MRNA expression levels of Slc6a19 and Slc1a5 between the LBW and HBW piglets declined gradually from days 0 to 21 of age. No differences in mRNA expression level of Slc6a19 were observed on days 14 and 21 of age, as well as of Slc1a5 on day 21 of age. Age6BW interaction effects were observed for both Slc6a19 and Slc1a5 mRNA expression (P,0.001; Fig. 1). The protein abundances of both B0AT1 and ASCT2 were different from the mRNA expression levels. The protein expression of B0AT1 and ASCT2 was declined from days 0 to 21 of age (P,0.001). Compared with the HBW piglets, the LBW piglets had a lower (P,0.05) protein abundance of B0AT1 on days 0 and 7, as well as of ASCT2 on day 7 of age. No statistical differences inAntibody ASCT2 B0AT1 b-actinCompany Santa Cruz, CA, USA Santa Cruz, CA, USA Santa Cruz, CA, USACatalog GDC-0152 Number Dilution sc130963 sc160811 sc47778 1:500 1:1000 1:doi:10.1371/journal.pone.0050921.tNeutral Amino Acids in Mini-PigletsTable 4. Plasma contents (mmol/L) of neutral amino acids in Huanjiang mini-piglets with HBW1 and LBW2.ItemDay of age 0 HBW LBW 582.bcP-value7 HBW 813.c14 LBW 642.a21 LBW 395.4 190.5 688.3 410.9 141.7 284.2 45.8 88.4 170.3 78.3 112.7 457.d dHBW 358.8 198.1d 739.4 454.9 137.9 250.3 38.2 74.6 150.2 77.0 113.4 484.dHBW 380.6 167.5 782.9 452.2 153.8 303.3 49.d dLBW 377.8 165.5d 731.2 465.0 145.6 297.7 46.4 99.8 179.1 81.4 130.3 449.dSEM 97.69 36.91 33.41 24.43 7.13 16.03 7.74 7.54 7.49 5.13 8.47 12.Age 0.330 0.001 0.136 0.693 ,0.001 0.079 0.009 0.144 0.484 0.712 0.124 0.BW 0.362 0.038 0.113 0.009 0.276 0.415 0.015 0.836 0.504 0.827 0.266 0.Age6BW 0.829 0.046 0.660 0.776 0.393 0.568 0.037 0.385 0.736 0.943 0.449 0.Thr Ser Gly Ala Cys Val Met Ile Leu Tyr Phe HMPL-013 site Proa-d 1757.9 347.8 624.4 446.7 151.9 406.0 140.7 78.2 239.8 114.0 171.2 506.a279.7 539.5 421.7 138.1 370.6 96.4 54.1 238.8 110.8 151.5 495.b474.c1127.8 674.1 148.9 355.4 81.bc a776.9b147.1 359.9 62.cd115.9 217.0 111.0 151.4 465.140.3 216.8 122.6 139.4 433.123.9 193.4 81.8 118.3 431.Values within a row without a common superscript letter differ (P,0.05). HBW, high birth weight; LBW, low birth weight. doi:10.1371/journal.pone.0050921.tprotein abundances of B0AT1 and ASCT2 were observed on days 14 and 21 of age. There were interaction between age and BW on both Slc6a19 and Slc1a5 protein expression(P,0.001; Fig. 2).DiscussionThis study investigated the NAA contents of plasma, liver, and skeletal muscle, as well as jejunal expression profiles of their transporters in suckling Huanjiang mini-piglets with HBW or LBW. The novel and important findings from this study are thatthe LBW piglets had alterations in the contents of NAA in plasma (including Ser, Ala and Met), liver (including Thr, Ser, Gly, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe and Pro) and muscle (including Gly) during the early sucking period, which were associated with expression changes of their intestinal transporters at both mRNA and protein levels, with a lower expression level of Slc6a19 (B0AT1) and Slc1a5 (ASCT2) 1379592 in the LBW piglets. There were age6BW interaction effects on plasma (including Ser and Met), liver (including Thr, Ser, Gly, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe and Pro) and muscle (including Gly) contents of NAA, as wellTable 5. Liver contents ( ) of neutral amino acids in Huanjiang mini-piglets with HBW1 and LBW 2.ItemDay of age 0 HBW LBW 0.45 0.44 0.57 0.57 0.45 0.67 0.25 0.49 0.87 0.40 0.69 0.d d b b ab c c c c d d cP-value7 HBW 0.54 0.52 0.64 0.68 0.43 0.82 0.27 0.60 