Porcine Growth Research Articles

MEG3 Promotes Differentiation of Porcine Satellite Cells by Sponging miR-423-5p to Relieve Inhibiting Effect on SRF

Although thousands of long noncoding RNAs (lncRNAs) have been identified in porcine growth and development, the regulation mechanisms of functional lncRNAs have not been well explored. In this study, using 5′- and 3′-rapid amplification of cDNA ends (RACE) assays, we obtained two different variants of lncRNA maternally expressed gene 3 (MEG3), namely, MEG3 v1 and MEG3 v2, that were both highly expressed in porcine skeletal muscle and in the early stage of the differentiation of porcine satellite cells. Moreover, we identified the core transcript MEG3 v2. Functional analyses showed that MEG3 overexpression could effectively arrest myoblasts in the G1 phase, inhibit DNA replication, and promote myoblast differentiation, whereas MEG3 knockdown resulted in the opposite effects. Interestingly, the expression of serum response factor (SRF), a crucial transcription factor for myogenesis process, remarkably increased and decreased in mRNA and protein levels with the respective overexpression and knockdown of MEG3. Dual luciferase reporter assay showed that MEG3 could attenuate the decrease of luciferase activity of SRF induced by miR-423-5p in a dose-dependent manner. MEG3 overexpression could relieve the inhibitory effect on SRF and myoblast differentiation induced by miR-423-5p. In addition, results of RNA immunoprecipitation analysis suggested that MEG3 could act as a ceRNA for miR-423-5p. Our findings initially established a novel connection among MEG3, miR-423-5p, and SRF in porcine satellite cell differentiation. This novel role of MEG3 may shed new light on understanding of molecular regulation of lncRNA in porcine myogenesis.

Changes in porcine growth, biochemical parameters, histopathology, and melamine residue levels in tissues induced by consumption of melamine and cyanuric acid

Changes in porcine growth, biochemical parameters, histopathology, and melamine residue levels in tissues induced by consumption of melamine and cyanuric acid

Myeloid zinc finger 1 (MZF1) is the most important transcriptional factor for porcine follistatin promoter

Follistatin (FS) is a secreted protein, which was originally isolated from porcine follicular fluid. Expression of follistatin is tightly regulated during porcine growth and development. To study the essential transcriptional regions of the porcine FS promoter, ten primer pairs were designed to amplify segments with different lengths of the FS promoter from −1 800 to +16 bp. The products were then inserted into the pGL3-basic vector to analyze the relative luciferase activity. The results showed that the most remarkable changes of promoter activity were observed between constructs (−302/+16 bp)-FS and (−180/+16 bp)-FS (P<0.01). Further research showed that the reconstructed reporter plasmid lacking myeloid zinc finger 1 (MZF1) binding sequence had significantly decreased luciferase activity (P<0.05). Furthermore, the FS protein expression was significantly increased in PK15 cells while the MZF1 was overexpressed, suggesting that the short sequence “TCCCCACC” (the recognition site of transcription factor MZF1) was the most important for FS transcription activation in the porcine.

Association analyses of DNA polymorphisms in immune-related candidate genes GBP1, GBP2, CD163, and CD169 with porcine growth and meat quality traits

Genetic polymorphisms within immunity-related candidate genes in pigs have been identified to control variations in immune functions and/or disease resistance. It has become necessary to evaluate the effects of other genetic markers of economically important traits prior to introducing them into marker-assisted selection programs. In this study, polymorphisms of porcine genes coding Interferon-induced Gunylate binding protein 1 (GBP1), GBP2, CD163, and CD169 were investigated for their association with growth and meat quality traits in a Korean native pig breed -Yorkshire inter-crossed F2 pig population (KY-F2). KY-F2 animals (n=346) have been successfully used for linkage mapping to identify quantitative loci that control meat quality, growth, and immunity traits. In our results, polymorphisms in genes GBP1 and GBP2 showed association with pig growth rate as well as meat quality traits such as crude fat, drip loss, and meat color (yellowness) in the KY-F2 population. The polymorphism in gene CD163 only showed association with crude fat, as a meat quality trait. CD169 gene was associated with pork tenderness. In conclusion, four immune-related genetic markers were validated for their association with growth and meat quality traits to gauge their potential use in a swine selection program. The results warrant further studies in other commercial pig populations.

