شكر خاص لشعبة سانت ريجيس الموهوك البيئة، Welaka الوطنية التفريخ السمك، السمك الولايات المتحدة وحدة أبحاث إدارة الحياة البرية، جامعة جنوب إلينوي، الولايات المتحدة للأسماك ومركز خدمة مصائد الأسماك في شمال الحياة البرية، والجنيه الاسترليني الكافيار، جامعة كاليفورنيا في ديفيز، حامية السد أسمال التفريخ، الدببة النصب الوطني تفريخ السمك، والسمك الولايات المتحدة ومكتب الموارد مدينة مصايد الأسماك الحياة البرية بنما لمساعدتهم توريد سمك الحفش لهذا المشروع، وهذا المقال هو المساهمة عام 2013 في مركز العلوم البحيرات العظمى هيئة المسح الجيولوجي الامريكية.

ملاحظة هامة: نظرا للوضع المحمية بجميع أنواع سمك الحفش في الولايات المتحدة، يسمح للحصول على والتعامل معها تم الحصول عليها الأسماك قبل إجراء المختبر. تم الحصول على جميع العينات من مصادر التفريخ بعد الوفاة لأسباب طبيعية والتخلص من اتباع إجراءات موحدة. 1. إعداد العينات <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> قبل أي شقوق تكمن الذبيحة سمك الحفش على جانبها وباستخدام شريط القياس لقياس الطول الكلي من الخطم إلى طرف الذيل إلى أقرب 1 مم. (الشكل 1A) </li><li style=";text-align:right;direction:rtl"> وزن سمك الحفش الذبيحة على نطاق tared إلى أقرب 1 غرام. تقييم الذبيحة لأية علامات خارجية أو علامات وسجل. </li></ol> 2. Otolith استخراج <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> جعل شق عمودي بسكين شرائح كبيرة بين قاعدة الجمجمة وأول ترس الظهرية من خلال الجسم لفصل الرأس عن الجسم (الشكل 1B). بعد إزالة الرأس، توآر الرأس بحيث يكون الفم هو مواجهة (الشكل 1C). </li><li style=";text-align:right;direction:rtl"> إزالة الأنسجة الناعمة في الفم والخياشيم باستخدام سكين شرائح لقطع على طول الجانب البطني من الجمجمة (1D الشكل). إجراء شق مع منشار العظام الكهربائي من طرف خطم إلى قاعدة الجمجمة إلى شطر تماما الرأس (الشكل 1E). فصل شطري باليد للكشف عن تجويف الدماغ (الشكل 2A). </li><li style=";text-align:right;direction:rtl"> إزالة والمواد الدماغ (المربع الأحمر يسار) مع ملقط لمشاهدة القنوات الهلالية (المربع الأحمر يمين) داخل تجويف الدماغ (الشكل 2B) تجاهل. </li><li style=";text-align:right;direction:rtl"> إزالة الغضروف الهلالي القناة من تجويف الدماغ مع ملقط (الشكل 2C). بعناية ثقب دائري الغضروف القناة مع غيض من ملقط وإزالة otolith (الشكل 2D). إزالة otolith (هيكل عظمي صلب داخل الغضروف) مع ملقط من الغضروف القناة الهلالية (الشكل 2E) </stroنانوغرام&gt;. </li><li style=";text-align:right;direction:rtl"> كرر الخطوات من 2،3-2،4 لإزالة otolith من النصف الآخر من الجمجمة. </li><li style=";text-align:right;direction:rtl"> تراجع otoliths في 92٪ من الإيثانول لمدة 5 ثانية لإزالة أي الغضروف. مكان otoliths في مفتوح التلألؤ قارورة 25 مل لتجف لمدة 24 ساعة. </li></ol>

