Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Development of pre-implantation porcine blastocysts cultured within alginate hydrogel systems either supplemented with secreted phosphoprotein 1 or conjugated with Arg-Gly-Asp Peptide

Taylor D. Laughlin A , Jeremy R. Miles B C , Elane C. Wright-Johnson B , Lea A. Rempel B , Clay A. Lents B and Angela K. Pannier A C
+ Author Affliations
- Author Affliations

A Department of Biological Systems Engineering, University of Nebraska – Lincoln, PO Box 830726, Lincoln, NE 68583, USA.

B USDA1, U.S. Meat Animal Research Center (USMARC), PO Box 166, Clay Center, Nebraska 68933, USA.

C Corresponding authors. Email: jeremy.miles@ars.usda.gov; apannier2@unl.edu

Reproduction, Fertility and Development - https://doi.org/10.1071/RD16366
Submitted: 15 September 2016  Accepted: 14 March 2017   Published online: 28 April 2017

Abstract

Although deficiencies in porcine blastocyst elongation play a significant role in early embryonic mortality and establishment of within-litter developmental variation, the exact mechanisms of elongation are poorly understood. Secreted phosphoprotein 1 (SPP1) is increased within the uterine milieu during early porcine pregnancy and contains an Arg-Gly-Asp (RGD) peptide sequence that binds to cell surface integrins on the uterine endometrium and trophectoderm, promoting cell adhesion and migration. The aim of the present study was to evaluate the development of preimplantation porcine blastocysts encapsulated and cultured within alginate hydrogels either supplemented with SPP1 or conjugated with RGD. Blastocysts encapsulated within alginate hydrogels supplemented with SPP1 or conjugated with RGD had increased survival compared with non-encapsulated control blastocysts. In addition, the percentage of blastocysts encapsulated within RGD hydrogels that underwent morphological changes was greater than that of blastocysts encapsulated within standard alginate hydrogels or SPP1-supplemented hydrogels. Finally, only blastocysts encapsulated within RGD hydrogels had both increased expression of steroidogenic and immune responsiveness transcripts and increased 17β-oestradiol production, consistent with blastocysts undergoing elongation in vivo. These results illustrate the importance of the integrin-binding RGD peptide sequence for stimulating the initiation of blastocyst elongation.

Additional keywords: embryo elongation, embryology, gene expression, oestrogen.


References

Ashworth, C. J., Pickard, A. R., Miller, S. J., Flint, A. P., and Diehl, J. R. (1997). Comparative studies of conceptus–endometrial interactions in Large White × Landrace and Meishan gilts. Reprod. Fertil. Dev. 9, 217–225.
Comparative studies of conceptus–endometrial interactions in Large White × Landrace and Meishan gilts.CrossRef | 1:STN:280:DyaK2szlt1Wmsg%3D%3D&md5=d3e9c85ceba35a8050458c2fa286781aCAS | open url image1

Bailey, D. W., Dunlap, K. A., Erikson, D. W., Patel, A. K., Bazer, F. W., Burghardt, R. C., and Johnson, G. A. (2010). Effects of long-term progesterone exposure on porcine uterine gene expression: progesterone alone does not induce secreted phosphoprotein 1 (osteopontin) in glandular epithelium. Reproduction 140, 595–604.
Effects of long-term progesterone exposure on porcine uterine gene expression: progesterone alone does not induce secreted phosphoprotein 1 (osteopontin) in glandular epithelium.CrossRef | 1:CAS:528:DC%2BC3cXhtlCqurrF&md5=d2433aa0eb3105cb6d9e0bd2b5840b7eCAS | open url image1

Bazer, F. W., Geisert, R. D., Thatcher, W. W., and Roberts, R. M. (1982) The establishment and maintenance of pregnancy. In ‘Control of Pig Reproduction’. (Eds D. J. A. Cole and G. R. Foxcroft.) pp. 227–252. (Butterworth Scientific: London.)

