البروتوكول.

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> فهرس العدد </td><td> تعليقات </td></tr><tr><td> دايتون Gearmotor </td><td> غرينجر </td><td> 1LRA6A </td><td></td></tr><tr><td> RACO صندوق كهربائي </td><td> غرينجر </td><td> 5A052 </td><td></td></tr><tr><td> RACO الغطاء </td><td> غرينجر </td><td> 5A053 </td><td></td></tr><tr><td> كوبر بسمان </td><td> غرينجر </td><td> 6F043 </td><td></td></tr><tr><td> كوبر حامل بسمان </td><td> غرينجر </td><td> 1DD33 </td><td></td></tr><tr><td> كارلينج تقنيات تحويل </td><td> غرينجر </td><td> 2X464 </td><td></td></tr><tr><td> دايتون التحكم، AC / DC السرعة </td><td> غرينجر </td><td> 4X796 </td><td&gt; </td></tr><tr><td> Flugs للقوارير البلاستيكية تضييق نطاق </td><td> جينيسي العلمية </td><td> 49-102 </td><td></td></tr></tbody></table>

endurance training protocol and longitudinal performance assays for drosophila melanogaster

واحدة من المشاكل الأكثر إلحاحا التي تواجه الباحثين في مجال الطب الحديث هو المستويات المرتفعة من السمنة، مع زيادة لاحقة في الاضطرابات المرتبطة بها مثل السكري وأمراض القلب والأوعية الدموية 1-3. موضوع مهم للبحث في هذه المشاكل الصحية المرتبطة بها ينطوي على دور تمارين التحمل كتدخل مفيد. التمرينات هي غير مكلفة، وغير الغازية تدخل مع نتائج مفيدة عدة، بما في ذلك الحد في الجسم الدهون الزائدة 4، وزيادة الحساسية للانسولين في العضلات والهيكل العظمي 5، زادت الاستجابات المضادة للالتهابات ومضادات الأكسدة 6، وقدرة مقلص في تحسين العضلية 7. ومن المعروف انخفاض كثافة التمرين لزيادة نشاط الميتوكوندريا ونشوء حيوي في البشر (8) والفئران، مع coactivator النسخي PGC1-α كوسيط مهم 9،10. ما يتعلق بالفرعالشركة المصرية للاتصالات على أهمية ممارسة الرياضة كأداة لمكافحة العديد من المهم سن الأمراض ذات الصلة، وقد أعاق واسعة النطاق الدراسات الجينية طولية من عدم وجود بروتوكول التدريب على التحمل بالنسبة للأنواع التي لم تدم طويلا نموذج الجيني. مجموعة متنوعة من الأدوات الجينية المتاحة للاستخدام مع ذبابة الفاكهة، جنبا إلى جنب مع صيانة لها عمر قصير وغير مكلفة، وجعله نموذجا جذابا لمزيد من الدراسة لهذه الآليات الوراثية. مع هذا في الاعتبار وضعنا جهاز رواية، والمعروفة باسم برج الطاقة، على نطاق واسع لممارسة التدريب في الدروسوفيلا ذبابة الفاكهة 11. برج الطاقة يستخدم الذباب &quot;سلوك فطري انجذاب بالجاذبية سلبية متكررة للحث على التسلق السريع. في كل مرة مصاعد آلة، ثم يسقط، ومنهاج من الذباب، والتي يسببها الذباب في الصعود. الذباب مواصلة الاستجابة طالما أن الجهاز في العملية أو حتى تصبح مرهق للغاية للرد. وبالتالي، لا يمكن للباحث استخدام هذا الجهاز لتوفير قimultaneous التدريب لأعداد كبيرة من الذباب سن المطابقه ومطابقة وراثيا. بالإضافة إلى ذلك، ونحن تصف المقايسات المرتبطة مفيدة لتتبع التقدم المحرز طولية من الأفواج الطاير خلال التدريب.

Watch this Scientific Journal Video about Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster at JoVE.com

Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster

وصفنا أول بروتوكول التدريب على التحمل للحصول على نموذج الأنواع الجينية مهم،<em&gt; ذبابة الفاكهة السوداء البطن</em&gt;، والخطوط العريضة فحوصات عدة لرسم التحسينات في مجال التدريب التالية التنقل.

التحمل بروتوكول التدريب والمقايسات الأداء الطولية لذبابة الفاكهة الدروسوفيلا

One of the most pressing problems facing modern medical researchers is the surging levels of obesity, with the consequent increase in associated disorders ...

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Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster

15.5K Views.  University of Michigan Medical School. وصفنا أول بروتوكول التدريب على التحمل للحصول على نموذج الأنواع الجينية مهم، ذبابة الفاكهة السوداء البطن، والخطوط العريضة فحوصات عدة لرسم التحسينات في مجال التدريب التالية التنقل.

