Developmental Changes of Growth Hormone Mrna Abundance in Pituitary Tissue of Jinhua and Landrace Gilts

2011 ◽  
Vol 343-344 ◽  
pp. 412-416
Author(s):  
Zhi Guo Miao ◽  
Guo Wang Li ◽  
Shi Zhu Wang ◽  
Xin Yao Chang ◽  
Hong Bing Xie ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Mrna Expression ◽  
Age Groups ◽  
Rt Pcr ◽  
Developmental Patterns ◽  
Expression Levels ◽  
Pituitary Tissue ◽  
Breed Differences ◽  
Gh Gene ◽  
Mrna Expression Levels

In this pepar we investigated the developmental patterns of expression of growth hormone (GH) gene in pituitary tissue in pigs of different breeds and their effects on the carcass fat contents. 3 Jinhua gilts and 3 Landrace gilts were sampled at 35, 80 and 125 days of age, respectively. Carcass fat contents were determined. Pituitary tissue was sampled and total RAN was extracted to determine GH mRNA expression levels by semi-quantitative RT-PCR. The results showed that the contents of carcass fat increased with growth and showed significant differences (P﹤0.05) between different age groups in the two breeds. Furthermore, carcass fat contents in Jinhua gilts were higher than that in Landrace gilts during growth (P﹤0.05). GH mRNA expression levels decreased with age and displayed breed differences. Jinhua gilts showed lower abundance of GH mRNA compared with Landrace gilts at 35, 80 and 125 days of age (P﹤0.05). In addition, GH mRNA expression level was negatively related to carcass fat content in Jinhua and Landrace gilts (r = -0.790 (P = 0.01), r = -0.755 (P = 0.02), respectively).

Comparison of mRNA Expression Levels Determined with TAQ?Man and Competitive Template RT?PCR

ChemInform ◽  
2005 ◽  
Vol 36 (12) ◽  
Author(s):  
R. Mauritz ◽  
W. Schwabe ◽  
P. Haeusler ◽  
P. Noordhuis ◽  
K. Smid ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Rt Pcr ◽  
Expression Levels ◽  
Competitive Template ◽  
Mrna Expression Levels

Comparison of mRNA Expression Levels Determined with TAQ–Man and Competitive Template RT–PCR

2004 ◽  
Vol 23 (8-9) ◽  
pp. 1471-1474 ◽  
Author(s):  
R. Mauritz ◽  
W. Schwabe ◽  
P. Haeusler ◽  
P. Noordhuis ◽  
K. Smid ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Rt Pcr ◽  
Expression Levels ◽  
Competitive Template ◽  
Mrna Expression Levels

scholarly journals MON-603 GPR142 Expression Levels Were Correlated with Plasma Ghrelin Levels and Heights in Morbidly Obese Patients

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Yoko Ueda ◽  
Hiroshi Iwakura ◽  
Asako Doi ◽  
Norihiko Matsutani ◽  
Shuhei Morita ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Mrna Expression ◽  
Morbidly Obese ◽  
Obese Patients ◽  
Expression Levels ◽  
Desacyl Ghrelin ◽  
Morbid Obese ◽  
Ghrelin Secretion ◽  
Mrna Expression Levels