1.08 0.47 0.MRNA expression levels of Slc6a19 and Slc1a5 between the LBW and HBW piglets declined gradually from days 0 to 21 of age. No differences in mRNA expression level of Slc6a19 were observed on days 14 and 21 of age, as well as of Slc1a5 on day 21 of age. Age6BW interaction effects were observed for both Slc6a19 and Slc1a5 mRNA expression (P,0.001; Fig. 1). The protein abundances of both B0AT1 and ASCT2 were different from the mRNA expression levels. The protein expression of B0AT1 and ASCT2 was declined from days 0 to 21 of age (P,0.001). Compared with the HBW piglets, the LBW piglets had a lower (P,0.05) protein abundance of B0AT1 on days 0 and 7, as well as of ASCT2 on day 7 of age. No statistical differences inAntibody ASCT2 B0AT1 b-actinCompany Santa Cruz, CA, USA Santa Cruz, CA, USA Santa Cruz, CA, USACatalog Number Dilution sc130963 sc160811 sc47778 1:500 1:1000 1:doi:10.1371/journal.pone.0050921.tNeutral Amino Acids in Mini-PigletsTable 4. Plasma contents (mmol/L) of neutral amino acids in Huanjiang mini-piglets with HBW1 and LBW2.ItemDay of age 0 HBW LBW 582.bcP-value7 HBW 813.c14 LBW 642.a21 LBW 395.4 190.5 688.3 410.9 141.7 284.2 45.8 88.4 170.3 78.3 112.7 457.d dHBW 358.8 198.1d 739.4 454.9 137.9 250.3 38.2 74.6 150.2 77.0 113.4 484.dHBW 380.6 167.5 782.9 452.2 153.8 303.3 49.d dLBW 377.8 165.5d 731.2 465.0 145.6 297.7 46.4 99.8 179.1 81.4 130.3 449.dSEM 97.69 36.91 33.41 24.43 7.13 16.03 7.74 7.54 7.49 5.13 8.47 12.Age 0.330 0.001 0.136 0.693 ,0.001 0.079 0.009 0.144 0.484 0.712 0.124 0.BW 0.362 0.038 0.113 0.009 0.276 0.415 0.015 0.836 0.504 0.827 0.266 0.Age6BW 0.829 0.046 0.660 0.776 0.393 0.568 0.037 0.385 0.736 0.943 0.449 0.Thr Ser Gly Ala Cys Val Met Ile Leu Tyr Phe Proa-d 1757.9 347.8 624.4 446.7 151.9 406.0 140.7 78.2 239.8 114.0 171.2 506.a279.7 539.5 421.7 138.1 370.6 96.4 54.1 238.8 110.8 151.5 495.b474.c1127.8 674.1 148.9 355.4 81.bc a776.9b147.1 359.9 62.cd115.9 217.0 111.0 151.4 465.140.3 216.8 122.6 139.4 433.123.9 193.4 81.8 118.3 431.Values within a row without a common superscript letter differ (P,0.05). HBW, high birth weight; LBW, low birth weight. doi:10.1371/journal.pone.0050921.tprotein abundances of B0AT1 and ASCT2 were observed on days 14 and 21 of age. There were interaction between age and BW on both Slc6a19 and Slc1a5 protein expression(P,0.001; Fig. 2).DiscussionThis study investigated the NAA contents of plasma, liver, and skeletal muscle, as well as jejunal expression profiles of their transporters in suckling Huanjiang mini-piglets with HBW or LBW. The novel and important findings from this study are thatthe LBW piglets had alterations in the contents of NAA in plasma (including Ser, Ala and Met), liver (including Thr, Ser, Gly, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe and Pro) and muscle (including Gly) during the early sucking period, which were associated with expression changes of their intestinal transporters at both mRNA and protein levels, with a lower expression level of Slc6a19 (B0AT1) and Slc1a5 (ASCT2) 1379592 in the LBW piglets. There were age6BW interaction effects on plasma (including Ser and Met), liver (including Thr, Ser, Gly, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe and Pro) and muscle (including Gly) contents of NAA, as wellTable 5. Liver contents ( ) of neutral amino acids in Huanjiang mini-piglets with HBW1 and LBW 2.ItemDay of age 0 HBW LBW 0.45 0.44 0.57 0.57 0.45 0.67 0.25 0.49 0.87 0.40 0.69 0.d d b b ab c c c c d d cP-value7 HBW 0.54 0.52 0.64 0.68 0.43 0.82 0.27 0.60 1.08 0.47 0.
