We are grateful to anonymous reviewers for comments and suggestions that greatly improved this manuscript. We thank R. Bowes, R. Carreno, and T. Panhuis for assistance in collecting the fish. We also thank M. Lee for assistance with male preference trials. We are grateful to the Ohio Wesleyan University Department of Zoology for helpful advice and suggestions throughout this study and Arizona State University (McGraw Lab) for software advice.

Todos os experimentos foram realizados com a aprovação do Comitê de Cuidado e Uso Institucional Animal da Ohio Wesleyan University. 1. Gravação UV refletância dos peixes usando um espectrofotômetro <ol><li> Calibrar um espectrofotômetro e fonte de luz com um padrão branco conhecido em toda a gama de comprimentos de onda a ser medido de acordo com as recomendações do instrumento ou software. </li><li> Anestesiar os peixes, colocando em uma solução a 0,5% de acetato de 3 aminobenzoate-metano sal de ácido sulfônico (MS-222) tamponada com igual quantidade de bicarbonato de sódio até que o peixe deixa de responder. Uma vez que não responde, imediatamente colocar o peixe em um fundo preto, não-reflexivo. Nota: MS-222 é potencialmente tóxico, e devem ser usadas luvas em todos os momentos. </li><li> Fazer todas as medições em uma sala escura para minimizar o excesso de luz além do que a fonte de luz espectrômetro. </li><li> reflectância medida usando uma sonda de fibra óptica mantida contra o corpo do organismo. FaçoCertifique-se para formar um ângulo de 45 ° consistente entre as fibras ópticas e do corpo, como é construído em alguns modelos de sonda. Nota: Para manter a consistência de medição, pode ser desejável a média em vários scans (2 - 5) por região do corpo. </li><li> Gravar dados de reflectância com software de análise de reflectância apropriada. Nota: O software utilizado aqui foi definido como: integração em tempo = 2,000 seg, média scans = 2, e de pixel suavização = 5. Os dados foram copiados e colados em uma planilha. </li><li> Repetir em várias áreas do corpo para determinar a presença ou ausência de reflectância de UV e gama de variação fenotípica entre organismos. Nota: exames preliminares podem ser necessários para determinar qual, se houver, áreas do corpo mostram consistentemente marcações UV. </li><li> Devolver o peixe para um aquário de exploração bem oxigenado até que esteja totalmente recuperado, como evidenciado por batidas opérculo normais e comportamentos de natação. Nota: um protector comercial para restaurar o revestimento mucoso para o corpo podem ser adicionados para ajudar na recuperação da pele. </li><li> Repcomer em vários indivíduos, incluindo ambos os sexos, se desejar. </li><li> Na planilha, onde todos os dados foram copiados, os dados do gráfico usando um gráfico de dispersão, com reflectância no eixo Y e comprimento de onda no eixo-X. picos de amplitude na gama dos UV (300-400 nm) indicar a presença de reflectância de UV sobre o peixe. </li></ol> 2. Os ensaios de acasalamento dicotômicas Nota: Execute observações de controle, em que os peixes focal, filtros UV e filtros opacos estão no local. Nestes ensaios, quando os filtros são removidos opacas, peixes focal será exposto a partes do tanque indicam a presença e ausência de luz UV, mas que não possuem parceiros de acasalamento potenciais. Para mais detalhes sobre a validade dos testes de escolha dicotômicas neste experimento ver Palmer e Hankison (2015) 10, embora note que estes testes também têm desvantagens 11. <ol><li> Use indivíduos sexualmente receptivas como organismos focais. Use