Video: Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

عالية الدقة تعقب الفيديو للتنقل في الكبار ذبابة الفاكهة السوداء البطن

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

Neurocircuit Assays for Seizures in Epilepsy Mutants of Drosophila

Drosophila melanogaster is a useful tool for studying seizure like activity. A variety of mutants in which seizures can be induced through either physical shock or electrical stimulation is available for study of various aspects of seizure activity and behavior. All flies, including wild-type, will undergo seizure-like activity if stimulated at a high enough voltage. Seizure like activity is an all-or-nothing response and each genotype has a specific seizure threshold. The seizure threshold of a specific genotype of fly can be altered either by treatment with a drug or by genetic suppression or enhancement. The threshold is easily measured by electrophysiology. Seizure-like activity can be induced via high frequency electrical stimulation delivered directly to the brain and recorded through the dorsal longitudinal muscles (DLMs) in the thorax. The DLMs are innervated by part of the giant fiber system. Starting with low voltage, high frequency stimulation, and subsequently raising the voltage in small increments, the seizure threshold for a single fly can be measured.

Neurocircuit Assays for Seizures in Epilepsy Mutants of Drosophila

Using high frequency electrical stimulation, seizure-like activity can be induced in Drosophila. This activity is easily recorded from the giant fiber system.

فحوصات Neurocircuit عن المضبوطات في المسوخ من الصرع ذبابة الفاكهة</em

Drosophila melanogaster is a useful tool for studying seizure like activity. A variety of mutants in which seizures can be induced through either physical ...

Microinjection Techniques for Studying Mitosis in the Drosophila melanogaster Syncytial Embryo

This protocol describes the use of the Drosophila melanogaster syncytial embryo for studying mitosis1. Drosophila has useful genetics with a sequenced genome, and it can be easily maintained and manipulated. Many mitotic mutants exist, and transgenic flies expressing functional fluorescently (e.g. GFP) - tagged mitotic proteins have been and are being generated. Targeted gene expression is possible using the GAL4/UAS system2. 

The Drosophila early embryo carries out multiple mitoses very rapidly (cell cycle duration, &asymp;10 min). It is well suited for imaging mitosis, because during cycles 10-13, nuclei divide rapidly and synchronously without intervening cytokinesis at the surface of the embryo in a single monolayer just underneath the cortex. These rapidly dividing nuclei probably use the same mitotic machinery as other cells, but they are optimized for speed; the checkpoint is generally believed to not be stringent, allowing the study of mitotic proteins whose absence would cause cell cycle arrest in cells with a strong checkpoint. Embryos expressing GFP labeled proteins or microinjected with fluorescently labeled proteins can be easily imaged to follow live dynamics (Fig. 1). In addition, embryos can be microinjected with function-blocking antibodies or inhibitors of specific proteins to study the effect of the loss or perturbation of their function3. These reagents can diffuse throughout the embryo, reaching many spindles to produce a gradient of concentration of inhibitor, which in turn results in a gradient of defects comparable to an allelic series of mutants. Ideally, if the target protein is fluorescently labeled, the gradient of inhibition can be directly visualized4. It is assumed that the strongest phenotype is comparable to the null phenotype, although it is hard to formally exclude the possibility that the antibodies may have dominant effects in rare instances, so rigorous controls and cautious interpretation must be applied. Further away from the injection site, protein function is only partially lost allowing other functions of the target protein to become evident.

Microinjection Techniques for Studying Mitosis in the Drosophila melanogaster Syncytial Embryo

This protocol describes the use of microinjection and high resolution imaging in the Drosophila melanogaster syncytial embryo to study mitosis.

تقنيات Microinjection لدراسة الانقسام الميتوزي في ذبابة الفاكهة السوداء البطن الأجنة المخلوي

This protocol describes the use of the Drosophila melanogaster syncytial embryo  for studying mitosis1.  Drosophila has useful genetics with a sequenced ...

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained and restrained whole larvae. For direct control of the environment around the heart another approach utilizes the dissected larvae and removal of the internal organs in order to bathe the heart in desired compounds. The exposed heart also allows membrane potentials to be monitored which can give insight of the ionic currents generated by the myocytes and for electrical conduction along the heart tube. These approaches have various advantages and disadvantages for future experiments that are discussed. The larval heart preparation provides an additional model besides the Drosophila skeletal NMJ to investigate the role of intracellular calcium regulation on cellular function. Learning more about the underlying ionic currents that shape the action potentials in myocytes in various species, one can hope to get a handle on the known ionic dysfunctions associated to specific genes responsible for various diseases in mammals.