Abstract Recently, aromatic amino acids, especially tryptophan were discovered to be the strongest ligands for GPR142, which was previously known as an orphan GPCR. GPR142 is expressed in the digestive tract and pancreas in mice and human. Previously we found that GPR142 is highly expressed in the ghrelin-producing cell line, MGN3-1 cells, and that tryptophan strongly stimulated ghrelin secretion in vitro. In this study, we measured the mRNA expression levels of GPR142 in the gastric samples of 6 morbid obese patients undergone laparoscopic sleeve gastrectomy and compared its level with their clinical parameters. GPR142 expression levels were negatively correlated with plasma desacyl ghrelin levels (p=0.011) and positively correlated with heights (p=0.08). The current results that GPR142 expression levels were correlated with plasma desacyl ghrelin levels may confirm the link between GPR142 signal and regulation of ghrelin secretion demonstrated in our in vitro study. Regarding to the correlation with heights, there are some reports that plasma ghrelin levels were inversely correlated with heights in children[1-3], although, as far as we know, there are no reports demonstrating the relationship between plasma ghrelin levels and heights in adults. Considering that ghrelin strongly stimulates growth hormone secretion, GPR142 signaling may have influence on height through regulating ghrelin-growth hormone axis. Conclusion GPR142 mRNA expression levels were negatively and positively correlated with plasma desacyl ghrelin levels and heights in morbid obese adults undergone bariatric surgery. Current results may help understanding the pathophysiological role of GPR142 in the regulation of ghrelin secretion and heights. References 1. MO Camurdan, et al. Endocrine Journal 2006, 53 (4), 479–484 2. HS Park, et al. Metabolism Clinical and Experimental 2005, 54, 925–929 3. Joy C. Bunt,et al. J Clin Endocrinol Metab 2003, 88(8): 3756-3761

scholarly journals β-Sitosterol 3-O-D-glucoside increases ceramide levels in the stratum corneum via the up-regulated expression of ceramide synthase-3 and glucosylceramide synthase in a reconstructed human epidermal keratinization model

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0248150
Author(s):  
Shogo Takeda ◽  
Shuko Terazawa ◽  
Hiroshi Shimoda ◽  
Genji Imokawa
Keyword(s):  
Stratum Corneum ◽  
Mrna Expression ◽  
Acid Sphingomyelinase ◽  
Pcr Analysis ◽  
Ceramide Synthase ◽  
Rt Pcr ◽  
Glucosylceramide Synthase ◽  
Expression Levels ◽  
Regulated Expression ◽  
Mrna Expression Levels

β-Sitosterol 3-O-d-glucoside (BSG) is known to act as an agonist by binding to estrogen receptors, and estrogen has been reported to enhance the activity of β-glucocerebrosidase, an epidermal ceramide metabolizing enzyme. In this study, we determined whether BSG up-regulates ceramide levels in the stratum corneum (SC) of a reconstructed human epidermal keratinization (RHEK) model. Treatment with BSG significantly increased the total ceramide content by 1.2-fold compared to that in the control in the SC of the RHEK model, accompanied by a significant increase of the ceramide species, Cer[EOS] by 2.1-fold compared to that in the control. RT-PCR analysis demonstrated that BSG significantly up-regulated the mRNA expression levels of serine palmitoyltransferase (SPT)2, ceramide synthase (CerS)3, glucosylceramide synthase (GCS) and acid sphingomyelinase by 1.41–1.89, 1.35–1.44, 1.19 and 2.06-fold, respectively, compared to that in the control in the RHEK model. Meanwhile, BSG significantly down-regulated the mRNA expression levels of sphingomyelin synthase (SMS)2 by 0.87–0.89-fold. RT-PCR analysis also demonstrated that BSG significantly up-regulated the mRNA expression levels of CerS3 and GCS by 1.19–1.55 and 1.20-fold, respectively, but not of SPT2 and significantly down-regulated that of SMS2 by 0.74-fold in HaCaT keratinocytes. Western blotting analysis revealed that BSG significantly increased the protein expression levels of CerS3 and GCS by 1.78 and 1.28–1.32-fold, respectively, compared to that in the control in HaCaT cells. These findings indicate that BSG stimulates ceramide synthesis via the up-regulated expression levels of CerS3 and GCS in the glucosylceramide pathway, which results in a significantly increased level of total ceramides in the SC accompanied by significantly increased levels of acylceramide species such as Cer[EOS].

scholarly journals IDENTIFIKASI IKAN CUPANG (Betta imbelis) TRANSGENIK FOUNDER MEMBAWA GEN PENYANDI HORMON PERTUMBUHAN; Identification of Transgenic Founder Betta Fish (Betta imbelis) Carry Growth Hormone Gene.