feminina P. latipinna &lt;48 horas pós-parto 5 aND machos isolado para, pelo menos, 24 horas antes dos testes para melhorar a probabilidade de motivação sexual. </li><li> Coloque um único indivíduo focal em uma área neutra adjacente a dois compartimentos de escolha. Para P. latipinna, usar um aquário teste 75,7 L (30,5 cm x 30,5 cm x 75 cm; Figura 1), dividido em três secções por meio de divisórias de Plexiglas transparente, formando uma área central para o indivíduo focal flanqueado por dois compartimentos de igual tamanho de escolha. Certifique-se de que nenhuma água ou olfativas pistas são compartilhadas entre as seções de tanques individuais. Repita o procedimento para cada indivíduo a ser observado. </li></ol><img alt="figura 1" src="/files/ftp_upload/54453/54453fig1.jpg" /> Figura 1. Experimental Aquarium Setup Um aquário retangular dividido em três seções:. Uma área central que realizou o indivíduo focal e dois compartimentos finais que realizou pares de objetos (a figura mostra uma mulher no meio com machos de objeto nas extremidades). fil intercambiáveisters que ou reflectância UV bloqueado ou permitido poderia ser colocada sobre o divisórias de vidro que dividem as seções masculina e feminina do aquário. Esta figura é modificada a partir Palmer e Hankison (2015) 10. <a href="http://ecsource.jove.com/files/ftp_upload/54453/54453fig1large.jpg" target="_blank">Por favor clique aqui para ver uma versão maior desta figura.</a> <ol start="3"><li> Sob condições de iluminação de espectro total (iluminação cujo espectro se estende para o UV-A gama), isolar um parceiro de acasalamento potencial em cada compartimento final (compartimento escolha) do aquário teste. Garantir que os potenciais parceiros de acasalamento se parecem entre si, tanto quanto possível, em tamanho, coloração e exibir características, de modo que a principal diferença entre os dois parceiros de acasalamento isoladas é se eles aparecem atrás de um filtro que bloqueia ou transmite a luz UV. NOTA: Pode ser preferível para gravar reflectância no espectro visível para permitir a melhor correspondência de cores possível, alembora aqui correspondente foi feito a olho nu. </li><li> Aclimatar indivíduos sob isolamento visuais (filtros opacos) por 15 min. Assegure-se que + e UV - filtros estão no lugar neste momento. Na sequência de aclimatação, remova os filtros opacos. Para assegurar que os desvios laterais não estão presentes (preferências inerente de um dos lados do tanque, independentemente do objecto), atribua aleatoriamente a + UV e UV - lados do tanque para cada cada ensaio. </li><li> Grave a quantidade de tempo que o indivíduo focal passa dentro de zonas preferenciais específicas (áreas próximas aos compartimentos de escolha, neste caso cerca de dois corpos padrão comprimentos) usando cronômetros. No caso do peixe fêmea, isto é a quantidade de tempo que passa perto de cada um dos dois machos, e vice-versa para peixes focal do sexo masculino. NOTA: O software gravador de eventos também pode ser utilizado para gravação. Quinze minutos ensaios foram utilizados neste experimento e os peixes que não visitar cada zona de preferência pelo menos uma vez foram excluídos dos resultados como sendo sem resposta. </li><li> Registar e analisar o tempo que os peixes passam quase vazio UV + e UV - compartimentos para garantir que não há um viés inerente para um ambiente de iluminação especial que podem influenciar a escolha de acasalamento dos peixes. Use testes t pareados para determinar se os indivíduos preferem uma condição UV particular. </li></ol>

