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Electroantennogram and Single Sensillum Recording in Insect Antennae

Electrophysiology is an invaluable technique to quickly and quantitatively assess the response of the olfactory system to odor stimuli. For measuring the response of the insect antenna, two basic techniques exist, electroantennography and single sensillum recording. Here, we de ...

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A Protocol for Heterologous Expression and Functional Assay for Mouse Pheromone Receptors

Innate social behaviors like intermale aggression, fear, and mating rituals are important for survival and propagation of a species. In mice, these behaviors have been implicated to be mediated by peptide pheromones that are sensed by a class of G protein-coupled receptors, vomeronasal re ...

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Identification of Chemosensory Receptor Genes from Vertebrate Genomes

Chemical senses are essential for the survival of animals. In vertebrates, mainly three different types of receptors, olfactory receptors (ORs), vomeronasal receptors type 1 (V1Rs), and vomeronasal receptors type 2 (V2Rs), are responsible for the detection of chemicals in the environ ...

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Genetic Manipulation to Analyze Pheromone Responses: Knockouts of Multiple Receptor Genes

Gene targeting in the mouse is an essential technique to study gene function in vivo. Multigene families encoding vomeronasal receptor (VR) type 1 and type 2 consist of ~300 intact genes, which are clustered at multiple loci in the mouse genome. To understand the function of VRs and neurons expressi ...

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Analysis of Ascarosides from Caenorhabditis elegans Using Mass Spectrometry and NMR Spectroscopy

The nematode Caenorhabditis elegans secretes a family of water-soluble small molecules, known as the ascarosides, into its environment and uses these ascarosides in chemical communication. The ascarosides are derivatives of the 3,6-dideoxysugar ascarylose, modified with di ...

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Chemical Analysis of Aquatic Pheromones in Fish

Pheromones are chemicals that pass between members of the same species that have inherent meaning. In the case of fish, pheromones are water-soluble and found in low concentrations. As such, sensitive and selective methods are needed to separate and analyze these pheromones from an environ ...

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Murine Nonvolatile Pheromones: Isolation of Exocrine-Gland Secreting Peptide 1

Our search for a substance recognized by the vomeronasal neurons revealed that the extra-orbital lacrimal gland (ELG) isolated from adult male mice produced the male-specific peptide pheromone exocrine gland-secreting peptide 1 (ESP1). The following protocol reveals how ESP1 may be ...

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Pheromones in the Fruit Fly

The identification of pheromones (chemical communication cues) is critical to our understanding of complex social behavior in insects and other animals. In this chapter, we describe analytical methods for the purification of lipid pheromones by thin layer chromatography and the qu ...

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Female Moth Pheromones

The sex pheromone, a volatile secreted by a female moth, is stored in the pheromone gland and can be easily extracted with hexane. The extract is effectively analyzed using a gas chromatography combined with an electro-antennogram detector (GC-EAD) and a mass spectrometry (GC-MS), both of which ...

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Using Somatic-Cell Nuclear Transfer to Study Aging

In mammals, a diploid genome following fertilization of haploid cells, an egg, and a spermatozoon is unique and irreproducible. This implies that the generated unique diploid genome is doomed with the individual’s inevitable demise. Since it was first reported in 1997 that Dolly the sheep had b ...

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The Use of Calorie Restriction Mimetics to Study Aging

Calorie restriction (CR) has a variety of effects on extending lifespan and delaying the onset of age-related diseases, and it is accepted as the only established experimental antiaging intervention. Several pharmacological agents that can replicate the beneficial effects of CR, ca ...

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Aging Studies in Drosophila Melanogaster

Drosophila is a genetically tractable system ideal for investigating the mechanisms of aging and developing interventions for promoting healthy aging. Here we describe methods commonly used in Drosophila aging research. These include basic approaches for preparation of diets a ...

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Methods for Creating Mutations in C. elegans That Extend Lifespan

The principle of commonly used methods to create mutations in the nematode Caenorhabditis elegans (C. elegans) is straightforward. In general, worms are exposed to a dose of mutagen resulting in DNA damages and mutations. Screening the progeny of the mutagenized animals for a certain pheno ...

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Studying the Replicative Life Span of Yeast Cells

The budding yeast Saccharomyces cerevisiae is a useful model for elucidating the pathways that control life span and the influence of environmental factors, such as calorie restriction (CR). For 75 years, CR has been studied for its ability to delay diseases of aging in mammals, from cancer to car ...

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Cell Sorting of Young and Senescent Cells

Cellular senescence is the irreversible loss of proliferative potential and is accompanied by a number of phenotypic changes. First described by Hayflick and Moorhead in 1961, it has since become a popular model to study cellular aging. The replicative lifespan of human fibroblasts is het ...

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Analysis of Biomarkers of Caloric Restriction in Aging Cells

Caloric restriction (CR) has been extensively documented for its profound role in effectively extending maximum lifespan in many different species. However, the accurate mechanisms, especially at the cellular level, for CR-induced aging delay are still under intense investigat ...

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Digital Image Analysis of Cells Stained with the Senescence-Associated -Galactosidase Assay

Cellular senescence plays important roles in the aging process of complex organisms, in response to stress and in tumor suppression. Several markers can be used to identify senescent cells, of which the most widely used is the senescence-associated β-galactosidase (SABG) activity. Here ...

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Single-Neuron Transcriptome and Methylome Sequencing for Epigenomic Analysis of Aging

Enormous heterogeneity in transcription and signaling is the feature that slows down progress in our understanding of the mechanisms of normal aging and age-related diseases. This is critical for neurobiology of aging where the enormous diversity of neuronal populations presents a s ...

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Epigenetic Biomarker to Determine Replicative Senescence of Cultured Cells

Somatic cells change continuously during culture expansion—long-term culture evokes increasing cell size, declining differentiation potential, and ultimate cell cycle arrest upon senescence. These changes are of particular relevance for cellular therapy which necess ...

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Cell Senescence Culturing Methods

Development of therapeutic approaches that slow or ablate the adverse physiological and pathological changes associated with aging has been considered as an important goal for gerontological research. As cellular senescence is characterized as the basis for aging in organisms, c ...

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