<ol>
	<li>Auer, N. A., LeBreton, G. T. O., Beamish, F. W. H., McKinley, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conservation.">Conservation.</a> Sturgeons and Paddlefish of North America. 27, 252-276 (2004).</li><li>Campana, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy,+precision+and+quality+control+in+age+determination,+including+a+review+of+the+use+and+abuse+of+age+validation+methods).">Accuracy, precision and quality control in age determination, including a review of the use and abuse of age validation methods).</a> Journal of Fish Biology. 59, 197-242 (2001).</li><li>Bruch, R. M., Campana, S. E., Davis-Foust, S. L., Hansen, M. J., Janssen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lake+Sturgeon+age+validation+using+bomb+radiocarbon+and+known-age+fish.">Lake Sturgeon age validation using bomb radiocarbon and known-age fish.</a> Transactions of the American Fisheries Society. 138, 361-372 (2009).</li><li>Baremore, I. E., Rosati, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+validated,+minimally+deleterious+method+for+ageing+sturgeon.">A validated, minimally deleterious method for ageing sturgeon.</a> Fishery Bulletin. 112, 274-282 (2014).</li><li>Stevensen, J. T., Secor, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age+determination+and+growth+of+Hudson+River+Atlantic+sturgeon,+Acipenser+oxyrhynchus.">Age determination and growth of Hudson River Atlantic sturgeon, Acipenser oxyrhynchus.</a> Fishery Bulletin. 97, 153-166 (1999).</li><li>Brennan, J. S., Cailliet, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+age-determination+techniques+for+white+sturgeon+in+California.">Comparative age-determination techniques for white sturgeon in California.</a> Transactions of the American Fishery Society. 118, 296-310 (1989).</li><li>Rossiter, A., Noakes, D. L. G., Beamish, F. W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+age+estimation+for+the+lake+sturgeon.">Validation of age estimation for the lake sturgeon.</a> Transactions of the American Fisheries Society. 124, 777-781 (1995).</li><li>Chalupnicki, M. A., Dittman, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=North+American+Sturgeon+Otolith+Morphology.">North American Sturgeon Otolith Morphology.</a> Copeia. , 260-266 (2016).</li><li>Secor, D. H., Dean, J. M., Laban, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Manual for otolith removal and preparation for microstructural examination. , (1991).</li></ol>

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وقد فصلت محة عامة عن طريقة بديلة لاستخراج otoliths من جثث سمك الحفش. من المهم أن نلاحظ أن هناك حاجة إلى عناية خاصة لوضع شق الوحشي على خط الوسط من الجمجمة من أجل شطر الجمجمة بالتساوي لضمان عدم وجود ضرر على otoliths أثناء تشريح. إذا شق ليست عميقة بما يكفي لشطر تماما الجمجمة سيكون...

وقد سمك الحفش (Acipenseridae) السكان في جميع أنحاء العالم تتراجع لأكثر من قرن بسبب الآثار بما في ذلك فقدان الموائل، والتجزؤ السكان، والصيد الجائر مثل أن العديد من السكان ومحمية بموجب قوانين الدولة والحكومة الاتحادية 1،2. وقد حددت إدارة وكالات في جميع أنحاء الولايات المتحدة الأمريكية والعالم سمك الحفش كنوع الهدف لاستعادة واسترداد 1. وقد تم دراسة الخصائص السكانية (النمو والتوظيف والنظام الغذائي) لمعظم السكان سمك الحفش إلى فهم أفضل الأساسية البيولوجيا والتاريخ الحياة الخصائص 1. ويرجع ذلك إلى الوضع المحمي معظم السكان سمك الحفش، وتقييم سن صعب في السكان المهددين صغير حيث هناك ما يبرر أي تضحية أو ضرر للأفراد. وقد تم استخراج الهياكل العظمية لغرض تقدير العمر وتحليل النمو تقنية 3 المشتركة. استخدام لأقسام الأشعة الزعنفية لتحديد سن ستورجأصبح على طريقة غير قاتل العملي، ولكن تم التحقق من هذه التقنية محدودة والعمر يمكن أن يكون من الصعب جدا فك في الأفراد الأكبر سنا (&gt; 15 سنة) 4. الباحثون بتقييم استخدام هياكل عظمية متعددة في سمك الحفش لاستخدامها في التحقق من سن الخليج سمك الحفش (أسيبنسر oxyrinchus desoti) 5، والمحيط الأطلسي سمك الحفش (أ. س. oxyrinchus) 6، الأبيض سمك الحفش (A. ترانسمونتانوس) 7، وبحيرة سمك الحفش (أ fluvescens) 8 خلصت إلى أن أول الصدرية هامشية أشعة الزعانف قدم العمرية الأكثر دقة ولكن انتهى هذا الإجراء الغازية لتكون ضارة للأفراد أكبر. حاليا، هناك تطور استخدام مقاييس الشكل otolith للتمييز بين أنواع سمك الحفش في أمريكا الشمالية مع نسبة عالية جدا من النجاح 9. تفاصيل عن عملية إزالة otolith مع شق عرضي الظهري في قاعدة الجمجمة هي otolith المشترك خارجوجدت طريقة ction في جميع أنحاء الأدب ومفصلة بشكل واضح سيكور وآخرون. (1991) 10. ومع ذلك، وهذه الطريقة من الصعب تنفيذ على سمك الحفش التي لديها الجماجم سميكة جدا مع otoliths التي هي صغيرة وحساسة ومن شأنه أن يكسر. ويبين استعراض آخر للأدب لا منهجية تفصيلية أخرى لعملية إزالة otolith لسمك الحفش. وكان الهدف من هذه الورقة بالتفاصيل طريقة بديلة لاستخراج otoliths من سمك الحفش قبل دخول تجويف الدماغ من الجانب البطني.