Effect of skeletal muscle fibers on porcine meat quality at different stages of growth

In this study, we examined changes in meat quality and content of muscle types during porcine growth. The influence of the longissimus dorsi muscle fiber composition on meat quality and the correlation between 2 fiber-typing methods (histochemistry and real-time quantitative polymerase chain reaction) were examined. Type IIx and type IIb fibers accounted for most of the total number of fibers; the proportion of these fibers increased during porcine growth (75.42, 80.09, and 79.88%, respectively, at 3 different stages of growth). There was a strong positive correlation between the 2 fiber-typing methods; the correlation coefficients of type I, IIa, and IIx+IIb fiber contents were 0.65, 0.88, and 0.92, respectively. The a* value of meat color was significantly lower at 98 days and negatively correlated with white fiber content (r = -0.69, P < 0.01). Water-holding capacity decreased during porcine growth. The drip loss parameter was positively correlated with type IIx+IIb fiber content (r = 0.55, P < 0.05). Decreased pH was strongly positively correlated with type IIx+IIb fiber content (r = 0.61, P < 0.01) and negatively correlated with type IIa fiber content (r = -0.44, P < 0.05). Therefore, we found that the composition of muscle fibers influenced the establishment of meat quality and its alteration during the early postmortem period.

Polymorphisms in porcine TRH and TRHR gene and associations with growth and fatness traits

Thyrotropin-releasing hormone (TRH), produced in the hypothalamus and stimulating secretion of thyrotropin from the pituitary, is central in regulating the hypothalamic–pituitary–thyroid (HPT) axis. The number of TRH receptors (TRHR) expressed on the surface of target cell is a crucial factor influencing the magnitude of TRH response. The objective of this study was to identify the DNA polymorphisms existing in pig TRH and TRHR gene and to evaluate their associations with porcine growth, carcass and meat quality traits. Six polymorphisms in TRH and seven in TRHR gene in total were identified by re-sequencing. Among the polymorphisms, TRH ss325994920 was a missense mutation at TRH precursor residue 173 causing Val>Met (GTG>ATG) and would generate mutant pFE22/pSE14 peptides which are non-TRH peptides derived from the TRH precursor and have roles in regulating hormone ACTH and PRL. Allele frequencies of the polymorphisms were evaluated in four Western and two Chinese pig breeds (N=228). Opposite alleles prevailed in Western pigs and Chinese pigs at loci ss325994933, ss325994918, ss325994923, ss325994924 and ss325994925, and existence of high linkage disequilibrium blocks in all six breeds were observed. A resource population (N=409) based on crosses of Pietrain and Jinhua pigs were used for the association tests. TRH gene was significantly (P<0.05) associated with intramuscular fat content, BFT at shoulder and ADG from 150 to 180days old, while TRHR gene was significantly associated with head weight and ADG from 90 to 120days in the Pietrain×Jinhua resource population. Thus, it was suggested that TRH and TRHR could be the candidate genes for growth, meat quality and fatness traits. As extended linkage disequilibrium existed in resource populations, the association results obtained in the present study need further verification. Though verifying test in Yorkshire (N=57), Landrace (N=27) and Duroc (N=30) showed no significant results, the sample number might not be big enough to be statistical powerful. Peripheral tissue expression profile of porcine TRH mRNA was also investigated in this study. Co-expression of TRH and its receptor TRHR in the same tissue, especially in fat tissue, indicated the possibility of the existence of local paracrine/autocrine regulations of TRH on respective peripheral tissue.

Molecular Characterization, Expression Patterns, and Polymorphism of a Differentially Expressed Porcine Gene (PYGM) Isolated by Suppression Subtractive Hybridization and Two-Dimensional Gel Electrophoresis Analysis

Suppression subtractive hybridization was performed to detect the differences in gene expression of porcine longissimus dorsi muscles between Large White and Chinese Meishan pigs. An upregulated gene in Large White that shared high homology with human muscle glycogen phosphorylase (PYGM) was identified. The porcine PYGM gene contains an open reading frame encoding 842 amino acid residues with 26 and 283 nucleotides in the 5' and 3' untranslated regions, respectively. Tissue distribution analysis indicated that porcine PYGM mRNAs are highly expressed in all tissues. Expression pattern of PYGM was similar in the two breeds. Both breeds had the highest expression levels when 120 days old (p<0.01), and PYGM was upregulated during skeletal muscle development. A similar expression pattern of PYGM in protein level was also observed by differential proteome analysis of skeletal muscle development using two-dimensional gel electrophoresis and mass spectroscopy. The mRNA abundance of PYGM in Large White was higher than Meishan at all four stages (p<0.05). Moreover, a G/T mutation in exon 8 was identified and association analysis with meat quality traits showed that it was significantly associated with lean meat percentage (p<0.05). Our data may provide further insight into the molecular mechanisms responsible for breed-specific differences in porcine growth and meat quality.