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The authors have nothing to disclose.

Espectrofotometria foi bem sucedida na identificação de marcas UV em P. latipinna. Ambos os sexos de P. latipinna possuem marcações UV ao longo de seus lados. Além disso, alguns machos tinha marcas UV em suas barbatanas dorsais, traços encontrados anteriormente a ser importante nas preferências de acasalamento femininos 7. Recomendamos a utilização de espectrofotometria de UV como um mecanismo para detectar a presença de marcas UV. Mais testes poderia d...

Compreender os sinais e sinais utilizados nas interações sociais animais nos permite compreender a variação fenotípica dentro e entre as espécies. Esta variação desempenha um papel importante nos processos evolutivos, tais como a divergência populacional, a selecção sexual, e especiação. Muitas vezes, porém, os pesquisadores estão limitados a explorar as pistas mais óbvias para sistemas sensoriais humanos, especialmente aqueles dentro dos domínios visuais ou auditivas. Uso de espectrofotometria, no entanto, nos permite expandir nossas investigações para além do espectro visível humana e em comprimentos de onda que podem ser importantes em interações sociais em outras espécies. Em particular, a comunicação de curto alcance proporcionada por luz ultravioleta (UV; 300-400 nm) sensibilidade tem o potencial de ser altamente vantajoso durante a escolha do parceiro 1. Muitos predadores visualmente caça de aves e peixes, por exemplo, não são capazes de detectar a radiação UV. Em sistemas onde os machos mostrar elaboradamentepara fêmeas, esses machos reduziria o risco de predação, mantendo a sua capacidade de atrair parceiros, explorando o espectro UV em vez de desenvolver sinais detectáveis ​​no espectro visível 2,3 .. Se alguém deixa de considerar a possibilidade de que os organismos estão se comunicando uns com os outros usam esses &quot;canais de comunicação privados&quot;, motoristas significativas de comportamento e evolução pode ser desperdiçada. Este protocolo descreve uma investigação sobre o uso de pistas de UV para a escolha do companheiro na sailfin molly, Poecilia latipinna, um peixe polígamos que não conheceu anteriormente capacidade de detectar UV ou utilizar marcações UV. Esta espécie de peixe tem uma proximidade filogenética perto de outros peixes Vivípara sensíveis aos raios UV 4 e há evidências de que microspectroscopic P. latipinna, juntamente com outras espécies tais como P. Molly mexicana e P. formosa, possuem uma classe de cones (células fotorreceptoras responsáveis ​​pela covisão lor), que são mais sensíveis para comprimentos de onda UV 5. Nesta espécie dimorfismo sexual, escolha feminina tem desempenhado um papel importante na evolução da dos machos barbatanas coloridas e ampliadas brilhantemente 6-9. Esta metodologia permite-nos explorar se UV é um meio adicional pelo qual as fêmeas avaliar a qualidade do sexo masculino. A detecção e medição de marcações UV sobre P. latipinna usando um espectrofotómetro com uma sonda de fibra óptica é detalhado aqui. Além disso, se mollies fêmeas receptivas diferencialmente associar com machos vistos através de um filtro óptico de transmissão de luz de espectro total, incluindo UV-A ([UV +]; 320-700 nm) e os machos vistos através de um filtro anti-UV ([UV]; 400 - 700 nm) é discutida. Este método tem amplas aplicações para descobrir padrões de sensibilidade UV e cor em peixes e outros organismos, permitindo a investigação sobre uma variedade de questões que envolvem UV e seu papel no comportamento.

<table border="0" cellpadding="0" cellspacing="0" width="1044">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:357px;">Spectrophotometer, P1000</td>
			<td style="width:217px;">Ocean Optics</td>
			<td style="width:119px;"></td>
			<td style="width:351px;">newer models are availabe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:357px;">DT 1000 xenon UV light source</td>
			<td style="width:217px;">Ocean Optics</td>
			<td style="width:119px;"></td>
			<td>newer models are availabe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ocean Optics Overture Software</td>
			<td style="width:217px;">Ocean Optics</td>
			<td style="width:119px;"></td>
			<td>newer software is available</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">R200-Angle-UV bifurcated fiber-optic probe (Guided Wave)</td>
			<td style="width:217px;">Ocean Optics</td>
			<td style="width:119px;"></td>
			<td>newer models are availabe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Certified reflectance standard, white</td>
			<td style="width:217px;">labsphere</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:357px;">75.7 L Aquarium, divided</td>
			<td style="width:217px;">Experimental Builder</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:357px;">Full Spectrum Bulb</td>
			<td style="width:217px;">Nature&#39;s Sunlight</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:357px;">UV blocking sheet</td>
			<td style="width:217px;">GAM UV Sheet</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

assays to detect uv-reflecting structures and determine their importance in mate preference using the sailfin molly poecilia latipinna

