Name: alternative method of removing otoliths from sturgeon
Uploaded: 2016-06-27T15:00:00.000+00:00
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特别感谢瑞吉莫霍克环保部，国家韦拉卡鱼类孵化场，美国鱼类和野生动物局研究单位，南伊利诺伊大学，美国鱼类和野生动物服务东北渔业中心，英镑鱼子酱，美国加州大学戴维斯分校，加里森坝国家鱼类孵化场，熊崖国家鱼类孵化场和美国鱼类和野生动物服务巴拿马市水产资源局的帮助为这个项目提供的鲟鱼。 本文是美国地质调查局大湖科学中心的贡献2013。

重要提示:由于美国鲟所有物种的保护状态，允许收购和鱼之前，实验室方法得到处理。所有标本均从孵化来源获得验尸由于自然原因和处置下列标准程序。 1.试样制备<ol><li>前的任何切口侧放鲟胎体和用卷尺从鼻子到尾部精确到1mm的前端测量总长度。（图1A） </li><li>在配衡衡称重鲟鱼胴体至最接近克评估任何外部标记或标记和记录胴体。 </li></ol> 2.耳石提取<ol><li>使与颅骨的基极和第一背盾之间的大的圆角刀的垂直切口穿过所述主体的头部从主体（图1B）中分离出来。去除头，涂后氡头部所以口朝上（图1C）。 </li><li>通过使用圆角刀沿头骨（图1D）的腹侧切开取出口的软组织和腮。使从鼻子到颅骨完全平分头（图1E）的底部的前端的电的骨锯的切口。用手分开的两半，以暴露脑腔（图2A）。 </li><li>取下并丢弃用钳子查看半规管脑腔（图2B）内脑质（红色方框左边）（红色框右侧）。 </li><li>脑腔内有镊子（图2C）拆下半规管软骨。小心刺穿半规管软骨与钳子的前端，并删除耳石（图2D）。与来自半规管软骨镊子取出耳石（软骨内硬骨结构）（图2E） </stroNG&gt;。 </li><li>重复步骤2.3 - 2.4从头骨的另一半去除耳石。 </li><li>浸在92％乙醇的耳石，持续5秒，以消除任何软骨。在一个开设25毫升闪烁瓶中的地方耳石干燥24小时。 </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>

Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the U.S. Government.

提取鲟尸体耳石的另一种方法的概述已详细说明。要注意，特别注意的是需要在颅骨的中线的横向切口放置以均匀平分颅骨，以确保清扫期间不会伤害耳石是很重要的。如果切口是不够深以完全平分颅骨这将是非常困难的，以暴露脑腔，其中耳石的位置是清晰可见。有经验的，整体的技术是相对简单的，并导致被发现完整耳石的高百分比。这种新方法提供了研究的另一种方法，以提取比鲟可以使?...

鲟鱼（ 鲟 ）的人口在世界各地已经超过一个世纪的下降，由于影响，包括栖息地的丧失，人口分散，和过度捕捞，使得许多人口由州和联邦法律的保护1,2。管理机构在整个美国和世界已经确定鲟鱼为修复和恢复1目标物种。多数鲟鱼种群的人口特征（增长，招聘，饮食）进行了研究，以更好地了解基础生物学和生活史特点1。由于多数鲟鱼种群的保护状况，年龄评估是小受威胁居民在没有牺牲或损害个人是必要的困难。 骨结构的年龄估计和生长分析的目的，提取已常用的技术3。使用翅片射线段，以确定斯特奇的年龄上已成为一个实际非致命的方法，但该技术的验证已被限制和年龄可以是非常困难的老年个体（&gt; 15岁）破译4。研究人员评估在验证墨西哥湾鲟鱼（ 尖吻鲟desoti）5，大西洋鲟（A. O操作。oxyrinchus）6，白鲟（A. transmontanus）7，和湖鲟鱼的年龄中华鲟使用多个骨性结构及其使用（A. fluvescens）8总结说，第一边缘胸鳍线提供最准确的年龄，但结束了侵入性操作是有害的个体较大。目前，存在使用耳石形状指标成功9的非常高的速率北美鲟鱼种类区分的发展。与在颅底背横切口取出耳石的工艺细节是一种常见多耳石ction方法发现整个文献和塞科尔等清晰细腻。 （1991）10。然而，这种方法是难以在具有非常厚的头骨与小且脆弱和将趋向于打破耳石鲟实现。文学的进一步审查表明鲟鱼耳石去除过程中没有其他详细的方法。本文的目的是详细地从腹侧进入大脑腔提取鲟耳石的另一种方法。

<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

提取的鱼有很厚的头骨可能是困难和非常耗时的耳石（耳骨）。使得在用一只手或电锯头骨顶部的横纵切口如果操作不当可能损坏耳石的普遍做法。鲟鱼（ 鲟 ）是一个特别的家族，有一个非常大且厚头骨。从鱼揭露耳石的腹侧面进入大脑腔一个新的实验室方法比在文献中发现的其他耳石提取方法更容易。在文献中综述的方法被设计为场，并且在快速处理大量的鱼的更有效。但是，这种新的技术被设计为更适合于实验室环境时的时间并不紧迫，并且从每个试样成功提取是至关重要的。查找和使用这种技术去除耳石的成功是非常高的，且不会影响标tructure以任何方式。这种替代的技术是适用于其它类似的鱼种用于提取耳石。

世界各地的鲟鱼的成人的大小差别很大，但耳石的位置是一致的。使用一把锋利的片刀被发现删除图1D容易证明口腔的软组织。电动锯骨机是横向切口，显露图1E脑腔的首选工具。然而，注意平分头骨的中线需要否则耳石可以在此过程中压碎。如果正确进行该过程然后大脑空腔将均匀地为在图2A所示的耳石无损伤被暴露。脑物质容易与?...

