Isolation and molecular characterization of the porcine transforming growth factor beta type I receptor (TGFBR1) gene

Chen, Kefei; Rund, Laurie A.; Beever, Jonathan E.; Schook, Lawrence B.

doi:10.1016/j.gene.2006.07.009

Cited by 18 publications

(14 citation statements)

References 36 publications

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“…SMARCAD1 is required for global deacetylation of histones, methylation of lysines, establishment of heterochromatin and chromosome segregation [40]. TGFBR1 has a ubiquitous expression pattern and is expressed in testes [41] and testicular expression of TGFBR1 is decreased in male lambs that were prenatally exposed to testosterone propionate compared to control male lambs [42]. YOD1 is a highly conserved deubiquitinating enzyme and maintains protein homeostasis in the endoplasmic reticulum.…”

Section: Resultsmentioning

confidence: 99%

Dysregulated microRNA Clusters in Response to Retinoic Acid and CYP26B1 Inhibitor Induced Testicular Function in Dogs

Kasimanickam

Dernell

2014

PLoS ONE

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Spermatogenesis is a multistep synchronized process. Diploid spermatogonia differentiate into haploid spermatozoa following mitosis, meiosis and spermiogenesis. Division and differentiation of male germ cells is achieved through the sequential expression of several genes. Numerous mRNAs in the differentiating germ cells undergo post-transcriptional and translational regulation. MiRNAs are powerful negative regulators of mRNA transcription, stability, and translation and recognize their mRNA targets through base-pairing. Retinoic acid (RA) signaling is essential for spermatogenesis and testicular function. Testicular RA level is critical for RA signal transduction. This study investigated the miRNAs modulation in an RA- induced testicular environment following the administration of all-trans RA (2 µM) and CYP26B1- inhibitor (1 µM) compared to control. Eighty four canine mature miRNAs were analyzed and their expression signatures were distinguished using real-time PCR based array technology. Of the miRNAs analyzed, miRNA families such as miR-200 (cfa-miR-200a, cfa-miR-200b and cfa-miR-200c), Mirlet-7 (cfa-let-7a, cfa-let-7b, cfa-let-7c, cfa-let-7g and cfa-let-7f), miR-125 (cfa-miR-125a and cfa-miR-125b), miR-146 (cfa-miR-146a and cfa-miR-146b), miR-34 (cfa-miR-34a, cfa-miR-34b and cfa-miR-34c), miR-23 (cfa-miR-23a and cfa-miR-23b), cfa-miR-184, cfa-miR-214 and cfa-miR-141 were significantly up-regulated with testicular RA intervention via administration of CYP26B1 inhibitor and all-trans-RA and species of miRNA such as cfa-miR-19a, cfa-miR-29b, cfa-miR-29c, cfa-miR-101 and cfa-miR-137 were significantly down-regulated. This study explored information regarding chromosome distribution, human orthologous sequences and the interaction of target genes of miRNA families significantly distinguished in this study using prediction algorithms. This study importantly identified dysregulated miRNA species resulting from RA-induced spermatogenesis. The present contribution serves as a useful resource for further elucidation of the regulatory role of individual miRNA in RA synchronized canine spermatogenesis.

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Section: Resultsmentioning

confidence: 99%

Dysregulated microRNA Clusters in Response to Retinoic Acid and CYP26B1 Inhibitor Induced Testicular Function in Dogs

Kasimanickam

Dernell

2014

PLoS ONE

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“…Total mRNA was treated with RNase-free DNAse (Qiagen, Valencia, CA) to eliminate genomic contamination. Reverse transcription was then performed using the OmniscriptKit (Qiagen, Valencia, CA) (20). The primers used to quantify the bhmt transcrips (Table 1) were designed using Primer Express software (Applied Biosystems, Foster city, CA).…”

Section: Methodsmentioning

confidence: 99%

“…The negative controls (minus template or reverse transcriptase) were run in triplicate. Data was normalized using 18S ribosomal RNA as an internal control and to the amount of genomic DNA/tissue/sample (20). The parameters used for qPCR were 50°C for 2 min followed by 95°C for 10 min, 40 cycles with 95°C for 15s, 60°C for 1 min, final cycle of 95°C for 15s, 60°C for 15s and 95°C for 15s.…”

Section: Methodsmentioning

confidence: 99%

Splicing variants of the porcine betaine–homocysteine S-methyltransferase gene: Implications for mammalian metabolism