Muitos organismos usam pistas e sinais para além sensibilidade humana durante as interações sociais. É importante levar em conta a forma como os organismos percebem seus mundos ao tentar compreender o seu comportamento e ecologia. Sensibilidade ao espectro de ultravioleta (UV; 300-400 nm) é encontrado através de vários géneros de aves, peixes, répteis, anfíbios e até mamíferos. Este protocolo descreve uma técnica pela análise de organismos para a presença de estruturas de reflexão de UV e um método para testar se estes sinais são utilizados como sinais de sociais no contexto da escolha de parceiros. Um espectrofotómetro é utilizado para detectar a presença de reflectância de UV e a variação na intensidade reflexiva entre os indivíduos e os sexos. Um exemplo desta técnica é apresentada em que um teste de escolha do parceiro dicotómica expõe indivíduos sexualmente receptiva a indivíduos do sexo oposto cuja aparência visual pode ser manipulado por meio de filtros que transmitem, quer do espectro de comprimentos de onda de UV ou bloco completo. estesistema permitiu a determinação que as mulheres, mas não do sexo masculino, mollies sailfin (Poecilia latipinna) estavam usando marcas de UV como parte de suas decisões de acasalamento. Estes tipos de estudos servem para ampliar o conhecimento da gama de organismos que utilizam UV e fornecer insights sobre como UV desempenha um papel em suas vidas.

<html> As Figuras 1 e 2 mostram a preferência do aquário de acasalamento configurar e locais de medição UV para nossos experimentos. Medição da reflectância de UV permitiu a determinação de que a P. latipinna possuem características de UV, em especial ao longo dos lados do seu corpo (Figura 3), para além da variação individual em esses traços. Um...

Watch this Scientific Journal Video about Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna at JoVE.com

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

Este protocolo descreve o uso de espectrofotometria para detectar estruturas de reflexão de raios ultravioleta sobre os organismos (neste exemplo, o sailfin molly Poecilia latipinna) e descreve testes de escolha dicotômicas para peixes que permitem inferências a serem feitas sobre o papel das pistas ultravioleta durante a escolha do parceiro.

Ensaios para detectar estruturas de reflexão de UV e determinar a sua importância na preferência do companheiro usando o Sailfin Molly<em&gt; Poecilia latipinna</em

Many organisms use cues and signals beyond human sensitivity during social interactions. It is important to take into account how organisms perceive their ...

assays-to-detect-uv-reflecting-structures-determine-their-importance

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

6.5K visualizações.  Ohio Wesleyan University.  Este protocolo descreve o uso de espectrofotometria para detectar estruturas de reflexão de raios ultravioleta sobre os organismos (neste exemplo, o sailfin molly Poecilia latipinna) e descreve testes de escolha dicotômicas para peixes que permitem inferências a serem feitas sobre o papel das pistas ultravioleta durante a escolha do parceiro. 

Vídeo: Assays to Detect UV-reflecting Structures and Determine their Importance in Mate Preference using the Sailfin Molly Poecilia latipinna

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer basic questions on how light information is processed and shared between rapid and circadian behaviors. Drosophila larvae display a stereotypical avoidance behavior when exposed to light. To investigate light dependent behaviors comparably simple light-dark preference tests can be applied. In vertebrates and arthropods the neural pathways involved in sensing and processing visual inputs partially overlap with those processing photic circadian information. The fascinating question of how the light sensing system and the circadian system interact to keep behavioral outputs coordinated remains largely unexplored. Drosophila is an impacting biological model to approach these questions, due to a small number of neurons in the brain and the availability of genetic tools for neuronal manipulation. The presented light-dark preference assay allows the investigation of a range of visual behaviors including circadian control of phototaxis.

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

Here we describe a light-dark preference test for Drosophila larva. This assay provides information about innate and circadian regulation of light sensing and processing photobehavior.

Luz Ensaio Preferências para estudar inata e circadiano Photobehavior Regulamentado<em&gt; Drosophila</em&gt; As larvas

Light acts as environmental signal to control animal behavior at various levels. The Drosophila larval nervous system is used as a unique model to answer ...