Bazer, F. W., Thatcher, W. W., Martinat-Botte, F., and Terqui, M. (1988). Conceptus development in large white and prolific Chinese Meishan pigs. J. Reprod. Fertil. 84, 37–42.
Conceptus development in large white and prolific Chinese Meishan pigs.CrossRef | 1:STN:280:DyaL1M%2Fjs12muw%3D%3D&md5=9f2bf1319f753a0bc62a09dacd7e21b7CAS | open url image1

Bazer, F. W., Spencer, T. E., Johnson, G. A., Burghardt, R. C., and Wu, G. (2009). Comparative aspects of implantation. Reproduction 138, 195–209.
Comparative aspects of implantation.CrossRef | 1:CAS:528:DC%2BD1MXptlemt74%3D&md5=8c3cc60f5cf5365558bdd25f965af094CAS | open url image1

Bazer, F. W., Kim, J., Ka, H., Johnson, G. A., Wu, G., and Song, G. (2012). Select nutrients in the uterine lumen of sheep and pigs affect conceptus development. J. Reprod. Dev. 58, 180–188.
Select nutrients in the uterine lumen of sheep and pigs affect conceptus development.CrossRef | 1:CAS:528:DC%2BC38XptVKju7k%3D&md5=9421846c25295c9d6198b7178118b840CAS | open url image1

Blomberg, L. A., and Zuelke, K. A. (2005). Expression analysis of the steroidogenic acute regulatory protein (STAR) gene in developing porcine conceptuses. Mol. Reprod. Dev. 72, 419–429.
Expression analysis of the steroidogenic acute regulatory protein (STAR) gene in developing porcine conceptuses.CrossRef | 1:CAS:528:DC%2BD2MXhtFGmurvE&md5=c1b88c4da33f607996e3b7e33e5b4659CAS | open url image1

Blomberg, L. A., Long, E. L., Sonstegard, T. S., Van Tassell, C. P., Dobrinsky, J. R., and Zuelke, K. A. (2005). Serial analysis of gene expression during elongation of the peri-implantation porcine trophectoderm (conceptus). Physiol. Genomics 20, 188–194.
Serial analysis of gene expression during elongation of the peri-implantation porcine trophectoderm (conceptus).CrossRef | 1:CAS:528:DC%2BD2MXitVGjtbs%3D&md5=46b1c0df21cbac5d817ef637f63afe8bCAS | open url image1

Blomberg, L. A., Garrett, W. M., Guillomot, M., Miles, J. R., Sonstegard, T. S., Van Tassell, C. P., and Zuelke, K. A. (2006a). Transcriptome profiling of the tubular porcine conceptus identifies the differential regulation of growth and developmentally associated genes. Mol. Reprod. Dev. 73, 1491–1502.
Transcriptome profiling of the tubular porcine conceptus identifies the differential regulation of growth and developmentally associated genes.CrossRef | 1:CAS:528:DC%2BD28XhtFGhtr7E&md5=628ab83d436497ae67c4a6c74f15c9a8CAS | open url image1

Blomberg, L. A., Miles, J. R., and Zuelke, K. A. (2006b). Unique expression patterns of differentiation, growth, and cell structure factors in the elongating porcine conceptus. Reprod. Fertil. Dev. 18, 170.
Unique expression patterns of differentiation, growth, and cell structure factors in the elongating porcine conceptus.CrossRef | open url image1

Blomberg, L. A., Schreier, L., and Li, R. W. (2010). Characteristics of peri-implantation porcine concepti population and maternal milieu influence the transcriptome profile. Mol. Reprod. Dev. 77, 978–989.
Characteristics of peri-implantation porcine concepti population and maternal milieu influence the transcriptome profile.CrossRef | 1:CAS:528:DC%2BC3cXhsVCktrnI&md5=17342627883a540af7cf68102deeb7b7CAS | open url image1

Corbin, C. J., Berger, T., Ford, J. J., Roselli, C. E., Sienkiewicz, W., Trainor, B. C., Roser, J. F., Vidal, J. D., Harada, N., and Conley, A. J. (2009). Porcine hypothalamic aromatase cytochrome P450: isoform characterization, sex-dependent activity, regional expression, and regulation by enzyme inhibition in neonatal boars. Biol. Reprod. 81, 388–395.
Porcine hypothalamic aromatase cytochrome P450: isoform characterization, sex-dependent activity, regional expression, and regulation by enzyme inhibition in neonatal boars.CrossRef | 1:CAS:528:DC%2BD1MXptVaiur0%3D&md5=f6e24f501a51fc9be5bfc3c045920b33CAS | open url image1

