Sperm Intracellular Calcium Evaluation
Adriana Casao1, Sara Miguel-Jimenez2, Rosaura Pérez-Pe2
1Grupo BIOFITER, Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Veterinaria, Instituto Universitario de Investigación en Ciencias Ambientales de Aragón, Universidad de Zaragoza, Zaragoza, Spain. adriana@unizar.es.
Abstract:
The evaluation of the intracellular calcium is essential in studying sperm capacitation and acrosome reaction, among other processes. Here, we describe the use of chlortetracycline (CTC) to study Ca2+ distribution in relation to the sperm capacitation state, and the evaluation of Ca2+ levels in spermatozoa using a low or a high-affinity Ca2+ dye (Rhod-5 N-AM and Fluo-4-AM, respectively) by flow cytometry. Rhod-5 N-AM staining method also allows the study of Ca2+ in subcellular compartments, whereas the changes in intracellular Ca2+ levels can be measured in spermatozoa loaded with Fluo-4-AM by plate reader fluorometry.