Neurociência

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of the dendritic tree, which eventually leads to neuronal output of action potentials at the axon. The influence of diverse spatial and temporal parameters of specific synaptic input on neuronal output is currently under investigation, e.g. the distance-dependent attenuation of dendritic inputs, the location-dependent interaction of spatially segregated inputs, the influence of GABAergig inhibition on excitatory integration, linear and non-linear integration modes, and many more.
With fast micro-iontophoresis of glutamate and GABA it is possible to precisely investigate the spatial and temporal integration of glutamatergic excitation and GABAergic inhibition. Critical technical requirements are either a triggered fluorescent lamp, light-emitting diode (LED), or a two-photon scanning microscope to visualize dendritic branches without introducing significant photo-damage of the tissue. Furthermore, it is very important to have a micro-iontophoresis amplifier that allows for fast capacitance compensation of high resistance pipettes. Another crucial point is that no transmitter is involuntarily released by the pipette during the experiment.
Once established, this technique will give reliable and reproducible signals with a high neurotransmitter and location specificity. Compared to glutamate and GABA uncaging, fast iontophoresis allows using both transmitters at the same time but at very distant locations without limitation to the field of view. There are also advantages compared to focal electrical stimulation of axons: with micro-iontophoresis the location of the input site is definitely known and it is sure that only the neurotransmitter of interest is released. However it has to be considered that with micro-iontophoresis only the postsynapse is activated and presynaptic aspects of neurotransmitter release are not resolved. In this article we demonstrate how to set up micro-iontophoresis in brain slice experiments.

In this article we introduce fast micro-iontophoresis of neurotransmitters as a technique to investigate integration of postsynaptic signals with high spatial and temporal precision.

Rápido Micro-iontoforese de glutamato e GABA: uma ferramenta útil para investigar Integração Synaptic

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Isolation and Quantification of Botulinum Neurotoxin From Complex Matrices Using the BoTest Matrix Assays

Accurate detection and quantification of botulinum neurotoxin (BoNT) in complex matrices is required for pharmaceutical, environmental, and food sample testing. Rapid BoNT testing of foodstuffs is needed during outbreak forensics, patient diagnosis, and food safety testing while accurate potency testing is required for BoNT-based drug product manufacturing and patient safety. The widely used mouse bioassay for BoNT testing is highly sensitive but lacks the precision and throughput needed for rapid and routine BoNT testing. Furthermore, the bioassay&#39;s use of animals has resulted in calls by drug product regulatory authorities and animal-rights proponents in the US and abroad to replace the mouse bioassay for BoNT testing. Several in vitro replacement assays have been developed that work well with purified BoNT in simple buffers, but most have not been shown to be applicable to testing in highly complex matrices. Here, a protocol for the detection of BoNT in complex matrices using the BoTest Matrix assays is presented. The assay consists of three parts: The first part involves preparation of the samples for testing, the second part is an immunoprecipitation step using anti-BoNT antibody-coated paramagnetic beads to purify BoNT from the matrix, and the third part quantifies the isolated BoNT&#39;s proteolytic activity using a fluorogenic reporter. The protocol is written for high throughput testing in 96-well plates using both liquid and solid matrices and requires about 2 hr of manual preparation with total assay times of 4-26 hr depending on the sample type, toxin load, and desired sensitivity. Data are presented for BoNT/A testing with phosphate-buffered saline, a drug product, culture supernatant, 2% milk, and fresh tomatoes and includes discussion of critical parameters for assay success.

The BoTest Matrix botulinum neurotoxin (BoNT) detection assays rapidly purify and quantify BoNT from a range of sample matrices.&#160;Here, we present a protocol for the detection and quantification of BoNT from both solid and liquid matrices and demonstrate the assay with BOTOX, tomatoes, and milk.

Isolamento e Quantificação da neurotoxina botulínica a partir de matrizes complexas usando os BoTest Matrix Ensaios

Accurate detection and quantification of botulinum neurotoxin (BoNT) in complex matrices is required for pharmaceutical, environmental, and food sample ...