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

رصد وظيفة القلب في اليرقات ذبابة الفاكهة السوداء البطن للدراسات فسيولوجية

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we present a protocol for the mounting of Drosophila melanogaster embryos and subsequent live imaging of fluorescently labeled hemocytes, the embryonic macrophages of this organism. Using the Gal4-uas system1 we drive the expression of a variety of genetically encoded, fluorescently tagged markers in hemocytes to follow their developmental dispersal throughout the embryo. Following collection of embryos at the desired stage of development, the outer chorion is removed and the embryos are then mounted in halocarbon oil between a hydrophobic, gas-permeable membrane and a glass coverslip for live imaging. In addition to gross migratory parameters such as speed and directionality, higher resolution imaging coupled with the use of fluorescent reporters of F-actin and microtubules can provide more detailed information concerning the dynamics of these cytoskeletal components.

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Drosophila hemocytes disperse over the entirety of the developing embryo. This protocol demonstrates how to mount and image these migrations using embryos with fluorescently labelled hemocytes.

يعيش تصوير ذبابة الفاكهة السوداء البطن هجرات كرية دموية الجنينية

Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we ...

Isolation of Drosophila melanogaster Testes

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline interactions. Testes are typically isolated from adult males 0-3 days after eclosion from the pupal case. The testes of wild-type flies are easily distinguished from other tissues because they are yellow, but the testes of white mutant flies, a common genetic background for laboratory experiments are similar in both shape and color to the fly gut. Performing dissection on a glass microscope slide with a black background makes identifying the testes considerably easier. Testes are removed from the flies using dissecting needles. Compared to protocols that use forceps for testes dissection, our method is far quicker, allowing a well-practiced individual to dissect testes from 200-300 wild-type flies per hour, yielding 400-600 testes. Testes from white flies or from mutants that reduce testes size are harder to dissect and typically yield 200-400 testes per hour.

Isolation of Drosophila melanogaster Testes

Drosophila melanogaster testes can be rapidly and efficiently isolated from adult males using dissecting needles. With practice, one can readily isolate in one or two days an amount of testes sufficient for the analysis of DNA or RNA by high throughput sequencing or more traditional molecular biology methods or of protein for antibody- or enzyme-based assays.

عزلة ذبابة الفاكهة السوداء البطن الخصيات

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline ...

Quantitative Comparison of cis-Regulatory Element (CRE) Activities in Transgenic Drosophila melanogaster

Gene expression patterns are specified by cis-regulatory element (CRE) sequences, which are also called enhancers or cis-regulatory modules. A typical CRE possesses an arrangement of binding sites for several transcription factor proteins that confer a regulatory logic specifying when, where, and at what level the regulated gene(s) is expressed. The full set of CREs within an animal genome encodes the organism&prime;s program for development1, and empirical as well as theoretical studies indicate that mutations in CREs played a prominent role in morphological evolution2-4. Moreover, human genome wide association studies indicate that genetic variation in CREs contribute substantially to phenotypic variation5,6. Thus, understanding regulatory logic and how mutations affect such logic is a central goal of genetics. 
Reporter transgenes provide a powerful method to study the in vivo function of CREs. Here a known or suspected CRE sequence is coupled to heterologous promoter and coding sequences for a reporter gene encoding an easily observable protein product. When a reporter transgene is inserted into a host organism, the CRE&prime;s activity becomes visible in the form of the encoded reporter protein. P-element mediated transgenesis in the fruit fly species Drosophila (D.) melanogaster7 has been used for decades to introduce reporter transgenes into this model organism, though the genomic placement of transgenes is random. Hence, reporter gene activity is strongly influenced by the local chromatin and gene environment, limiting CRE comparisons to being qualitative. In recent years, the phiC31 based integration system was adapted for use in D. melanogaster to insert transgenes into specific genome landing sites8-10. This capability has made the quantitative measurement of gene and, relevant here, CRE activity11-13 feasible. The production of transgenic fruit flies can be outsourced, including phiC31-based integration, eliminating the need to purchase expensive equipment and/or have proficiency at specialized transgene injection protocols. 
Here, we present a general protocol to quantitatively evaluate a CRE&prime;s activity, and show how this approach can be used to measure the effects of an introduced mutation on a CRE&prime;s activity and to compare the activities of orthologous CREs. Although the examples given are for a CRE active during fruit fly metamorphosis, the approach can be applied to other developmental stages, fruit fly species, or model organisms. Ultimately, a more widespread use of this approach to study CREs should advance an understanding of regulatory logic and how logic can vary and evolve.