Erikson, D. W., Burghardt, R. C., Bayless, K. J., and Johnson, G. A. (2009). Secreted phosphoprotein 1 (SPP1, osteopontin) binds to integrin alpha v beta 6 on porcine trophectoderm cells and integrin alpha v beta 3 on uterine luminal epithelial cells, and promotes trophectoderm cell adhesion and migration. Biol. Reprod. 81, 814–825.
Secreted phosphoprotein 1 (SPP1, osteopontin) binds to integrin alpha v beta 6 on porcine trophectoderm cells and integrin alpha v beta 3 on uterine luminal epithelial cells, and promotes trophectoderm cell adhesion and migration.CrossRef | 1:CAS:528:DC%2BD1MXhtlWrtr%2FN&md5=7ac68e648b921982e0f93207d34b677aCAS | open url image1

Federation of Animal Science Societies (FASS) (2010). ‘Guide for the Care and Use of Agricultural Animals in Research and Teaching.’ 3rd edn. (FASS: Champaign, IL.)

Foxcroft, G. R., Dixon, W. T., Dyck, M. K., Novak, S., Harding, J. C., and Almeida, F. C. (2009). Prenatal programming of postnatal development in the pig. Soc. Reprod. Fertil. Suppl. 66, 213–231.
| 1:STN:280:DC%2BD1MjgvFaksA%3D%3D&md5=18a27c7414c7a482e7785f857ef4d9dfCAS | open url image1

Garlow, J. E., Ka, H., Johnson, G. A., Burghardt, R. C., Jaeger, L. A., and Bazer, F. W. (2002). Analysis of osteopontin at the maternal–placental interface in pigs. Biol. Reprod. 66, 718–725.
Analysis of osteopontin at the maternal–placental interface in pigs.CrossRef | 1:CAS:528:DC%2BD38XhvVeiur8%3D&md5=0a2b7efa4b4bbe8481ecbfc7321d7ca0CAS | open url image1

Geisert, R. D., and Yelich, J. V. (1997). Regulation of conceptus development and attachment in pigs. J. Reprod. Fertil. Suppl. 52, 133–149.
| 1:STN:280:DyaK1c3msFCgtw%3D%3D&md5=b4f4467e90594bb72fe8693adacab63aCAS | open url image1

Geisert, R. D., Brookbank, J. W., Roberts, R. M., and Bazer, F. W. (1982). Establishment of pregnancy in the pig: II. Cellular remodeling of the porcine blastocyst during elongation on day 12 of pregnancy. Biol. Reprod. 27, 941–955.
Establishment of pregnancy in the pig: II. Cellular remodeling of the porcine blastocyst during elongation on day 12 of pregnancy.CrossRef | 1:STN:280:DyaL3s%2FnsFKjtw%3D%3D&md5=a5d7194e3cb19662a1b6111481dc610aCAS | open url image1

Geisert, R., Fazleabas, A., Lucy, M., and Mathew, D. (2012). Interaction of the conceptus and endometrium to establish pregnancy in mammals: role of interleukin 1beta. Cell Tissue Res. 349, 825–838.
Interaction of the conceptus and endometrium to establish pregnancy in mammals: role of interleukin 1beta.CrossRef | 1:CAS:528:DC%2BC38Xht1yltbrM&md5=8a0b4671aac706bf1f5c83fae9d2015cCAS | open url image1

Geisert, R. D., Lucy, M. C., Whyte, J. J., Ross, J. W., and Mathew, D. J. (2014). Cytokines from the pig conceptus: roles in conceptus development in pigs. J. Anim. Sci. Biotechnol. 5, 51.
Cytokines from the pig conceptus: roles in conceptus development in pigs.CrossRef | open url image1

