Abstract
Different approaches can be used to assess sperm function in different conditions, i.e. sperm storage, freezing-thawing or activation by induction of capacitation and acrosome reaction. In this review we will focus on the assays routinely performed in our laboratories, giving a literature support to critically analyse different approaches. In fact, researchers usually tend to look for the âone shotâ parameter that could explain itself a specific process; it is our conviction that a multiparametric approach is still more valid, as some changes in sperm function are very complex and could be explained only by operating in different ways. Sperm motility, the most evident sperm characteristic, should be assessed by computer-aided sperm analysers that permit an objective evaluation of the motility and its kinematic parameters. Commercial and open source instruments are available and could be profitably used together with specific statistical approaches. The use of microscopy, and particularly fluorescent microscopy, could be a very useful tool to assess different parameters in sperm cells both by fluorophores that give indication of a determined function, and by immunolocalization of proteins, that permits the discover of new features or to explain particular sperm functions. The same substrates could be used also in flow cytometry: the difference is that it permits to study wider sperm populations (and their sub-population distribution). Flow cytometry is undergoing a very wide use in spermatology and technical and experimental rigor is needed to obtain reliable results. Metabolic assessment of sperm features, particularly energetic supply, ATP formation and other enzyme activities, could represent a very important challenge to acquire new information and complete/integrate those derived from other techniques. Finally, functional assays such as oocyte binding and in vitro fertilization, represent a very strong tool to assess sperm function in vitro, as they could evidence the functional intactness of some pathways.
Highlights
Spermatozoa are cells programmed to deliver male genetic material to the female gamete, permitting fertilization and born of a new individual.To be able to achieve this goal, spermatozoa should undergo numerous functional adaptations from the time they are ejaculated, passing through male genital tract, female one, to reach the fertilization site.In vitro assays to assess different sperm functions are, at present, a very important tool to understand and discover the pathways implicated in sperm function and their changes
Analysis of protein expression is of fundamental importance in trying to investigate sperm function, as new proteins could be discovered in the mature sperm cell and their function could be elucidated
The use of fluorochrome-bound antibodies against specific proteins or activated proteins (i.e.phosphorilated ones) is of actual importance (Piehler et al, 2006); it should be noted that, in the case of flow cytometry, the instrument output when using antibodies could be related to the presence/absence of the determined protein and, partially, on the intensity of the signal
Summary
Spermatozoa are cells programmed to deliver male genetic material to the female gamete, permitting fertilization and born of a new individual. To be able to achieve this goal, spermatozoa should undergo numerous functional adaptations from the time they are ejaculated, passing through male genital tract, female one, to reach the fertilization site (usually the ampulla of the oviduct; Suarez and Pacey, 2006). In vitro assays to assess different sperm functions are, at present, a very important tool to understand and discover the pathways implicated in sperm function and their changes. The present review was aimed at presenting some of the most important approaches in mammalian sperm function analysis, with particular focus on our laboratory experience and at furnishing a critical view of the different parameters examined
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