2017 ◽  
Vol 11 (3) ◽  
pp. 197
Author(s):  
Eni Kusrini ◽  
Alimuddin Alimuddin ◽  
Mohammad Zairin ◽  
Dinar Tri Sulistyowati
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Mrna Expression ◽  
Transgenic Fish ◽  
Pcr Analysis ◽  
Growth Hormone Gene ◽  
Rt Pcr ◽  
Total Rna ◽  
Gh Gene ◽  
Dna Template

Penelitian dilakukan untuk mengidentifikasi keberhasilan introduksi gen penyandi hormon pertumbuhan (Growth Hormone, GH) pada induk F-0 ikan Betta imbellis. Ikan transgenik F-0 dibuat dengan menggunakan metode transfeksi. Identifikasi dilakukan menggunakan metode RT-PCR. RNA total diekstraksi dari embrio pooled sample hasil persilangan induk transgenik dan non-transgenik. Berdasarkan analisis ekspresi gen pada embrio juga menunjukkan adanya aktivitas ekspresi gen GH pada semua perlakuan dibandingkan dengan kontrol (embrio hasil persilangan non-transgenik x non-transgenik). Jumlah individu induk F-0 yang membawa gen GH eksogen berdasarkan analisis PCR dengan DNA template dari sirip ekor adalah sebanyak 16%. Individu positif membawa gen GH eksogen tersebut dibesarkan lebih lanjut untuk memproduksi Betta imbellis transgenik F-1. Kandidat ikan transgenik jantan F-0 dikawinkan dengan ikan non-transgenik betina, sedangkan transgenik F-0 betina dikawinkan dengan non-transgenik jantan. Sebanyak 30-50 butir embrio hasil pemijahan F-0 digabung, kemudian DNA genom diekstrak. Sebagian embrio digunakan untuk ekstraksi RNA total untuk analisis ekspresi mRNA GH eksogen. Hasil analisis PCR menunjukkan bahwa semua sampel embrio dari induk transgenik F-0 dapat terdeteksi gen GH eksogen, sedangkan untuk kontrol (non-transgenik) tidak terdeteksi. Ekspresi mRNA juga terdeteksi pada embrio F-1. Dengan demikian, metode transfeksi embrio Betta imbellis efektif digunakan untuk menghasilkan ikan transgenik, dan sangat berpotensi menghasilkan individu F-1 Betta imbellis dengan pertumbuhan lebih cepat.The study was conducted to identify the successful introduction of the growth hormone gene (Growth Hormone, GH) on the F-0 Betta imbellis broodstock. The F-0 transgenic fish was made through transfection methods. Identification was done using RT-PCR method. Total RNA was extracted from pooled embryos sample. Based on the analysis of gene expression in embryos also showed activity GH gene expression in all treatments compared to the control (non-transgenic x non-transgenic). The number of individuals F-0 which carried exogenous GH gene by PCR analysis of the DNA template of the tail fin was as much as 16%. Positive individuals carried the exogenous GH gene raised further to produce transgenic F-1 B. imbellis. Candidate of transgenic F-0 males fish were mated with non-transgenic female fish, whereas the transgenic F-0 females were mated with non-transgenic males. The 30-50 embryos obtained were combined, then their genomic DNA were extracted. Some of the embryos was used for the extraction of total RNA for analysis of mRNA expression of GH exogenous. The PCR analysis showed that all samples of embryos from the transgenic F-0 broodstock could be detected, whereas for the control (non-transgenic) was not detected. mRNA expression was also detected in embryos of F-1. The average weight of the F-0 broodstocks were 1.55 g and a total length was 12.97 cm. Thus, the transfection methods through betta embryos peaceful effectively generated transgenic fish, and potentially produced fast growth of individuals F-1 Betta imbellis.