Quantitative Comparison of cis-Regulatory Element CRE Activities in Transgenic Drosophila melanogaster

Phenotypic variation for traits can result from mutations in cis-regulatory element (CRE) sequences that control gene expression patterns. Methods derived for use in Drosophila melanogaster can quantitatively compare the levels of spatial and temporal patterns of gene expression mediated by modified or naturally occurring CRE variants.

الكمية مقارنة رابطة الدول المستقلة التنظيم العنصر (لجنة المساواة العرقية) في الأنشطة المعدلة وراثيا ذبابة الفاكهة السوداء البطن</em

Gene expression patterns are specified by cis-regulatory element (CRE) sequences, which are also called enhancers or cis-regulatory modules. A typical CRE ...

Measurement of Lifespan in Drosophila melanogaster

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced physical performance and increased risk of disease. Individual aging is manifest at the population level as an increase in age-dependent mortality, which is often measured in the laboratory by observing lifespan in large cohorts of age-matched individuals. Experiments that seek to quantify the extent to which genetic or environmental manipulations impact lifespan in simple model organisms have been remarkably successful for understanding the aspects of aging that are conserved across taxa and for inspiring new strategies for extending lifespan and preventing age-associated disease in mammals.
The vinegar fly, Drosophila melanogaster, is an attractive model organism for studying the mechanisms of aging due to its relatively short lifespan, convenient husbandry, and facile genetics. However, demographic measures of aging, including age-specific survival and mortality, are extraordinarily susceptible to even minor variations in experimental design and environment, and the maintenance of strict laboratory practices for the duration of aging experiments is required. These considerations, together with the need to practice careful control of genetic background, are essential for generating robust measurements. Indeed, there are many notable controversies surrounding inference from longevity experiments in yeast, worms, flies and mice that have been traced to environmental or genetic artifacts1-4. In this protocol, we describe a set of procedures that have been optimized over many years of measuring longevity in Drosophila using laboratory vials. We also describe the use of the dLife software, which was developed by our laboratory and is available for download (<a href="http://sitemaker.umich.edu/pletcherlab/software">http://sitemaker.umich.edu/pletcherlab/software</a>). dLife accelerates throughput and promotes good practices by incorporating optimal experimental design, simplifying fly handling and data collection, and standardizing data analysis. We will also discuss the many potential pitfalls in the design, collection, and interpretation of lifespan data, and we provide steps to avoid these dangers.

Measurement of Lifespan in Drosophila melanogaster

Drosophila melanogaster is a powerful model organism for exploring the molecular basis of longevity regulation. This protocol will discuss the steps involved in generating a reproducible, population-based measurement of longevity as well as potential pitfalls and how to avoid them.

قياس عمر في<em&gt; ذبابة الفاكهة الدروسوفيلا</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Maintenance of a Drosophila melanogaster Population Cage

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Maintenance of a Drosophila melanogaster Population Cage

This manuscript reports a detailed protocol for culturing, on a regular basis, a population of Drosophila melanogaster using a fly population cage.

صيانة<em&gt; ذبابة الفاكهة</em&gt; عدد السكان قفص

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life ...

Design and Development of Aptamer&#8211;Gold Nanoparticle Based Colorimetric Assays for In-the-field Applications

The design and development of an aptamer&#8211;gold nanoparticle (AuNP) colorimetric assay for the detection of small molecules for in-the-field applications was examined. Target selective AuNP based color assays have been developed in controlled proof-of-concept laboratory settings. However, these schemes have not been exerted to a point of failure to determine their practical use beyond laboratory settings. This work describes a generic approach to design, develop, and troubleshoot an aptamer-AuNP colorimetric assay for small molecule analytes and using the assay for in-the-field settings. The assay is advantageous because adsorbed aptamers passivate the nanoparticle surfaces and provide a means to reduce and eliminate false positive responses to non-target analytes. Transitioning this system to practical uses required defining not only the shelf-life of the aptamer-AuNP assay, but establishing methods and procedures for extending the long-term storage capabilities. Also, one of the recognized concerns with colorimetric readout is the burden placed on analysts to accurately identify often subtle changes in color. To lessen the responsibility on analysts in the field, a color analysis protocol was designed to perform the color identification duties without the need for performing this task on laboratory grade equipment. The method for creating and testing the data analysis protocol is described. However to understand and influence the design of adsorbed aptamer assays, the interactions associated with the aptamer, target, and AuNPs require further study. The knowledge gained could lead to tailoring aptamers for improved functionality.

The design and development of an aptamer&#8211;gold nanoparticle colorimetric assay for the detection of small molecules for in-the-field applications was examined. Additionally, a smart-device colorimetric application (app) was validated and long-term storage of the assay was established for use in the field.