Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging
Autor: | Claudia L. Treviño, Omar José, Paulina Torres-Rodríguez, Marcela B. Treviño, Ana A. Sánchez-Tusie, Yoloxochitl Sánchez-Guevara, Esperanza Mata-Martínez, Alejandra Solís-López |
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Rok vydání: | 2013 |
Předmět: |
education.field_of_study
medicine.diagnostic_test General Immunology and Microbiology General Chemical Engineering General Neuroscience Population Acrosome reaction Motility Biology Sperm General Biochemistry Genetics and Molecular Biology Flow cytometry Cell biology Single-cell analysis Capacitation medicine education Intracellular |
Zdroj: | Journal of Visualized Experiments. |
ISSN: | 1940-087X |
Popis: | Spermatozoa are male reproductive cells especially designed to reach, recognize and fuse with the egg. To perform these tasks, sperm cells must be prepared to face a constantly changing environment and to overcome several physical barriers. Being in essence transcriptionally and translationally silent, these motile cells rely profoundly on diverse signaling mechanisms to orient themselves and swim in a directed fashion, and to contend with challenging environmental conditions during their journey to find the egg. In particular, Ca(2+)-mediated signaling is pivotal for several sperm functions: activation of motility, capacitation (a complex process that prepares sperm for the acrosome reaction) and the acrosome reaction (an exocytotic event that allows sperm-egg fusion). The use of fluorescent dyes to track intracellular fluctuations of this ion is of remarkable importance due to their ease of application, sensitivity, and versatility of detection. Using one single dye-loading protocol we utilize four different fluorometric techniques to monitor sperm Ca(2+) dynamics. Each technique provides distinct information that enables spatial and/or temporal resolution, generating data both at single cell and cell population levels. |
Databáze: | OpenAIRE |
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