Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Membrane proteomic analysis reveals the intestinal development is deteriorated by intrauterine growth restriction in piglets


The alterations of the intestinal proteome were observed in intrauterine growth restriction (IUGR) piglets during early life by gel-based approaches. Nevertheless, how IUGR affects the intestinal membrane proteome during neonatal development remains unclear. Here, we applied the iTRAQ-based proteomics technology and biochemical analysis to investigate the impact of IUGR on the membrane proteome of the jejunal mucosa in the piglets. Three hundred sixty-one membrane proteins were screened by functional prediction. Among them, eight, five, and one differentially expressed membrane proteins were identified between IUGR and NBW piglets at day 0, day 7, and day 21 after birth, respectively. Differentially expressed membrane proteins (DEMPs) including F1SBL3, F1RRW8, F1S539, F1S2Z2, F1RIR2, F1RUF2 I3LP60, Q2EN79, and F1SIH8 were reduced while the relative abundance of I3L6A2, F1SCJ1, F1RI18, I3LRJ7, and F1RNN0 were increased in IUGR piglets than NBW piglets. From the aspects of function, F1RRW8, F1S539, F1S2Z2, and F1RIR2 are mainly associated with D2 dopamine receptor binding, transmembrane transport of small molecules, signal transduction, and translocation of GLUT4, respectively, and F1SIH8, I3LRJ7, and F1RNN0 are related to autophagy, metabolism of vitamins, and intracellular protein transport. Additionally, IUGR decreased the level of proteins (F1RRW8, Q2EN79, and F1RI18) that are involved in response to oxidative stress.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9


  1. Archer SL (2013) Mitochondrial dynamics--mitochondrial fission and fusion in human diseases. N Engl J Med 369:2236–2251.

  2. Band AM, Kuismanen E (2005) Localization of plasma membrane t-SNAREs syntaxin 2 and 3 in intracellular compartments. BMC Cell Biol 6:26.

  3. Barel O, Shorer Z, Flusser H, Ofir R, Narkis G, Finer G, Shalev H, Nasasra A, Saada A, Birk OS (2008) Mitochondrial complex III deficiency associated with a homozygous mutation in UQCRQ. Am J Hum Genet 82:1211–1216.

  4. Baserga M, Bertolotto C, Maclennan NK, Hsu JL, Pham T, Laksana GS, Lane RH (2004) Uteroplacental insufficiency decreases small intestine growth and alters apoptotic homeostasis in term intrauterine growth retarded rats. Early Hum Dev 79:93–105.

  5. Boumil RM, Letts VA, Roberts MC, Christine L, Mahaffey CL, Zhong-Wei Z, Tobias M, Frankel WN (2010) A missense mutation in a highly conserved alternate exon of dynamin-1 causes epilepsy in fitful mice. PLoS Genet 6:182–188.

  6. Bozzetti V, Tagliabue PE, Visser GH, van Bel F, Gazzolo D (2013) Feeding issues in IUGR preterm infants. Early Hum Dev 89(Suppl 2):S21–S23.

  7. Codogno P, Meijer AJ (2005) Autophagy and signaling: their role in cell survival and cell death. Cell Death Differ 12(Suppl 2):1509–1518.

  8. D’Inca R, Che L, Thymann T, Sangild PT, Huërou-Luron IL (2010a) Intrauterine growth restriction reduces intestinal structure and modifies the response to colostrum in preterm and term piglets. Livest Sci 133:20–22.

  9. D’Inca R, Kloareg M, Gras-Le Guen C, Le Huerou-Luron I (2010b) Intrauterine growth restriction modifies the developmental pattern of intestinal structure, transcriptomic profile, and bacterial colonization in neonatal pigs. J Nutr 140:925–931.

  10. D’Inca R, Gras-Le Guen C, Che L, Sangild PT, Le Huerou-Luron I (2011) Intrauterine growth restriction delays feeding-induced gut adaptation in term newborn pigs. Neonatology 99:208–216.

  11. Fan F, Funk L, Lou X (2016) Dynamin 1- and 3-mediated endocytosis is essential for the development of a large central synapse in vivo journal of neuroscience the official. J Soc Neurosci 36:6097–6115.

  12. Ferguson SM, Brasnjo G, Hayashi M, Wolfel M, Collesi C, Giovedi S, Raimondi A, Gong LW, Ariel P, Paradise S, O’Toole E, Flavell R, Cremona O, Miesenbock G, Ryan TA, De Camilli P (2007) A selective activity-dependent requirement for dynamin 1 in synaptic vesicle endocytosis. Science 316:570–574.