<table><tbody><tr><td>Measuring Board</td><td></td><td></td><td>Metric meter stick is preferred</td></tr><tr><td>Weighing scale</td><td></td><td></td><td></td></tr><tr><td>Filleting Knife</td><td></td><td></td><td></td></tr><tr><td>Scalpel</td><td></td><td></td><td></td></tr><tr><td>Forceps</td><td></td><td></td><td></td></tr><tr><td>Sample vials</td><td></td><td></td><td>25 ml scintillation vials work well</td></tr></tbody></table>

alternative method of removing otoliths from sturgeon

استخراج otoliths (عظام الأذن) من الأسماك التي لها الجماجم سميكة جدا يمكن أن يكون الوقت الصعب وطويلا جدا. الممارسة الشائعة المتمثلة في جعل شق عمودي عرضية على الجزء العلوي من الجمجمة مع جهة أو منشار كهربائي قد يؤدي إلى تلف otolith إذا لم يتم تنفيذ بشكل صحيح. سمك الحفش (Acipenseridae) هي عائلة واحدة على وجه الخصوص أن يكون جمجمة كبيرة جدا وسميكة. كان هناك طريقة مختبر جديد دخول تجويف الدماغ من الجانب البطني من الأسماك لفضح otoliths أسهل من أساليب الاستخراج otolith أخرى وجدت في الأدب. تم تصميم الطرق المستعرضة في الكتابات عن هذا المجال وأكثر كفاءة في معالجة كميات كبيرة من الأسماك بسرعة. ومع ذلك، تم تصميم هذه التقنية الجديدة لتكون أكثر ملاءمة لإعداد مختبر عند الوقت ليس إلحاحا واستخراج ناجحة من كل عينة أمر بالغ الأهمية. نجاح إيجاد وإزالة otoliths باستخدام هذه التقنية عالية جدا ولا تفسد الصورةtructure في أي شكل من الأشكال. هذه التقنية البديلة هي التي تنطبق على أنواع الأسماك الأخرى المماثلة لاستخراج otoliths.

حجم الكبار من سمك الحفش في جميع أنحاء العالم يختلف كثيرا لكن موقع otoliths هو ثابت. تم العثور على استخدام سكين حاد شرائح لإزالة الأنسجة الرخوة في الفم هو مبين بسهولة في 1D الشكل. منشارا العظام الكهربائي هو الأداة المفضلة للشق الجانبي لفضح تجويف ?...

Watch this Scientific Journal Video about Alternative Method of Removing Otoliths from Sturgeon at JoVE.com

Alternative Method of Removing Otoliths from Sturgeon

The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses.

