Molecular Constraints of Sperm Sex Sorting via TLR7/8 Activation

In modern livestock, the demand for sperm sex selection technologies is high, as the ability to deliberately produce offspring of a specific sex offers significant economic advantages. Traditionally, sperm sorting is performed using Fluorescence-Activated Cell Sorting. However, the flow cytometric method is expensive, technically complex, and associated with reduced sperm viability. An alternative promising method involves the use of Toll-like TLR7/8 receptors for the selective binding of spermatozoa of a particular sex. It was discovered previously that the activation of TLR7/8 by its ligand(s) selectively inhibits the motility of X-bearing sperm without affecting the motility of Y-bearing sperm. The swim-up technique, which separates sperm based on sex chromosome type by isolating fractions enriched in either X- or Y-bearing gametes due to differences in their motility, can be used with this method. Sperm sex sorting via the TLR7/8 activation is cheap, technically non-complex, and does not affect sperm viability negatively. The goal of this review is to provide an overview of the TLR7/8-dependent sperm sorting method. Further, we discuss why the method of sperm sorting via TLR7/8 activation is successfully implemented in some animal species (such as murine, caprine, ovine, and bovine) but fails in others, like swine and canine.