  13. Fujimaki A, Watanabe K, Mori T, Kimura C, Shinohara K, Wakatsuki A (2011) Placental oxidative DNA damage and its repair in preeclamptic women with fetal growth restriction. Placenta 32:367–372.

  14. Fujita K, Nakamura Y, Oka T, Ito H, Tamura T, Tagawa K, Sasabe T, Katsuta A, Motoki K, Shiwaku H, Sone M, Yoshida C, Katsuno M, Eishi Y, Murata M, Taylor JP, Wanker EE, Kono K, Tashiro S, Sobue G, La Spada AR, Okazawa H (2013) A functional deficiency of TERA/VCP/p97 contributes to impaired DNA repair in multiple polyglutamine diseases. Nat Commun 4:1816.

  15. Glover LE, Lee JS, Colgan SP (2016) Oxygen metabolism and barrier regulation in the intestinal mucosa. J Clin Invest 126:3680–3688.

  16. Haq S, Grondin J, Banskota S, Khan WI (2019) Autophagy: roles in intestinal mucosal homeostasis and inflammation. J Biomed Sci 26:19.

  17. Hsueh W, Caplan MS, Qu XW, Tan XD, De Plaen IG, Gonzalez-Crussi F (2003) Neonatal necrotizing enterocolitis: clinical considerations and pathogenetic concepts. Pediatr Dev Pathol 6:6–23.

  18. Huang JB, Liu YL, Sun PW, Lv XD, Du M, Fan XM (2010) Molecular mechanisms of congenital heart disease. Cardiovasc Pathol 19:e183–e193.

  19. Huang S, Li N, Liu C, Li T, Wang W, Jiang L, Li Z, Han D, Tao S, Wang J (2019a) Characteristics of the gut microbiota colonization, inflammatory profile, and plasma metabolome in intrauterine growth restricted piglets during the first 12 hours after birth. J Microbiol 57:748–758.

  20. Huang S, Wu Z, Liu C, Han D, Feng C, Wang S, Wang J (2019b) Milk fat globule membrane supplementation promotes neonatal growth and alleviates inflammation in low-birth-weight mice treated with lipopolysaccharide. Biomed Res Int 2019:4876078–4876010.

  21. Ji Y, Dai Z, Sun S, Ma X, Yang Y, Tso P, Wu G, Wu Z (2018) Hydroxyproline attenuates dextran sulfate sodium-induced colitis in mice: involvment of the NF-κB signaling and oxidative stress. Mol Nutr Food Res e1800494.

  22. Ju JS, Fuentealba RA, Miller SE, Jackson E, Piwnica-Worms D, Baloh RH, Weihl CC (2009) Valosin-containing protein (VCP) is required for autophagy and is disrupted in VCP disease. J Cell Biol 187:875–888.

  23. Kittler R, Putz G, Pelletier L, Poser I, Heninger AK, Drechsel D, Fischer S, Konstantinova I, Habermann B, Grabner H, Yaspo ML, Himmelbauer H, Korn B, Neugebauer K, Pisabarro MT, Buchholz F (2004) An endoribonuclease-prepared siRNA screen in human cells identifies genes essential for cell division. Nature 432:1036–1040.

  24. Li N, Wang W, Wu G, Wang J (2017a) Nutritional support for low birth weight infants: insights from animal studies. Br J Nutr 117:1390–1402.

  25. Li X, Jiang X, Xu X, Zhu C, Yi W (2017b) Imaging of protein-specific glycosylation by glycan metabolic tagging and in situ proximity ligation. Carbohydr Res 448:148–154.

  26. Li N, Huang S, Jiang L, Dai Z, Li T, Han D, Wang J (2019) Characterization of the early life microbiota development and predominant Lactobacillus species at distinct gut segments of low- and normal-birth-weight piglets. Front Microbiol 10:797.

  27. Ma J, Chen T, Wu S, Yang C, Bai M, Shu K, Li K, Zhang G, Jin Z, He F, Hermjakob H, Zhu Y (2019) iProX: an integrated proteome resource. Nucleic Acids Res 47:1211–1217.

  28. Maeda A, Maeda T, Golczak M, Palczewski K (2008) Retinopathy in mice induced by disrupted all-trans-retinal clearance. J Biol Chem 283:26684–26693.