طريقة بديلة لإزالة Otoliths من سمك الحفش

Extracting the otoliths (ear bones) from fish that have very thick skulls can be difficult and very time consuming. The common practice of making a ...

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8.4K Views.  Great Lakes Science Center. The goal of the protocol is to show an effective method to extract the otoliths from sturgeon carcasses. 

Video: Alternative Method of Removing Otoliths from Sturgeon

A Time Differential Staining Technique Coupled with Full Bilateral Gill Denervation to Study Ionocytes in Fish

Branchial ionocytes (ICs) are the functional units for ionic regulation in fish. In adults, they are found on the filamental and lamellar epithelia of the gill where they transport ions such as Na+, Cl- and Ca2+ via a variety of ion channels, pumps and exchangers. The teleost gill is extrinsically innervated by the facial (VI), glossopharyngeal (IX) and vagus (X) nerves. The IX and X nerves are also the extrinsic source of branchial IC innervation. Here, two techniques used to study the innervation, proliferation and distribution of ICs are described: a time differential staining technique and a full bilateral gill denervation technique. Briefly, goldfish are exposed to a vital mitochondrion-specific dye (e.g., MitoTracker Red) which labels (red fluorescence) pre-existing ICs. Fish were either allowed to recover for 3 - 5 days or immediately underwent a full bilateral gill denervation. After 3 - 5 days of recovery, the gills are harvested and fixed for immunohistochemistry. The tissue is then stained with an &#945;-5 primary antibody (targets Na+/K+ ATPase containing cells) in conjunction with a secondary antibody that labels all (both new and pre-existing) ICs green. Using confocal imaging, it was demonstrated that pre-existing ICs appear yellow (labelled with both a viable mitochondrion-specific dye and &#945;-5) and new ICs appear green (labelled with &#945;-5 only). Both techniques used in tandem can be applied to study the innervation, proliferation and distribution of ICs on the gill filament when fish are exposed to environmental challenges.

This manuscript describes a protocol to track the re-distribution of branchial ionocytes and their innervation using a time differential staining technique coupled with full bilateral gill denervation.

تقنية وقت التفاضلية تلطيخ جانب كامل الثنائي جيل إزالة التعصيب لدراسة Ionocytes في السمك

Branchial ionocytes (ICs) are the functional units for ionic regulation in fish. In adults, they are found on the filamental and lamellar epithelia of the ...

Developmental Biology

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

نظام التعرض تدفق من خلال لتقييم موقوف الرواسب التأثيرات على الحياة المائية

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Necropsy-based Wild Fish Health Assessment

Anthropogenic influences from increased nutrients and chemical contaminants, to habitat alterations and climate change, can have significant effects on fish populations. Adverse effects monitoring, utilizing biomarkers from the organismal to the molecular level, can be used to assess the cumulative effects on fishes and other organisms. Fish health has been used worldwide as an indicator of aquatic ecosystem health. The necropsy-based fish health assessment provides data on visible abnormalities and lesions, parasites, condition and organosomatic indices. These can be compared by site, season and sex, as well as temporally, to document change over time. Severity ratings can be assigned to various observations to calculate a fish health index for more quantitative assessment. A drawback of the necropsy-based assessment is that it is based on visual observations and condition factors, which are not as sensitive as tissue and subcellular biomarkers for sublethal effects. Additionally, it is rarely possible to identify causes or risk factors associated with observed abnormalities. So, for instance a raised lesion or "tumor" on the fins, lips or body surface may be a neoplasm. However, it could also be a response to a parasite, chronic inflammation or hyperplasia of normal cells in response to an irritant. Conversely, neoplasms, certain parasites, other infectious agents and many tissue changes are not visible and so may be underestimated. However, during the necropsy-based assessment, blood (plasma), tissues for histopathology (microscopic pathology), genomics and other molecular analyses, and otoliths for aging can be collected. These downstream analyses, together with geospatial analyses, habitat assessments, water quality and contaminant analyses can all be important in comprehensive ecosystem evaluations.