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model

The ability to multiplex assays in studies of complex cellular mechanisms eliminates the need for repetitive experiments, provides internal controls, and decreases waste in costs and reagents. Here we describe optimization of a multiplex assay to assess apoptosis following a palmitic acid (PA) challenge in an in vitro hypothalamic model, using both fluorescent and luminescent based assays to measure viable cell counts and caspase-3/7 activity in a 96-well microtiter plate format. Following PA challenge, viable cells were determined by a resazurin-based fluorescent assay. Caspase-3/7 activity was then determined using a luminogenic substrate, DEVD, and normalized to cell number. This multiplexing assay is a useful technique for determining change in caspase activity following an apoptotic stimulus, such as saturated fatty acid challenge. The saturated fatty acid PA can increase hypothalamic oxidative stress and apoptosis, indicating the potential importance of assays such as that described here in studying the relationship between saturated fatty acids and neuronal function.

Multiplex assays can provide beneficial information for basic cellular mechanisms&#160;and eliminate waste of reagents and unnecessary repetitive experiments. We describe here a multiplex caspase-3/7 activity assay, using fluorescent- and luminescent-based methods, to determine cell viability in an in vitro hypothalamic model following oxidative challenge with palmitic acid.

Utilização de uma caspase Multiplexing ensaio para determinar a apoptose em um modelo de célula hipotalâmica

The ability to multiplex assays in studies of complex cellular mechanisms eliminates the need for repetitive experiments, provides internal controls, and ...

Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure involves the chronic placement of multiple depth electrodes into regions of the brain typically inaccessible via subdural grid electrode placement. SEEG thus provides a unique opportunity to investigate brain function. In this paper we demonstrate how SEEG can be used to investigate the role of the dorsal anterior cingulate cortex (dACC) in cognitive control. We include a description of the SEEG procedure, demonstrating the surgical placement of the electrodes. We describe the components and process required to record local field potential (LFP) data from consenting subjects while they are engaged in a behavioral task. In the example provided, subjects play a cognitive interference task, and we demonstrate how signals are recorded and analyzed from electrodes in the dorsal anterior cingulate cortex, an area intimately involved in decision-making. We conclude with further suggestions of ways in which this method can be used for investigating human cognitive processes.

Stereotactic Electroencephalography (SEEG) is an operative technique used in epilepsy surgery to help localize seizure foci. It also affords a unique opportunity to investigate brain function. Here we describe how SEEG can be used to investigate cognitive processes in human subjects.

Investigar a função do Deep Cortical e estruturas subcorticais Usando estereotáxica Eletroencefalografia: Lições da Cortex cingulado anterior

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure ...

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house single or multiple neurons inside, are considered to make the major contribution in responding to the chemical stimuli. By directly recording action potential in the olfactory sensillum after exposure to stimuli, single sensillum recording (SSR) technique provides a powerful approach for investigating the neural responses of insects to chemical stimuli. For the bed bug, which is a notorious human parasite, multiple types of olfactory sensillum have been characterized. In this study, we demonstrated neural responses of bed bug olfactory sensilla to two chemical stimuli and the dose-dependent responses to one of them using the SSR method. This approach enables researchers to conduct early screening for individual chemical stimuli on the bed bug olfactory sensilla, which would provide valuable information for the development of new bed bug attractants or repellents and benefits the bed bug control efforts.

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

Usando Individual Sensillum Gravação para detectar respostas Olfativo Neurônio de percevejos para Semioquímicos

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

Taste Preference Assay for Adult Drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

Ensaio de Preferência gosto para Adulto<em&gt; Drosophila</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

Enumeration of Neural Stem Cells Using Clonal Assays

Neural stem cells (NSCs) have the ability to self-renew and generate the three major neural lineages &#8212; astrocytes, neurons and oligodendrocytes. NSCs and neural progenitors (NPs) are commonly cultured in vitro as neurospheres. This protocol describes in detail how to determine the NSC frequency in a given cell population under clonal conditions. The protocol begins with the seeding of the cells at a density that allows for the generation of clonal neurospheres. The neurospheres are then transferred to chambered coverslips and differentiated under clonal conditions in conditioned medium, which maximizes the differentiation potential of the neurospheres. Finally, the NSC frequency is calculated based on neurosphere formation and multipotency capabilities. Utilities of this protocol include the evaluation of candidate NSC markers, purification of NSCs, and the ability to distinguish NSCs from NPs. This method takes 13 days to perform, which is much shorter than current methods to enumerate NSC frequency.