Jaeger, L. A., Johnson, G. A., Ka, H., Garlow, J. G., Burghardt, R. C., Spencer, T. E., and Bazer, F. W. (2001). Functional analysis of autocrine and paracrine signalling at the uterine–conceptus interface in pigs. Reprod. Suppl. 58, 191–207.
| 1:CAS:528:DC%2BD38XitFent7k%3D&md5=8d0e094c995e946916f55084245f0c59CAS | open url image1

Johnson, G. A., Burghardt, R. C., Bazer, F. W., and Spencer, T. E. (2003). Osteopontin: roles in implantation and placentation. Biol. Reprod. 69, 1458–1471.
Osteopontin: roles in implantation and placentation.CrossRef | 1:CAS:528:DC%2BD3sXosV2kur8%3D&md5=cfc397026bb119b05e0afce49521ff09CAS | open url image1

Livak, K. J., and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(–Delta Delta C(T)) method. Methods 25, 402–408.
Analysis of relative gene expression data using real-time quantitative PCR and the 2(–Delta Delta C(T)) method.CrossRef | 1:CAS:528:DC%2BD38XhtFelt7s%3D&md5=27001fc4d390214adec9d2e6245c9fd6CAS | open url image1

Mickos, H., Bahr, J., and Luning, B. (1990). The three-dimensional structure of the RGD (Arg-Gly-Asp) adhesion sequence of fibronectin; NMR studies of the peptides RGDS and GRGDSP. Acta Chem. Scand. 44, 161–164.
The three-dimensional structure of the RGD (Arg-Gly-Asp) adhesion sequence of fibronectin; NMR studies of the peptides RGDS and GRGDSP.CrossRef | 1:CAS:528:DyaK3cXitFaltL4%3D&md5=3a2d287f61ef768f2698487716e91efaCAS | open url image1

Miles, J. R., Blomberg, L. A., Krisher, R. L., Everts, R. E., Sonstegard, T. S., Van Tassell, C. P., and Zuelke, K. A. (2008a). Comparative transcriptome analysis of in vivo- and in vitro-produced porcine blastocysts by small amplified RNA–serial analysis of gene expression (SAR-SAGE). Mol. Reprod. Dev. 75, 976–988.
Comparative transcriptome analysis of in vivo- and in vitro-produced porcine blastocysts by small amplified RNA–serial analysis of gene expression (SAR-SAGE).CrossRef | 1:CAS:528:DC%2BD1cXlsVWjtro%3D&md5=75b7f9e54fe946b9655fee1495e2c720CAS | open url image1

Miles, J. R., Freking, B. A., Blomberg, L. A., Vallet, J. L., and Zuelke, K. A. (2008b). Conceptus development during blastocyst elongation in lines of pigs selected for increased uterine capacity or ovulation rate. J. Anim. Sci. 86, 2126–2134.
Conceptus development during blastocyst elongation in lines of pigs selected for increased uterine capacity or ovulation rate.CrossRef | 1:CAS:528:DC%2BD1cXhtFansLfF&md5=fa5b453ba62bcf8a6beb8a7e7e103113CAS | open url image1

Miles, J. R., Vallet, J. L., Freking, B. A., and Nonneman, D. J. (2009). Molecular cloning and characterisation of heparanase mRNA in the porcine placenta throughout gestation. Reprod. Fertil. Dev. 21, 757–772.
Molecular cloning and characterisation of heparanase mRNA in the porcine placenta throughout gestation.CrossRef | 1:CAS:528:DC%2BD1MXnvVOhuro%3D&md5=3d34aff12707cd28bb6d2cf340359ab5CAS | open url image1

Pope, W. F. (1994). Embryonic mortality in swine. In ‘Embryonic Mortality in Domestic Species’. (Eds M. T. Zavy and R. D. Geisert.) pp. 53–77. (CRC Press Inc.: Boca Raton, FL.)

