Methodology A rapid protocol for purification of total RNA for tissues collected from pigs at a slaughterhouse

Mendez, Victor Mechan; Avelar, Ernesto; Morales, Adriana; Cervantes, M.; Araiza, Alfonso; González, Deyarina

doi:10.4238/2011.december.22.3

Cited by 32 publications

(24 citation statements)

References 5 publications

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“…The samples of intestinal mucosa were treated to extract total RNA by pulverization into liquid N, and homogenized into Trizol reagent (Invitrogen, Carlsbad, CA, USA) as reported by Méndez et al. (). Purified RNA was eluted with 30 μ l of RNase‐free distilled water and stored at −82 °C.…”

Section: Methodsmentioning

confidence: 99%

Dietary levels of protein and free amino acids affect pancreatic proteases activities, amino acids transporters expression and serum amino acid concentrations in starter pigs

Morales

Buenabad

Castillo

et al. 2016

Animal Physiology Nutrition

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The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA-supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat-soybean meal diet were measured in two 14-d trials. The LPAA diet contained free L-Lys, L-Thr, DL-Met, L-Leu, L-Ile, L-Val, L-His, L-Trp and L-Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b AT in duodenum, B AT1 in ileum (p < 0.05), and tended to express more y LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.

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Section: Methodsmentioning

confidence: 99%

Dietary levels of protein and free amino acids affect pancreatic proteases activities, amino acids transporters expression and serum amino acid concentrations in starter pigs

Morales

Buenabad

Castillo

et al. 2016

Animal Physiology Nutrition

Self Cite

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“…Samples of jejunal mucosa, liver, LM and SM were treated to extract total RNA by the Trizol reagent (Invitrogen, Corp., Carlsbad, CA, USA), as reported by Méndez et al. (). Purified RNA was then eluted with 30 μ l of RNase‐free water and stored at −82 °C.…”

Section: Methodsmentioning

confidence: 99%

Effect of L‐lysine on expression of selected genes, serum concentration of amino acids, muscle growth and performance of growing pigs

Morales

García

Arce

et al. 2014

Animal Physiology Nutrition

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Lysine (Lys) is the first limiting amino acid (AA) in most feed formulations for pigs and most abundant, along with leucine, in muscle proteins. An experiment was conducted with 17 pigs (17.7 ± 0.05 kg initial BW) to identify a role of dietary Lys in the control of protein synthesis in pigs. Fourteen pigs were randomly assigned to one of the two wheat-based dietary treatments: Lys-deficient, 3.0 g/kg (DEF) and Lys-adequate, 10.8 g/kg (ADE). Samples from jejunum mucosa, liver, Longissumus and Semitendinosus muscles, and blood were collected. The other three pigs were sacrificed at the beginning of the trial to measure basal carcass composition. Weight gain, gain:feed ratio, Lys intake and loin eye area were greater in ADE than in DEF pigs (p < 0.01). Muscle-related carcass characteristics were better, and myosin heavy chain IIb expression (MyHC IIb) in Semitendinosus was higher in ADE than in DEF pigs. Expression of AA transporters CAT-1 was lower (p < 0.05), serum Lys was higher and serum Val was lower in pigs fed the ADE diet. The higher muscularity, MyHC IIb expression in Semitendinosus muscle and Lys serum of pigs fed the ADE diet suggest that Lys increases growth rate not only by functioning as protein construction unit but also as potential control of the protein synthesis process.

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“…Samples of the jejunum mucosa, liver, LD, and ST were treated to extract total RNA using the Trizol reagent (Invitrogen, Carlsbad, CA, USA) as reported by Méndez et al (2011). Purified RNA was then eluted with 30 μL RNase-free water and stored at -82°C.…”

Section: Total Rna Extraction and Purificationmentioning

confidence: 99%

Gene expression, serum amino acid levels, and growth performance of pigs fed dietary leucine and lysine at different ratios

García¹,

Morales²,

Araiza³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. We examined 96 pigs (28.1 ± 0.83 kg) to analyze the effect of Leu:Lys ratios on expression of the cationic amino acid transporters b 0,+ and CAT-1 in the jejunum and liver as well as myosin expression in 2 muscles to estimate the optimum standardized ileal digestible (SID) Leu:Lys ratio for growth rate and efficiency. A wheatand wheat bran-based diets were formulated to meet the requirements of SID amino acids other than Leu (0.70%) and Lys (0.80%). L-Leu was added to the basal diet in 5 SID Leu:Lys ratios (88, 100, 120, 140, and 160% in diets 1-5). Tissue samples were collected from 8 pigs with ratios of 88, 120, and 160%. Relative expression of b 0,+ , CAT-1, and myosin was analyzed. b 0,+ expression in the jejunum was higher but lower in the liver of pigs with the 120% ratio compared to those with the 88 or 160% ratio; myosin expression in longissimus dorsi was also higher in pigs with the 120% ratio (P < 0.05). CAT-1 was lower in the jejunum and longissimus dorsi of pigs with 120 or 160% ratios than in pigs with 88%. Serum concentration of nearly all amino acids decreased with excess dietary Leu (P < 0.05). The SID Leu:Lys of 104 and 109% 1590 H. García et al. ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (1): 1589-1601 (2015 optimized average daily gain and feed conversion ratio, respectively. Thus, the dietary Leu:Lys ratio affects the expression of genes coding for amino acid transporters and myosin, the availability of Lys, and the growth rate and efficiency in pigs.

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Methodology A rapid protocol for purification of total RNA for tissues collected from pigs at a slaughterhouse

Cited by 32 publications

References 5 publications

Dietary levels of protein and free amino acids affect pancreatic proteases activities, amino acids transporters expression and serum amino acid concentrations in starter pigs

Dietary levels of protein and free amino acids affect pancreatic proteases activities, amino acids transporters expression and serum amino acid concentrations in starter pigs

Effect of L‐lysine on expression of selected genes, serum concentration of amino acids, muscle growth and performance of growing pigs

Gene expression, serum amino acid levels, and growth performance of pigs fed dietary leucine and lysine at different ratios

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