Neural stem cells (NSCs) refer to cells&#160;which can self-renew and differentiate into the three neural lineages. Here, we describe a protocol to determine NSC frequency in a given cell population using neurosphere formation and differentiation under clonal conditions.

Enumeração de células-tronco neurais Usando clonais Ensaios

Neural stem cells (NSCs) have the ability to self-renew and generate the three major neural lineages &#8212; astrocytes, neurons and oligodendrocytes. ...

Microglia as a Surrogate Biosensor to Determine Nanoparticle Neurotoxicity

Nanoparticles found in air pollutants can alter neurotransmitter profiles, increase neuroinflammation, and alter brain function. Therefore, the assay described here will aid in elucidating the role of microglia in neuroinflammation and neurodegenerative diseases. The use of microglia, resident immune cells of the brain, as a surrogate biosensor provides novel insight into how inflammatory responses mediate neuronal insults. Here, we utilize an immortalized murine microglial cell line, designated BV2, and describe a method for nanoparticle exposure using silver nanoparticles (AgNPs) as a standard. We describe how to expose microglia to nanoparticles, how to remove nanoparticles from supernatant, and how to use supernatant from activated microglia to determine toxicity, using hypothalamic cell survival as a measure. Following AgNP exposure, BV2 microglial activation was validated using a tumor necrosis factor alpha (TNF-&#945;) enzyme linked immunosorbent assay (ELISA). The supernatant was filtered to remove the AgNP and to allow cytokines and other secreted factors to remain in the conditioned media. Hypothalamic cells were then exposed to supernatant from AgNP activated microglia and survival of neurons was determined using a resazurin-based fluorescent assay. This technique is useful for utilizing microglia as a surrogate biomarker of neuroinflammation and determining the effect of neuroinflammation on other cell types.

Microglia (immune cells of the brain), are used as a surrogate biosensor to determine how nanoparticles influence neurotoxicity. We describe a series of experiments designed to assay microglial response to nanoparticles and exposure of hypothalamic neurons to supernatant from activated microglia to determine neurotoxicity.

Microglia como um Biosensor Surrogate determinar nanopartículas neurotoxicidade

Nanoparticles found in air pollutants can alter neurotransmitter profiles, increase neuroinflammation, and alter brain function. Therefore, the assay ...

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this challenging. Because the ventricles are negative spaces located deep within the brain, the only way to understand their anatomy is by appreciating their boundaries formed by related structures. Looking at a 2D representation of these spaces, in any of the cardinal planes, will not enable visualisation of all of the structures that form the boundaries of the ventricles. Thus, using 2D sections alone requires students to compute their own mental image of the 3D ventricular spaces. The aim of this study was to develop a reproducible method for dissecting the human brain to create an educational resource to enhance student understanding of the intricate relationships between the ventricles and periventricular structures. To achieve this, we created a video resource that features a step-by-step guide using a fiber dissection method to reveal the lateral and third ventricles together with the closely related limbic system and basal ganglia structures. One of the advantages of this method is that it enables delineation of the white matter tracts that are difficult to distinguish using other dissection techniques. This video is accompanied by a written protocol that provides a systematic description of the process to aid in the reproduction of the brain dissection. This package offers a valuable anatomy teaching resource for educators and students alike. By following these instructions educators can create teaching resources and students can be guided to produce their own brain dissection as a hands-on practical activity. We recommend that this video guide be incorporated into neuroanatomy teaching to enhance student understanding of the morphology and clinical relevance of the ventricles.

This paper demonstrates the effective use of a fiber dissection method to reveal the superficial white matter tracts and periventricular structures of the human brain, in three-dimensional space, to aid student comprehension of ventricular morphology.

Explorando o espaço profundo - descobrindo a anatomia das estruturas periventriculares para revelar os ventrículos laterais do cérebro humano

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

Ensaios para detectar estruturas de reflexão de UV e determinar a sua importância na preferência do companheiro usando o Sailfin Molly

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