The health of wild fishes can be used as an indicator of aquatic ecosystem health. Necropsy-based fish health assessments provide documentation of visible lesions or abnormalities, data used to calculate condition indices as well as the opportunity to collect tissues for microscopic evaluation, gene expression and other more in-depth analyses.

تقييم صحة الأسماك البرية المستندة إلى نيكروبسي

Anthropogenic influences from increased nutrients and chemical contaminants, to habitat alterations and climate change, can have significant effects on ...

Modeling the Size Spectrum for Macroinvertebrates and Fishes in Stream Ecosystems

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological community or food web. Importantly, the size spectrum assumes that individual size, rather than species&#8217; behavioral or life history characteristics, is the primary determinant of abundance within an ecosystem. Thus, unlike traditional allometric relationships that focus on species-level data (e.g., mean species&#8217; body size vs. population density), size spectra analyses are &#8216;ataxic&#8217; &#8211; individual specimens are identified only by their size, without consideration of taxonomic identity. Size spectra models are efficient representations of traditional, complex food webs and can be used in descriptive as well as predictive contexts (e.g., predicting responses of large consumers to changes in basal resources). Empirical studies from diverse aquatic ecosystems have also reported moderate to high levels of similarity in size spectra slopes, suggesting that common processes may regulate the abundances of small and large organisms in very different settings. This is a protocol to model the community-level size spectrum in wadable streams. The protocol consists of three main steps. First, collect quantitative benthic fish and invertebrate samples that can be used to estimate local densities. Second, standardize the fish and invertebrate data by converting all individuals to ataxic units (i.e., individuals identified by size, irrespective of taxonomic identity), and summing individuals within log2 size bins. Third, use linear regression to model the relationship between ataxic M and D estimates. Detailed instructions are provided herein to complete each of these steps, including custom software to facilitate D estimation and size spectra modeling.

This is a protocol to model the size spectrum (scaling relationship between individual mass and population density) for combined fish and invertebrate data from wadable streams and rivers. Methods include: field techniques to collect quantitative fish and invertebrate samples; lab methods to standardize the field data; and statistical data analysis.

نمذجة طيف الحجم للفقاريات والأسماك في النظم الإيكولوجية للتيار

The size spectrum is an inverse, allometric scaling relationship between average body mass (M) and the density (D) of individuals within an ecological ...

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) and quantify any differences in fin cortisol levels among three main sturgeon species. Fins were harvested from 19 sacrificed sturgeons including seven beluga (Huso huso), seven Siberian (Acipenser baerii), and five sevruga (A. stellatus). The sturgeons were raised in Iranian farms for 2 years (2017-2018), and cortisol extraction analysis was conducted in South Korea (January-February 2019). Jawbones from five H. huso were also used for cortisol extraction. Data were analyzed using the general linear model (GLM) procedure in the SAS environment. The intra- and inter-assay coefficients of variation were 14.15 and 7.70, respectively. Briefly, the cortisol extraction technique involved washing the samples (300 &#177; 10 mg) with 3 mL of solvent (ultrapure water and isopropanol) twice, rotation at 80&#160;rpm for 2.5 min, air-drying the washed samples at room temperature (22-28 &#176;C) for 7 days, further drying the samples using a bead beater at 50 Hz for 32 min and grinding them into powder, applying 1.5 mL methanol to the dried powder (75 &#177; 5 mg), and slow rotation (40 rpm) for 18 h at room temperature with continuous mixing. Following extraction, samples were centrifuged (9,500 x g for 10 min), and 1 mL supernatant was transferred into a new microcentrifuge tube (1.5 mL), incubated at 38 &#176;C to evaporate the methanol, and analyzed via enzyme-linked immunosorbent assay (ELISA). No differences were observed in fin cortisol levels among species or in fin and jawbone cortisol levels between washing solvents. The results of this study demonstrate that the sturgeon jawbone matrix is a promising alternative stress indicator to solid matrices.

In this study, we present a protocol for cortisol extraction from the fin and jawbone of sturgeon species. Fin and jawbone cortisol levels were further examined by comparing two washing solvents followed by ELISA assays. This study piloted the feasibility of jawbone cortisol as a novel stress indicator.