Mapping QTL in the porcine MHC region affecting fatness and growth traits in a Meishan/Large White composite population

A number of studies have mapped QTL regulating porcine fatness and growth traits to the region of the major histocompatibility complex (MHC) on porcine chromosome 7 using various experimental crosses. The QTL results from crosses using the Chinese Meishan (MS) (slow growing and fat) are particularly interesting because the MS alleles have been found to be associated with increased growth rate and reduced backfat depth. We investigated these QTL further in a composite population derived previously over eight generations by intercrossing Meishan and the European Large White breeds. Genotype information from 32 markers in a 15cM target region was used in linkage and association analyses. A two-step variance component analysis identified QTL for three growth-related traits, explaining 19 ∼ 24% of the phenotypic variance with a confidence interval of 4 cM in the target region. SNP association analyses found that ss181128966 and ss181128924 within the QTL interval were strongly associated with the growth traits. Only weak signals for an effect on backfat depth were found in the association and linkage analyses, possibly because of past directional selection in the composite population.

Ghrelin and Leptin Did Not Improve Meiotic Maturation of Porcine Oocytes CulturedIn Vitro

To improve culture system for in vitro maturation (IVM) of porcine oocytes, ghrelin, leptin or growth hormone (GH), at concentration of 0, 0.5, 5, 50 and 500 ng/ml were added to the porcine follicular fluid (pFF)-supplemented medium NCSU23, and their effects on the maturation and cytoskeletal distribution of the oocytes with or without cumulus cells were compared. In the cumulus-denuded oocytes, no significant changes were noted in the maturation rate by different hormone treatments due to a marked decline in the controls. Maturation of the cumulus intact oocytes was moderately interfered by ghrelin (0.5-50 ng/ml, p < 0.01), but not significantly affected by leptin and GH. Distribution density of the cytoplasmic microtubules was decreased significantly by addition of ghrelin (by approximately 30% in 50-500 ng/ml, p < 0.01), whereas no remarkable effect was noted by leptin supplementation. High concentration (500 ng/ml) of ghrelin or leptin decreased significantly the cytoplasmic microfilaments in density (by 43% and 38%, p < 0.01, respectively). GH did not affect cytoskeletal distribution. The results suggest, in the culture system using pFF-supplemented medium that (i) ghrelin may have some inhibitory effect on the organization of microtubules and microfilaments, probably being a factor in lowered maturation rate and (ii) the addition of higher concentration of leptin may decrease microfilaments in density with no effect on meiotic maturation of the porcine oocytes.

Association of Mahogany/Attractin Gene (ATRN) with Porcine Growth and Fat

Associations of restriction fragment length polymorphism in porcine attractin (ATRN) gene with average daily gain (ADG), live weight (LWT) and back fat thickness (BFT) were studied with one exotic composition population (Xianghuang) and three local breeds (Ningxing, Shaziling and Daweizi) in China. Two types of alleles were detected; one with the band size of 700 bp (A) and the other with the band size of 450 and 250 bp (B), and their genotypes were classified as AA, AB and BB. Polymorphisms were observed in the exotic breed, but not in the three local breeds. Individuals with the genotype BB showed larger ADG and LWT than those with AA in Xianghuang population (p<0.05). The breed specific effects of the porcine attractin gene in this study implied that identification of the polymorphisms and determination of genetic effects on phenotypes should be performed in the target populations. (Asian-Aust. J. Anim. Sci. 2005. Vol 18, No. 10 : 1383-1386)

The effect of some antiproliferative agents on porcine coronary artery function and growth of VSMCs in vitro

The effect of some antiproliferative agents on porcine coronary artery function and growth of VSMCs in vitro

Combined analyses of data from quantitative trait loci mapping studies. Chromosome 4 effects on porcine growth and fatness.

For many species several similar QTL mapping populations have been produced and analyzed independently. Joint analysis of such data could be used to increase power to detect QTL and evaluate population differences. In this study, data were collated on almost 3000 pigs from seven different F(2) crosses between Western commercial breeds and either the European wild boar or the Chinese Meishan breed. Genotypes were available for 31 markers on chromosome 4 (on average 8.3 markers per population). Data from three traits common to all populations (birth weight, mean backfat depth at slaughter or end of test, and growth rate from birth to slaughter or end of test) were analyzed for individual populations and jointly. A QTL influencing birth weight was detected in one individual population and in the combined data, with no significant interaction of the QTL effect with population. A QTL affecting backfat that had a significantly greater effect in wild boar than in Meishan crosses was detected. Some evidence for a QTL affecting growth rate was detected in all populations, with no significant differences between populations. This study is the largest F(2) QTL analysis achieved in a livestock species and demonstrates the potential of joint analysis.