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.

从鲟鱼去除耳石替代方法

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 ...

alternative-method-of-removing-otoliths-from-sturgeon

Research

JoVE Journal

Environment

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

视频: Alternative Method of Removing Otoliths from Sturgeon

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.

对水生生命值评价悬浮泥沙影响的流通系统曝光

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

科研

环境科学

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.

基于尸检的野生鱼类健康评估

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.

对河流生态系统中巨脊椎动物和鱼类的大小谱进行建模

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.

皮质醇从鱼翅和爪骨基质提取

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) ...

Extracellular Multi-Unit Recording from the Olfactory Nerve of Teleosts

Recent studies have shown that ocean acidification affects olfactory-driven behavior in fish. This may be due in part to a reduction in olfactory sensitivity in high PCO2/low pH water. To assess the effects of ocean acidification, or olfactory sensitivity in marine fish in general, we propose that extracellular multi-unit recording from the olfactory nerve is the method of choice. Although invasive, it is sensitive, robust, reproducible and independent of external salinity (unlike the electro-olfactogram [EOG], for example). Furthermore, it records a primary sensory input into the CNS, prior to any central processing. We show that this method can show a reduction in olfactory sensitivity that is both temporary and odorant-dependent, using a range of amino acids to construct concentration-response curves and calculate the thresholds of detection.

Extracellular multi-unit recording from the olfactory nerve is a sensitive, robust and reproducible method for assessing olfactory sensitivity in marine fish. It records primary sensory input and is independent of external salinity.

特勒奥茨的嗅觉神经的细胞外多单元记录

Recent studies have shown that ocean acidification affects olfactory-driven behavior in fish. This may be due in part to a reduction in olfactory ...

Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout (Oncorhynchus Mykiss) and Brown Trout (Salmo Trutta Morpha fario)

Egg collection is one of the most crucial procedures during fish reproduction in salmonid hatcheries. Classic methods involve the use of hand massage on fish abdomens to expel the eggs. An alternative method uses the pressure of gas injected into the body cavity, which causes the subsequent release of the eggs. This method is believed to have less negative effects on both the welfare and egg quality of the broodstocks. Herein, we compare the results of air and hand stripping methods with respect to one-year survival and egg quantity and quality in two salmonid fish, rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta morpha fario). Our results indicate that air stripping yielded a better quality of eggs and higher one-year survival rate in rainbow trout. In addition, air stripping resulted in lower mortality rate than the group subjected to hand stripping (25% vs. 35%). The pH and hatching rate of the hand stripped group was lower than those of the air stripped group. In the case of brown trout, the quality of eggs obtained by both hand and air-stripping methods was similar; however, the one-year losses in fish were higher in air stripped group (15% compared to 0% in hand stripped fish). Although the advantages of air stripping method over hand stripping in terms of egg quality might not be observed in all salmonid species, the air-stripping procedure might be a promising option to be adopted in hatcheries as it ensures a high level of reproducibility and efficiency.

Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout Oncorhynchus Mykiss and Brown Trout Salmo Trutta Morpha fario

The principal aim of this study is to standardize and test the pneumatic method (air stripping) of collecting eggs in rainbow trout and brown trout. This method allows effective and simple collection of the eggs without the necessity of fish abdomen massage.

两鲑鱼鱼、虹鳟 (虹鳟鱼虹鳟鱼) 和褐鳟 (鲑鲑形态石斑鱼) 的空气剥离效果

Egg collection is one of the most crucial procedures during fish reproduction in salmonid hatcheries. Classic methods involve the use of hand massage on ...

发育生物学

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.

使用计算机辅助视频和数字图像分析量化鱼游泳行为应对水铜的急性接触

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

神经科学

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.

从美国牛蛙的小囊（毛细胞的生理准备<em&gt;牛蛙</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.

解剖和三刺鳃骨架的平面安装

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

生物学

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka (Oryzias latipes)

Sex steroids, produced by the gonads, play an essential role in brain and pituitary tissue plasticity and in the neuroendocrine control of reproduction in all vertebrates by providing feedback to the brain and pituitary. Teleost fishes possess a higher degree of tissue plasticity and variation in reproductive&#160;strategies compared to mammals and appear to be useful models to investigate the role of sex steroids and the mechanisms by which they act. The removal of the main source of sex steroid production using gonadectomy together with blood sampling to measure steroid levels has been well-established and fairly feasible in bigger fish and is a powerful technique to investigate the role and effects of sex steroids. However, these techniques raise challenges when implemented in small size teleost models. Here, we describe the step-by-step procedures of gonadectomy in both males and female Japanese medaka followed by blood sampling. These protocols are shown to be highly feasible in medaka indicated by a high survival rate, safety for the life span and phenotype of the fish, and reproducibility in terms of sex steroid clearance. The use of these procedures combined with the other advantages of using this small teleost model will greatly improve the understanding of feedback mechanisms in the neuroendocrine control of reproduction and tissue plasticity provided by sex steroids in vertebrates.

Gonadectomy and Blood Sampling Procedures in the Small Size Teleost Model Japanese Medaka Oryzias latipes

The article describes a quick protocol to gonadectomize and sample blood from the small teleost fish, using Japanese medaka (Oryzias latipes) as a model, to investigate the role of sex steroids in animal physiology.

小尺寸远程模型日本美达卡（奥里齐亚斯拉蒂普斯）的果纳切除术和血液采样程序

Sex steroids, produced by the gonads, play an essential role in brain and pituitary tissue plasticity and in the neuroendocrine control of reproduction in ...