Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/BAT1 Is Required for Optimal Preimplantation Embryo Development in Mice

L-proline (Pro) has previously been shown to support normal development of mouse embryos. Recently we have shown that Pro improves subsequent embryo development when added to fertilisation medium during in vitro fertilisation of mouse oocytes. The mechanisms by which Pro improves embryo development are still being elucidated but likely involve signalling pathways that have been observed in Pro-mediated differentiation of mouse embryonic stem cells. In this study, we show that BAT1, a neutral amino acid transporter that accepts Pro, is expressed in mouse preimplantation embryos, along with the accessory protein ACE2. BAT1 knockout () mice have decreased fertility, in terms of litter size and preimplantation embryo development in vitro. In embryos from wild-type (WT) mice, excess unlabelled Pro inhibited radiolabelled Pro uptake in oocytes and 4-8-cell stage embryos. Radiolabelled Pro uptake was reduced in 4-8-cell stage embryos, but not in oocytes, from mice compared to those from WT mice. Other BAT1 substrates, such as alanine and leucine, reduced uptake of Pro in WT but not in BAT1 knockout embryos. Addition of Pro to culture medium improved embryo development. In WT embryos, Pro increased development to the cavitation stage (on day 4); whereas in BAT1 knockout embryos Pro improved development to the 5-8-cell (day 3) and blastocyst stages (day 6) but not at cavitation (day 4), suggesting BAT1 is the main contributor to Pro uptake on day 4 of development. Our results highlight transporter redundancy in the preimplantation embryo.