Pope, W. F., and First, N. L. (1985). Factors affecting the survival of pig embryos. Theriogenology 23, 91–105.
Factors affecting the survival of pig embryos.CrossRef | open url image1

Redmer, D. A., and Day, B. N. (1981). Ovarian activity and hormonal patterns in gilts fed allyl trenbolone. J. Anim. Sci. 53, 1088–1094.
Ovarian activity and hormonal patterns in gilts fed allyl trenbolone.CrossRef | 1:CAS:528:DyaL3MXmtVanurc%3D&md5=be24a758bf621925dd593093028e470bCAS | open url image1

Rowley, J. A., Madlambayan, G., and Mooney, D. J. (1999). Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials 20, 45–53.
Alginate hydrogels as synthetic extracellular matrix materials.CrossRef | 1:CAS:528:DyaK1MXjtlWnug%3D%3D&md5=f4799199c7723b07d37ecbb225e54bdaCAS | open url image1

Sargus-Patino, C. N., Wright, E. C., Plautz, S. A., Miles, J. R., Vallet, J. L., and Pannier, A. K. (2014). In vitro development of preimplantation porcine embryos using alginate hydrogels as a three-dimensional extracellular matrix. Reprod. Fertil. Dev. 26, 943–953.
In vitro development of preimplantation porcine embryos using alginate hydrogels as a three-dimensional extracellular matrix.CrossRef | 1:CAS:528:DC%2BC2cXhsVClsLvM&md5=c37385ef2a08702c09a2bef4e9e00121CAS | open url image1

Steel, R. G. D., Torrie, J. H., and Dickey, D. A. (1997) ‘Principles and Procedures of Statistics: A Biometrical Approach.’ 3rd edn. (McGraw-Hill Inc.: New York, NY.)

Stice, S. L., Robl, J. M., Ponce de Leon, F. A., Jerry, J., Golueke, P. G., Cibelli, J. B., and Kane, J. J. (1998). Cloning: new breakthroughs leading to commercial opportunities. Theriogenology 49, 129–138.
Cloning: new breakthroughs leading to commercial opportunities.CrossRef | 1:CAS:528:DyaK1cXnsFCruw%3D%3D&md5=0eb930580aa0278386102bf1840c7f0aCAS | open url image1

Stocco, D. M., and Clark, B. J. (1997). The role of the steroidogenic acute regulatory protein in steroidogenesis. Steroids 62, 29–36.
The role of the steroidogenic acute regulatory protein in steroidogenesis.CrossRef | 1:CAS:528:DyaK2sXhtVylt7Y%3D&md5=306ba056f6b0b26e5e5fd093a98e63cfCAS | open url image1

Valdez Magaña, G., Rodríguez, A., Zhang, H., Webb, R., and Alberio, R. (2014). Paracrine effects of embryo-derived FGF4 and BMP4 during pig trophoblast elongation. Dev. Biol. 387, 15–27.
Paracrine effects of embryo-derived FGF4 and BMP4 during pig trophoblast elongation.CrossRef | open url image1

Vejlsted, M., Du, Y., Vajta, G., and Maddox-Hyttel, P. (2006). Post-hatching development of the porcine and bovine embryo-defining criteria for expected development in vivo and in vitro. Theriogenology 65, 153–165.
Post-hatching development of the porcine and bovine embryo-defining criteria for expected development in vivo and in vitro.CrossRef | open url image1

Waclawik, A. (2011). Novel insights into the mechanisms of pregnancy establishment: regulation of prostaglandin synthesis and signaling in the pig. Reproduction 142, 389–399.
Novel insights into the mechanisms of pregnancy establishment: regulation of prostaglandin synthesis and signaling in the pig.CrossRef | 1:CAS:528:DC%2BC3MXht1Cjur3J&md5=03046adfb00246dd20ee2589e98c3522CAS | open url image1

Yelich, J. V., Pomp, D., and Geisert, R. D. (1997). Ontogeny of elongation and gene expression in the early developing porcine conceptus. Biol. Reprod. 57, 1256–1265.
Ontogeny of elongation and gene expression in the early developing porcine conceptus.CrossRef | 1:CAS:528:DyaK2sXmvFSkur4%3D&md5=2633ba6536ba75666044dd1a93ce856eCAS | open url image1



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