Targeted Reduction of Let-7a miRNA Increases Fetal Hemoglobin in Human Adult Erythroblasts

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 451-451 ◽  
Author(s):  
Jaira F. de Vasconcellos ◽  
Colleen Byrnes ◽  
Y. Terry Lee ◽  
Megha Kaushal ◽  
Joshua M. Allwardt ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Mirna Family ◽  
Fetal Hemoglobin ◽  
Rt Pcr ◽  
Globin Mrna ◽  
Expression Levels ◽  
Gamma Globin ◽  
Total Hemoglobin ◽  
Cd34 Cells ◽  
Mrna Expression Levels

Abstract MicroRNAs (miRNAs) are a class of small, noncoding RNAs that bind and regulate target messenger RNAs (mRNAs). The let-7 family consists of twelve genes encoding nine highly conserved miRNAs that are involved in developmental timing events in multicellular organisms. Previous studies showed regulation during the fetal-to-adult transition in the erythroid lineage with significant increases in let-7 miRNAs from adult compared to umbilical cord blood reticulocytes (1). Further studies indicated that reduced expression of let-7 in adult CD34+ cells by “sponge” targeting the miRNA family seed region caused increased fetal hemoglobin (HbF), but the mean level of HbF remained less than 20% of the total hemoglobin (2). Increased expression of LIN28A (a major regulator of all let-7 miRNAs) caused greater increases in HbF (greater than 30% of the total) in cultured erythrocytes from pediatric patients with HbSS genotype (3). However, these studies did not address the potential for targeting an individual let-7 miRNA family member to regulate HbF expression. For this purpose, we initially determined the expression levels of mature let-7 family members in purified cell populations sorted from peripheral blood. The total levels of let-7 miRNAs in peripheral blood cells were as follows: reticulocytes: 1.7E+08 ± 1.0E+08 copies/ng; neutrophils: 2.0E+07 ± 1.1E+07 copies/ng; lymphocytes: 1.1E+07 ± 6.2E+06 copies/ng and monocytes: 3.5E+06 ± 2.7E+06 copies/ng. Among the individual species, let-7a was identified as a predominantly expressed let-7 family member in reticulocytes. As such, we hypothesized that specifically targeting let-7a may be sufficient to regulate HbF levels. To study the effects of let-7a miRNAs upon erythropoiesis and globin expression, a lentiviral construct that incorporated the tough decoy (TuD) design to target let-7a was compared with empty vector controls. Transductions were performed in CD34+ cells from five adult healthy volunteers cultivated ex vivo in erythropoietin-supplemented serum-free media for 21 days. Down-regulation of let-7a was confirmed by Q-RT-PCR at day 14 (control: 1.4E+07 ± 2.4E+06 copies/ng; let-7a-TuD: 1.6E+06 ± 4.6E+05 copies/ng; p=0.0003). Cell proliferation and differentiation were comparable in let-7a-TuD versus control transductions. Expression levels of globin genes were evaluated upon let-7a-TuD by Q-RT-PCR. Let-7a-TuD transductions caused significantly increased gamma-globin mRNA expression levels compared to control transductions (control: 1.2E+06 ± 6.8E+05 copies/ng; let-7a-TuD: 1.1E+07 ± 4.5E+06 copies/ng; p=0.004). HPLC analyses at the end of the culture period demonstrated robust increases in HbF levels after let-7a-TuD transduction (HbF control: 4.7 ± 0.6%; let-7a-TuD: 38.2 ± 3.8%; p=0.00003). In addition, the expression patterns of the erythroid transcription factors BCL11A, KLF1 and SOX6 were investigated. Let-7a-TuD decreased BCL11A mRNA expression levels (control: 1.7E+03 ± 4.5E+02 copies/ng; let-7a-TuD: 4.3E+02 ± 1.8E+02 copies/ng; p=0.003), but major changes in KLF1 or SOX6 were not detected. In summary, we report here that the let-7 miRNA family is differentially expressed in purified cell populations from adult human blood, and that let-7a is a predominantly expressed species in reticulocytes. Further, targeted reduction of let-7a in erythroblasts is sufficient to cause robust increases in gamma-globin mRNA expression and HbF to mean levels around 35-40% of the total hemoglobin produced. Targeting of individual let-7 genes or RNA transcripts may be useful for therapeutic induction of HbF expression in patients with sickle cell disease or other beta-hemoglobinopathies. 1) Noh SJ et al. J Transl Med. 7:98 (2009). 2) Lee YT et al. Blood. 122:1034-41 (2013). 3) Vasconcellos JF et al. Blood. 122: Abstract 313 (2013). Disclosures No relevant conflicts of interest to declare.

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