استخراج الكورتيزول من زعانف سمك الحفش ومصفوفات عظم الفك

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) ...

Streamlined Sampling and Cultivation of the Pelagic Cosmopolitan Larvacean, Oikopleura dioica

Oikopleura dioica is a planktonic chordate with exceptional filter-feeding ability, rapid generation time, conserved early development, and a compact genome. For these reasons, it is considered a useful model organism for marine ecological studies, evolutionary developmental biology, and genomics. As research often requires a steady supply of animal resources, it is useful to establish a reliable, low-maintenance culture system. Here we describe a step-by-step method for establishing an O. dioica culture. We describe how to select potential sampling sites, collection methods, target animal identification, and the set-up of the culturing system. We provide troubleshooting advice based on our own experiences. We also highlight critical factors that help sustain a robust culture system. Although the culture protocol provided here is optimized for O. dioica, we hope our sampling technique and culture setup will inspire new ideas for maintaining other fragile pelagic invertebrates.

Streamlined Sampling and Cultivation of the Pelagic Cosmopolitan Larvacean, Oikopleura dioica

Oikopleura dioica is a tunicate model organism in various fields of biology. We describe sampling methods, species identification, culturing setup, and culturing protocols for the animals and algal feed. We highlight key factors that helped strengthen the culture system and discuss the possible problems and resolutions.

تبسيط أخذ العينات وزراعة اليرقات العالمية الأوقيانوسية، أوكوبيورا دييكا

Oikopleura dioica is a planktonic chordate with exceptional filter-feeding ability, rapid generation time, conserved early development, and a compact ...

Patch Clamp Recordings in Inner Ear Hair Cells Isolated from Zebrafish

Patch clamp analyses of the voltage-gated channels in sensory hair cells isolated from a variety of species have been described previously1-4 but this video represents the first application of those techniques to hair cells from zebrafish. Here we demonstrate a method to isolate healthy, intact hair cells from all of the inner ear end-organs: saccule, lagena, utricle and semicircular canals. Further, we demonstrate the diversity in hair cell size and morphology and give an example of the kinds of patch clamp recordings that can be obtained. The advantage of the use of this zebrafish model system over others stems from the availability of zebrafish mutants that affect both hearing and balance. In combination with the use of transgenic lines and other techniques that utilize genetic analysis and manipulation, the cell isolation and electrophysiological methods introduced here should facilitate greater insight into the roles hair cells play in mediating these sensory modalities.

The purpose of this video is to demonstrate procedures for obtaining healthy, intact hair cells from the inner ear organs of adult zebrafish and then using them for patch clamp studies aimed at characterizing the biophysical properties of their voltage-gated channels.

تسجيلات المشبك التصحيح في الأذن الداخلية خلايا الشعر المعزولة من اسماك الزرد

Patch  clamp analyses of the voltage-gated channels in sensory hair cells isolated  from a variety of species have been described previously1-4 but this  ...

Neuroscience

Quantifying Fish Swimming Behavior in Response to Acute Exposure of Aqueous Copper Using Computer Assisted Video and Digital Image Analysis

Behavioral responses of aquatic organisms to environmental contaminants can be precursors of other effects such as survival, growth, or reproduction. However, these responses may be subtle, and measurement can be challenging. Using juvenile white sturgeon (Acipenser transmontanus) with copper exposures, this paper illustrates techniques used for quantifying behavioral responses using computer assisted video and digital image analysis. In previous studies severe impairments in swimming behavior were observed among early life stage white sturgeon during acute and chronic exposures to copper. Sturgeon behavior was rapidly impaired and to the extent that survival in the field would be jeopardized, as fish would be swept downstream, or readily captured by predators. The objectives of this investigation were to illustrate protocols to quantify swimming activity during a series of acute copper exposures to determine time to effect during early lifestage development, and to understand the significance of these responses relative to survival of these vulnerable early lifestage fish. With mortality being on a time continuum, determining when copper first affects swimming ability helps us to understand the implications for population level effects. The techniques used are readily adaptable to experimental designs with other organisms and stressors.

