Human Sperm Head Vacuoles Are Related to Nuclear-Envelope Invaginations

Nuclear vacuoles are specific structures present on the head of the human sperm of fertile and non-fertile men. Human sperm head vacuoles have been previously studied using motile sperm organelle morphology examination (MSOME) and their origin related to ab-normal morphology, abnormal chromatin condensation and DNA fragmentation. How-ever, other evidences argued that human sperm vacuoles are physiological structures and consequently, to date, the nature and origin of the nuclear vacuoles remains to be elucidat-ed. Here, we aim to define the incidence, position, morphology, and molecular content of the human sperm vacuoles using transmission electron microscopy (TEM) and immuno-cytochemistry techniques. Results showed that ~50% of the analyzed human sperm cells (n = 1908; 17 normozoospermic human donors) contained vacuoles mainly located (80%) in the anterior head region. A significant positive correlation was also found between the sperm vacuole and nucleus areas. Furthermore, it was confirmed that nuclear vacuoles were invaginations of nuclear envelope containing cytoskeletal proteins and a cytoplas-mic enzyme, discarding a nuclear or acrosomal origin. According to our findings, these human sperm head vacuoles are cellular structures which take the origin from nuclear invaginations and contain perinuclear theca (PT) components, allowing to define a new term of ‘nuclear invaginations’ rather than ‘nuclear vacuoles.