Molecular cloning of porcine α1-microglobulin/HI-30 reveals developmental and tissue-specific expression of two variant messenger ribonucleic acids

A 1008 basepair (bp) cDNA clone encoding 335 amino acids followed by an inframe TGA translation termination codon and a 295-nucleotide 3′ untranslated (UT) region has been isolated from a pig liver cDNA library. Based on the deduced amino acid and nucleotide sequence homology to a human cDNA (Kaumeyer, J.F., Polazzi, J.O. and Kotick, M.P. (1986) Nucleic Acids Res. 14, 7839–7850), the 5′ amino terminus was found to code for α 1-microglobulin ( α 1-M), a 183 amino acid protein belonging to the lipocalin protein superfamily (Pervaiz, S. and Brew, K. (1985) Science 228, 335–337). The 3′ half encoded HI-30 which constitutes the Kunitz-type proteinase inhibitory (L-chain) domain of porcine inter-alpha-trypsin inhibitor (IαTI). In Northern blot hybridization, this cDNA identified two equally abundant mRNA species of approx. 1.3 kb and 1.6 kb in length. However, a 125 bp cDNA probe derived from the 3′ UT region of the cDNA hybridized only to the 1.6 kb mRNA. The differences observed in the 3′ UT region of these mRNAs suggest the utilization of alternative polyadenylation signals or presence of unprocessed nuclear RNA. Densitometric scanning of Northern blots indicated that α 1-M/HI-30 mRNA levels were higher (5–8-fold) in fetal and neonatal liver compared to that of primiparous pigs. In contrast, the RNA levels did not change significantly during pregnancy. Dot blot analysis of RNA indicated liver to be the major site of α 1-M/HI-30 mRNA expression with lower levels observed in the stomach. The results suggest that modulation of α 1-M/HI-30 gene expression could play a role during porcine growth. Increased IαTl L-chain mRNA levels may be particularly important in fetal and neonatal development when regulation of the inflammatory response and protection of macromolecules from proteolytic degradation is vital to survival and sustained growth.

Heteroclitic behaviour of some monoclonal antibodies against bovine growth hormone

Five monoclonal antibodies (MAbs) prepared against bovine growth hormone (bGH) were found to be directed against an immunodominant antigenic domain in bGH. MAbs D8 and H3 reacted equally well with bGH and ovine growth hormone and to a lesser extent with equine (eGH) or porcine (pGH) growth hormones while MAbs H1, H2 and C12 behaved as heteroclitic antibodies, i.e. they bound better a cross-reacting antigen (eGH on pGH) than the immunogen. The reactivity of bGH with the heteroclitic MAbs increased proportionally to the time that the native protein was kept frozen. Deamidation of bGH by treatment with alkali also increased its reactivity. Circular dichroism measurements indicated the occurrence of changes in the conformation of the bGH molecule by these treatments which presumably uncover normally buried or non-accesible epitopes. bGH shares epitopes with eGH and pGH which are immunologically expressed by bGH only when its native conformation is modified.

Relative Potencies of Human, Rat, Bovine/Caprine, Porcine and Ovine Hypothalamic Growth Hormone-Releasing Factors to Release Growth Hormone by the Rat Anterior Pituitary in vitro

Synthetic rat, human, bovine/caprine, porcine and ovine growth hormone-releasing factor (GRF) were tested for their capacity to release growth hormone (GH) by the rat anterior pituitary in vitro. All peptides elicited parallel and dose-dependent increases in GH release and produced similar maximal GH secretion. Rat GRF was 3-6 times more potent in stimulating the release of GH than all other GRFs, while human, bovine/caprine, porcine and ovine GRF had potencies that were not statistically different. The increased potency of both rat GRF(1-27)NH2 and rat GRF(1-23)NH2 when compared to human GRF(1-27)NH2 and human GRF(1-23)NH2, respectively, suggests that the increased potency of this molecule resides in structural differences in the amino terminal of native GRF. The results demonstrate increased sensitivity of rat pituitary cells for their homologous releasing factor.