Measuring the impacts of environmental contaminants on fish behavior is often subjective and challenging particularly when dealing with sublethal endpoints. We describe methods including video technology to quantify swimming behavior of early life stage white sturgeon (Acipenser transmontanus) during and after 96 hr acute exposures to various concentrations of copper.

قياس سلوك الأسماك سباحة ردا على التعرض الحاد للمائي النحاس عن طريق الكمبيوتر بمساعدة الفيديو الرقمية وتحليل الصور

Behavioral responses of aquatic organisms to environmental contaminants can be precursors of other effects such as survival, growth, or reproduction.  ...

Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog (Rana catesbeiana)

The study of hearing and balance rests upon insights drawn from biophysical studies of model systems. One such model, the sacculus of the American bullfrog, has become a mainstay of auditory and vestibular research. Studies of this organ have revealed how sensory cells hair can actively detect signals from the environment. Because of these studies, we now better understand the mechanical gating and localization of a hair cell&#39;s transduction channels, calcium&#39;s role in mechanical adaptation, and the identity of hair cell currents. This highly accessible organ continues to provide insight into the workings of hair cells. Here we describe the preparation of the bullfrog&#39;s sacculus for biophysical studies on its hair cells. We include the complete dissection procedure and provide specific protocols for the preparation of the sacculus in specific contexts. We additionally include representative results using this preparation, including the calculation of a hair bundle&#39;s instantaneous force-displacement relation and measurement of a bundle&#39;s spontaneous oscillation.

Physiological Preparation of Hair Cells from the Sacculus of the American Bullfrog Rana catesbeiana

The American bullfrog&#39;s (Rana catesbeiana) sacculus permits direct examination of hair-cell physiology. Here the dissection and preparation of the bullfrog&#39;s sacculus for biophysical studies is described. We show representative experiments from these hair cells, including the calculation of a bundle&#39;s force-displacement relation and measurement of its unforced motion.

إعداد الفسيولوجية للخلايا الشعر من كييس من ضفدع الثور (<em&gt; رنا catesbeiana</em&gt;)

The study of hearing and balance rests upon insights drawn from biophysical studies of model systems. One such model, the sacculus of the American ...

Dissection and Flat-mounting of the Threespine Stickleback Branchial Skeleton

The posterior pharyngeal segments of the vertebrate head give rise to the branchial skeleton, the primary site of food processing in fish. The morphology of the fish branchial skeleton is matched to a species' diet. Threespine stickleback fish (Gasterosteus aculeatus) have emerged as a model system to study the genetic and developmental basis of evolved differences in a variety of traits. Marine populations of sticklebacks have repeatedly colonized countless new freshwater lakes and creeks. Adaptation to the new diet in these freshwater environments likely underlies a series of craniofacial changes that have evolved repeatedly in independently derived freshwater populations. These include three major patterning changes to the branchial skeleton: reductions in the number and length of gill raker bones, increases in pharyngeal tooth number, and increased branchial bone lengths. Here we describe a detailed protocol to dissect and flat-mount the internal branchial skeleton in threespine stickleback fish. Dissection of the entire three-dimensional branchial skeleton and mounting it flat into a largely two-dimensional prep allows for the easy visualization and quantification of branchial skeleton morphology. This dissection method is inexpensive, fast, relatively easy, and applicable to a wide variety of fish species. In sticklebacks, this efficient method allows the quantification of skeletal morphology in genetic crosses to map genomic regions controlling craniofacial patterning.

The branchial skeleton, including gill rakers, pharyngeal teeth, and branchial bones, serves as the primary site of food processing in most fish. Here we describe a protocol to dissect and flat-mount this internal skeleton in threespine sticklebacks. This method is also applicable to a variety of other fish species.

تشريح وشقة-تصاعد للThreespine أبو شوكة الخيشومي الهيكل العظمي

The posterior pharyngeal segments of the vertebrate head give rise to the branchial skeleton, the primary site of food processing in fish. The morphology ...