  29. McNally KE, Faulkner R, Steinberg F, Gallon M, Ghai R, Pim D, Langton P, Pearson N, Danson CM, Nagele H, Morris LL, Singla A, Overlee BL, Heesom KJ, Sessions R, Banks L, Collins BM, Berger I, Billadeau DD, Burstein E, Cullen PJ (2017) Retriever is a multiprotein complex for retromer-independent endosomal cargo recycling. Nat Cell Biol 19:1214–1225.

  30. Meerang M, Ritz D, Paliwal S, Garajova Z, Bosshard M, Mailand N, Janscak P, Hubscher U, Meyer H, Ramadan K (2011) The ubiquitin-selective segregase VCP/p97 orchestrates the response to DNA double-strand breaks. Nat Cell Biol 13:1376–1382.

  31. Miller SL, Huppi PS, Mallard C (2016) The consequences of fetal growth restriction on brain structure and neurodevelopmental outcome. J Physiol 594:807–823.

  32. Mittal M, Siddiqui MR, Tran K, Reddy SP, Malik AB (2014) Reactive oxygen species in inflammation and tissue injury. Antioxid Redox Signal 20:1126–1167.

  33. Nemoto Y, Kumagai T, Ishizawa K, Miura Y, Shiraishi T, Morimoto C, Sakai K, Omizo H, Yamazaki O, Tamura Y, Fujigaki Y, Kawachi H, Kuro OM, Uchida S, Shibata S (2019) Phosphate binding by sucroferric oxyhydroxide ameliorates renal injury in the remnant kidney model. Sci Rep 9:1732.

  34. Nishiyama Y, Kataoka T, Yamato K, Taguchi T, Yamaoka K (2012) Suppression of dextran sulfate sodium-induced colitis in mice by radon inhalation. Mediat Inflamm 2012:239617–239611.

  35. Ott M, Gogvadze V, Orrenius S, Zhivotovsky B (2007) Mitochondria, oxidative stress and cell death. Apoptosis 12:913–922.

  36. Regev RH, Reichman B (2004) Prematurity and intrauterine growth retardation--double jeopardy? Clin Perinatol 31:453–473.

  37. Sangild PT, Siggers RH, Schmidt M, Elnif J, Bjornvad CR, Thymann T, Grondahl ML, Hansen AK, Jensen SK, Boye M, Moelbak L, Buddington RK, Westrom BR, Holst JJ, Burrin DG (2006) Diet- and colonization-dependent intestinal dysfunction predisposes to necrotizing enterocolitis in preterm pigs. Gastroenterology 130:1776–1792.

  38. Tomasz D, Daniel G, Rafal R, Agnieszka S, Jaroslaw S, Romuald Z, Ryszard O (2007) Urinary excretion rates of 8-oxoGua and 8-oxodG and antioxidant vitamins level as a measure of oxidative status in healthy, full-term newborns. Free Radic Res 41:997-1004.

  39. Tonks NK (2006) Protein tyrosine phosphatases: from genes, to function, to disease. Nat Rev Mol Cell Biol 7:833–846.

  40. Tsybovsky Y, Molday RS, Palczewski K (2010) The ATP-binding cassette transporter ABCA4: structural and functional properties and role in retinal disease. Adv Exp Med Biol 703:105–125.

  41. Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9:799–809.

  42. Wang X, Qiao S, Yin Y, Yue L, Wang Z, Wu G (2007) A deficiency or excess of dietary threonine reduces protein synthesis in jejunum and skeletal muscle of young pigs. J Nutr 137:1442–1446.

  43. Wang J, Chen L, Li D, Yin Y, Wang X, Li P, Dangott LJ, Hu W, Wu G (2008) Intrauterine growth restriction affects the proteomes of the small intestine, liver, and skeletal muscle in newborn pigs. J Nutr 138:60–66.

  44. Wang X, Wu W, Lin G, Li D, Wu G, Wang J (2010) Temporal proteomic analysis reveals continuous impairment of intestinal development in neonatal piglets with intrauterine growth restriction. J Proteome Res 9:924–935.

  45. Wang X, Lin G, Liu C, Feng C, Zhou H, Wang T, Li D, Wu G, Wang J (2014) Temporal proteomic analysis reveals defects in small-intestinal development of porcine fetuses with intrauterine growth restriction. J Nutr Biochem 25:785–795.

