Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 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...
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MDPI AG
2022-12-01
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| author | Tamara Treleaven Matthew Zada Rajini Nagarajah Charles G. Bailey John E. J. Rasko Michael B. Morris Margot L. Day |
| author_facet | Tamara Treleaven Matthew Zada Rajini Nagarajah Charles G. Bailey John E. J. Rasko Michael B. Morris Margot L. Day |
| author_sort | Tamara Treleaven |
| collection | DOAJ |
| description | 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 B<sup>0</sup>AT1, a neutral amino acid transporter that accepts Pro, is expressed in mouse preimplantation embryos, along with the accessory protein ACE2. B<sup>0</sup>AT1 knockout (<i>Slc6a19<sup>−/−</sup></i>) 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 <i>Slc6a19<sup>−/−</sup></i> mice compared to those from WT mice. Other B<sup>0</sup>AT1 substrates, such as alanine and leucine, reduced uptake of Pro in WT but not in B<sup>0</sup>AT1 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 B<sup>0</sup>AT1 knockout embryos Pro improved development to the 5–8-cell (day 3) and blastocyst stages (day 6) but not at cavitation (day 4), suggesting B<sup>0</sup>AT1 is the main contributor to Pro uptake on day 4 of development. Our results highlight transporter redundancy in the preimplantation embryo. |
| first_indexed | 2024-03-11T10:05:49Z |
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| last_indexed | 2024-03-11T10:05:49Z |
| publishDate | 2022-12-01 |
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| spelling | doaj.art-5d18cc2b9ed44b3ea03a67edc9551eda2023-11-16T15:04:58ZengMDPI AGCells2073-44092022-12-011211810.3390/cells12010018Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in MiceTamara Treleaven0Matthew Zada1Rajini Nagarajah2Charles G. Bailey3John E. J. Rasko4Michael B. Morris5Margot L. Day6School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, AustraliaSchool of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, AustraliaGene & Stem Cell Therapy Program Centenary Institute, The University of Sydney, Camperdown, NSW 2050, AustraliaGene & Stem Cell Therapy Program Centenary Institute, The University of Sydney, Camperdown, NSW 2050, AustraliaGene & Stem Cell Therapy Program Centenary Institute, The University of Sydney, Camperdown, NSW 2050, AustraliaSchool of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, AustraliaSchool of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, AustraliaL-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 B<sup>0</sup>AT1, a neutral amino acid transporter that accepts Pro, is expressed in mouse preimplantation embryos, along with the accessory protein ACE2. B<sup>0</sup>AT1 knockout (<i>Slc6a19<sup>−/−</sup></i>) 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 <i>Slc6a19<sup>−/−</sup></i> mice compared to those from WT mice. Other B<sup>0</sup>AT1 substrates, such as alanine and leucine, reduced uptake of Pro in WT but not in B<sup>0</sup>AT1 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 B<sup>0</sup>AT1 knockout embryos Pro improved development to the 5–8-cell (day 3) and blastocyst stages (day 6) but not at cavitation (day 4), suggesting B<sup>0</sup>AT1 is the main contributor to Pro uptake on day 4 of development. Our results highlight transporter redundancy in the preimplantation embryo.https://www.mdpi.com/2073-4409/12/1/18prolineamino acid transporter<i>Slc6a19</i>B<sup>0</sup>AT1preimplantation embryomouse |
| spellingShingle | Tamara Treleaven Matthew Zada Rajini Nagarajah Charles G. Bailey John E. J. Rasko Michael B. Morris Margot L. Day Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice Cells proline amino acid transporter <i>Slc6a19</i> B<sup>0</sup>AT1 preimplantation embryo mouse |
| title | Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice |
| title_full | Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice |
| title_fullStr | Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice |
| title_full_unstemmed | Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice |
| title_short | Stage-Specific L-Proline Uptake by Amino Acid Transporter Slc6a19/B<sup>0</sup>AT1 Is Required for Optimal Preimplantation Embryo Development in Mice |
| title_sort | stage specific l proline uptake by amino acid transporter slc6a19 b sup 0 sup at1 is required for optimal preimplantation embryo development in mice |
| topic | proline amino acid transporter <i>Slc6a19</i> B<sup>0</sup>AT1 preimplantation embryo mouse |
| url | https://www.mdpi.com/2073-4409/12/1/18 |
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