Ganu

Garrow

Koutmos

et al. 2013

Gene

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Betaine-homocysteine S-methyltransferase (BHMT) activity is only detected in the liver of rodents, but in both the liver and kidney cortex of humans and pigs; therefore, the pig was chosen as a model to define the spatial and temporal expression of BHMT during development. During fetal development, a total of ten splice variants of bhmt were expressed at varying levels across a wide range of porcine tissues. Two variants contained an identical ORF that encoded a C-terminal truncated form of BHMT (tBHMT). The bhmt transcripts were expressed at significant levels in the liver and kidney from day 45 of gestation (G45) onward. The transcripts encoding tBHMT represented 5–13% of the total bhmt transcripts in G30 fetus, G45 liver, and adult liver and kidney cortex. The dominant structural feature of wild type BHMT is an (βα)8 barrel, however, a modeled structure of tBHMT suggests this protein would assume a horseshoe fold and lack methyltransferase activity. Low BHMT activity was detected in the G30 fetus, and slightly increased levels of activity were observed in the liver from G45 and G90 fetuses. The bhmt promoter contained three key CpG sites, and methylation of these sites was significantly higher in adult lung compared to adult liver. The data reported herein suggest that genomic DNA methylation and variation of the 5' and 3' UTRs of bhmt transcripts are key regulators for the level of BHMT transcription and translation.

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“…Each tissue was collected from the same three pigs and for each sample three replicates were performed. RNA was extracted, followed by reverse transcription with the OmniscriptKit (Qiagen, Valencia, CA) using the protocol of Chen et al (2006). Primers (Table 1) were designed using Primer Express software (Applied Biosystems, Foster city, CA).…”

Section: Methodsmentioning

confidence: 99%

“…The total number of BHMT and BHMT-2 transcripts in a given tissue was determined using the following primer sets; qBHMT-f and qBHMT-r, and qB2-f and qB2-r, respectively. Total 18s rRNA was used as an internal control (Chen et al 2006) and each reaction contained 100ng of cDNA. Negative controls (minus template or reverse transcriptase) were also run in triplicate.…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization and analysis of the porcine betaine homocysteine methyltransferase and betaine homocysteine methyltransferase-2 genes

Ganu

Garrow

Sodhi

et al. 2011

Gene

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Betaine homocysteine methyltransferase (BHMT) and BHMT-2 enzymes methylate homocysteine to form methionine using betaine and S-methylmethionine, respectively. These activities are observed only in the liver of adult rodents, but in adult humans and pigs these activities are detected in both the liver and kidney, indicating the pig is a more appropriate model for studying the biochemical and physiological roles of these enzymes in human biology. Porcine BHMT and BHMT-2 cDNAs were cloned and sequenced, and their 5' and 3' UTR were amplified using RLM-RACE. The BHMT transcript had significantly longer 5' and 3' UTRs than BHMT-2. The pig BHMT and BHMT-2 genes span approximately 26kb and 16kb, respectively, and both genes have 8 exons. The deduced amino acid sequences of BHMT and BHMT-2 contain 407 and 363 amino acids, respectively, and shared 78% amino acid identity. No promoter element (TATA or CAAT box) was observed for either BHMT or BHMT-2, although a CpG island surrounding the promoter and transcriptional start site were observed in both genes implying that methylation could regulate their expression. Using qPCR, it was determined that BHMT and BHMT-2 transcripts are very abundant in liver and kidney cortex, whereas the expression is significantly less in other tissues. These findings confirm that the expression pattern of BHMT and BHMT-2 genes in pigs is similar to humans, supporting the use of the pig as an animal model to study the genetics and regulation of BHMT and BHMT-2 expression.

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Isolation and molecular characterization of the porcine transforming growth factor beta type I receptor (TGFBR1) gene

Cited by 18 publications

References 36 publications

Dysregulated microRNA Clusters in Response to Retinoic Acid and CYP26B1 Inhibitor Induced Testicular Function in Dogs

Dysregulated microRNA Clusters in Response to Retinoic Acid and CYP26B1 Inhibitor Induced Testicular Function in Dogs

Splicing variants of the porcine betaine–homocysteine S-methyltransferase gene: Implications for mammalian metabolism

Molecular characterization and analysis of the porcine betaine homocysteine methyltransferase and betaine homocysteine methyltransferase-2 genes

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