  46. Wang W, Degroote J, Van Ginneken C, Van Poucke M, Vergauwen H, Dam TM, Vanrompay D, Peelman LJ, De Smet S, Michiels J (2016) Intrauterine growth restriction in neonatal piglets affects small intestinal mucosal permeability and mRNA expression of redox-sensitive genes. FASEB J 30:863–873.

  47. Wang J, Feng C, Liu T, Shi M, Wu G, Bazer FW (2017) Physiological alterations associated with intrauterine growth restriction in fetal pigs: causes and insights for nutritional optimization. Mol Reprod Dev 84:897–904.

  48. Wang X, Zhu Y, Feng C, Lin G, Wu G, Li D, Wang J (2018) Innate differences and colostrum-induced alterations of jejunal mucosal proteins in piglets with intra-uterine growth restriction. Br J Nutr 119:734–747.

  49. Watts GD, Wymer J, Kovach MJ, Mehta SG, Mumm S, Darvish D, Pestronk A, Whyte MP, Kimonis VE (2004) Inclusion body myopathy associated with Paget disease of bone and frontotemporal dementia is caused by mutant valosin-containing protein. Nat Genet 36:377–381.

  50. Woodman PG (2003) p97, a protein coping with multiple identities. J Cell Sci 116:4283–4290.

  51. Wu G, Bazer FW, Wallace JM, Spencer TE (2006) Board-invited review: intrauterine growth retardation: implications for the animal sciences. J Anim Sci 84:2316–2337.

  52. Yatsenko AN, Wiszniewski W, Zaremba CM, Jamrich M, Lupski JR (2005) Evolution of ABCA4 proteins in vertebrates. J Mol Evol 60:72–80.

  53. Ye Y, Meyer HH, Rapoport TA (2001) The AAA ATPase Cdc48/p97 and its partners transport proteins from the ER into the cytosol. Nature 414:652–656.

  54. Yoo M, Kim BG, Lee SJ, Jeong HJ, Park JW, Seo DW, Kim YK, Lee HY, Han JW, Kang JS, Bae GU (2015) Syntaxin 4 regulates the surface localization of a promyogenic receptor Cdo thereby promoting myogenic differentiation. Skelet Muscle 5:28.

  55. Yu Y, Zhao Y, Teng F, Li J, Guan Y, Xu J, Lv X, Guan F, Zhang M, Chen L (2018) Berberine improves cognitive deficiency and muscular dysfunction via activation of the AMPK/SIRT1/PGC-1a pathway in skeletal muscle from naturally aging rats. J Nutr Health Aging 22:710–717.

  56. Zhang L, Ashendel CL, Becker GW, Morre DJ (1994) Isolation and characterization of the principal ATPase associated with transitional endoplasmic reticulum of rat liver. J Cell Biol 127:1871–1883.

  57. Zhong X, Shen Y, Ballar P, Apostolou A, Agami R, Fang S (2004) AAA ATPase p97/valosin-containing protein interacts with gp78, a ubiquitin ligase for endoplasmic reticulum-associated degradation. J Biol Chem 279:45676–45684.

  58. Zhu YH, Lin G, Dai ZL, Zhou TJ, Yuan TL, Feng CP, Chen F, Wu GY, Wang JJ (2015) Developmental changes in polyamines and autophagic marker levels in normal and growth-restricted fetal pigs. J Anim Sci 93:3503–3511.

Download references


We thank the Mianyang New-hope Livestock Farming Co. Ltd. in Sichuan Province, China, for their assistance in this study. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium ( via the iProX partner repository (Ma et al. 2019) with the dataset identifier PXD013315.


This work was supported by the National Natural Science Foundation of China (31630074), the Beijing Municipal Natural Science Foundation (S170001), the National Key Research and Development Program of China (2016YFD0500506, 2018YDF0501002), the 111 Project (B16044), and Jinxinnong Animal Science Developmental Foundation.

Author information

Correspondence to Junjun Wang.

Ethics declarations

All experiments were carried out with the approval of China Agricultural University Animal Care and Use Committee (CAU20170114–1, Beijing, China).

Competing interests

The authors declared that they have no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOCX 16 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Huang, S., Liu, C., Li, N. et al. Membrane proteomic analysis reveals the intestinal development is deteriorated by intrauterine growth restriction in piglets. Funct Integr Genomics 20, 277–291 (2020).

Download citation


  • iTRAQ
  • Intrauterine growth restriction
  • Neonatal piglets
  • Membrane proteome
  • Intestine