Midkine Promotes Proliferation of Primordial Germ Cells by Inhibiting the Expression of the Deleted in Azoospermia-Like Gene

Abstract

Primordial germ cell (PGC) development is an area of research that is hampered by low cell numbers as well as difficulty in isolation. They are, however, required for the production of gametes and as such represent an important area of understanding that has widespread implications for fertility and reproductive technologies. Here we investigated the role of the heparin-binding growth factor midkine (MK) on PGC development, first using our established model of porcine stem cell-derived PGC-like cells and then confirming our findings in PGC. Our results show that MK has a mitogenic effect on PGC, mediated through an increased cell proliferation as well as decreased apoptosis. Upon further investigation, we found these effects concomitant with the decreased expression of the germ cell-specific gene deleted in azoospermia-like (DAZL). This decrease in DAZL expression, and consequent decreases in the meiosis-related genes SCP3 and DMC1, suggest a role for MK in preventing a shift in the PGC phenotype toward meiosis. MK instead increases activity of mitotic pathways in PGC, keeping them in a proliferative, less differentiated state. Lentiviral-mediated overexpression of DAZL further confirmed its role in promoting meiosis in and reducing proliferation of PGC. These effects were mitigated by the addition of MK, which was able to limit the effect of this DAZL overexpression. Furthermore, a loss-of-function study showed that a DAZL knockdown by small interfering RNA had the same effect as that induced by the addition of MK. Taken together, these data suggest that MK is able to maintain a proliferative PGC phenotype mediated by the suppression of DAZL in early germ cells.