A Hybrid Noncovalent Complex of Fragments of Porcine and Human Growth Hormones with Diabetogenic Activity*

Although porcine GH (pGH) and human GH (hGH) are structurally related, their structures differ to the extent that, unlike hGH, pGH is not active in primates and does not exhibit lactogenic activity. Therefore, it was of interest to determine whether hybrid noncovalent complexes could be formed by complementation of large fragments of pGH with fragments of hGH and to study the immunological and biological properties of such hybrids. For this purpose, pGH was digested with bovine thrombin conjugated to Sepharose. After reduction and S-carbamidomethylation of the digested hormone, two large fragments consisting of residues 1-133 (p1-133T) and 134-191 (p134-191T) were isolated by gel filtration. The human GH fragments used in this work were S-carbamidomethylated peptides 1-134 (h1-134T) and 135-191 (h135-191T) isolated from thrombin-digested hGH. Noncovalent complementation of the peptide fragments was attempted by dissolving equimolar amounts of the materials in 0.5% (wt/vol) ammonium bicarbonate solution containing 6 M guanidine-HCl and then gradually removing the guanidine-HCl by dialysis. Using this method, a hybrid noncovalent complex of peptides p1-133T and h135-191T was produced in 50% yield by weight of starting material. Attempts to produce the reciprocal complex between h1-134T and p134-191T, as well as attempts to recombine the contiguous pGH fragments, p1-133T, and p134-191T, were unsuccessful. The hybrid complex (p1-133T + h135-191T) had 40% the cross-reactivity of native pGH in a rat GH RIA, but it was inert in an hGH RIA. It had very low growth-promoting activity in the 9-day weight gain test in hypophysectomized rats and no detectable in vitro ability to stimulate [14C]phenylalanine incorporation into the protein of the diaphragm of the hypophysectomized rat. When tested for insulin-like activity (stimulation of glucose oxidation in vitro in adipose tissue of hypophysectomized rats), the hybrid complex had less than 1% the activity of native pGH. In contrast, the hybrid complex had approximately 10-20% the diabetogenic activity of native pGH in the ob/ob mouse. The fact that the hybrid complex has substantial diabetogenic activity, although lacking significant anabolic or insulin-like activities, suggests that the structural requirements differ for the expression of these various activities of GH.

Ovine growth hormone sequence of the C-terminal 68 amino acids

Summary The amino acid sequence of the C-terminal 68 residues of ovine growth hormone is reported. The degree of homology between this region of ovine growth hormone and of bovine, porcine and human growth hormone and ovine prolactin is 99%, 90%, 62% and 22% respectively. The total residue number of ovine growth hormone is approximately 188, corresponding to a molecular weight of 21,300.

EFFECTS OF GROWTH HORMONE ON LEUCINE METABOLISM IN ADIPOSE TISSUE IN VITRO.

Paired samples of rat epididymal adipose tissue were incubated in bicarbonate buffer in the presence of glucose and dl-leucine-2-C14. Bovine growth hormone added to the incubation medium at a concentration of 1 μg/ml doubled the conversion of leucine-C14 to fatty acids in adipose tissue from hypophysectomized rats. Porcine, ovine and human growth hormone preparations had similar effects. However, no such stimulation of leucine metabolism was seen when adipose tissue from normal rats was used. Corticotropin, thyrotropin and oxytocin had no effect on leucine metabolism, but ovine prolactin in high doses (25–100 μg) was found to stimulate the conversion of leucine to fatty acids. Growth hormone also increased the uptake of leucine from the incubation medium but did not influence the oxidation of leucine to CO2.

IN VITRO AND IN VIVO EFFECT OF GROWTH HORMONE ON ALDOSTERONE SECRETION

Porcine, monkey, and human growth hormone have no effect on the in vitro secretion of aldosterone by the rat adrenal gland. When monkey growth hormone is injected into hypophysectomized rats, the adrenals of these animals secrete, under in vitro conditions, increased amounts of aldosterone with no change in the secretion rate of corticosterone. The plasma of these rats contains a substance which appears to stimulate the secretion of aldosterone in the adrenals of normal rats.

IN VITRO AND IN VIVO EFFECT OF GROWTH HORMONE ON ALDOSTERONE SECRETION

Porcine, monkey, and human growth hormone have no effect on the in vitro secretion of aldosterone by the rat adrenal gland. When monkey growth hormone is injected into hypophysectomized rats, the adrenals of these animals secrete, under in vitro conditions, increased amounts of aldosterone with no change in the secretion rate of corticosterone. The plasma of these rats contains a substance which appears to stimulate the secretion of aldosterone in the adrenals of normal rats.