scholarly journals Elastin mRNA levels during foetal development of sheep nuchal ligament and lung. Hybridization to complementary and cloned DNA

1984 ◽  
Vol 220 (3) ◽  
pp. 653-663 ◽  
Author(s):  
J M Davidson ◽  
S Shibahara ◽  
C Boyd ◽  
M L Mason ◽  
P Tolstoshev ◽  
...  
Keyword(s):  
Cdna Probe ◽  
Northern Analysis ◽  
Northern Hybridization ◽  
Mrna Levels ◽  
Foetal Development ◽  
Total Rna ◽  
Mrna Content ◽  
Near Term ◽  
Nuchal Ligament ◽  
Elastin Gene

Elastin mRNA levels were quantified in sheep nuchal ligament and lung during the latter half of foetal development with elastin-specific cDNA (complementary DNA) probes using both hybridization in solution (saturation analysis) and hybridization on a fixed support (Northern analysis). For the solution-hybridization studies, cDNA prepared from nuchal-ligament mRNA was enriched to 65% for elastin sequences by hybridizing it to its template at a R0t (mol X s X litre-1) value that included only the abundant class of mRNA sequences. Hybridization of this probe to RNA extracted from nuchal ligament between 70 and 138 days after conception demonstrated elastin sequences increased about 10-fold (from 0.047 to 0.438% of total RNA). In contrast, lung elastin mRNA levels increased only 3-fold (from 0.009 to 0.022% of total RNA) during the same period. Over this development period these values correspond to increases in the average number of elastin mRNA molecules from 950 to 20 000 molecules/ligament cell and from 130 to 330 molecules/lung cell. For Northern analysis, elastin mRNA was purified from near-term-sheep nuchal ligament on sucrose density gradients. Analysis of the translation products of this elastin mRNA showed that relative elastin precursor synthesis was at least 80% of total [3H]valine incorporation. The Mr of this elastin mRNA, determined by methylmercury-agarose-gel electrophoresis, was approx. 1.25 × 10(6). Northern hybridization of nuchal ligament and lung RNA to a [32P]cDNA probe, transcribed from this sucrose-gradient-purified elastin mRNA, confirmed the developmental changes in elastin mRNA levels detected by solution-hybridization techniques. The specificity of this method was confirmed by using a cloned elastin gene fragment. These studies demonstrate that elastin mRNA levels in organs such as nuchal ligament and lung increase with foetal development, but that there are significant differences in the average cellular elastin mRNA content of these two organs.

Characterization of growth hormone gene expression in the pituitary and plasma growth hormone concentrations during posthatch development in the chicken

1995 ◽  
Vol 145 (2) ◽  
pp. 343-353 ◽  
Author(s):  
K Reiprich ◽  
E Mühlbauer ◽  
E Decuypere ◽  
R Grossmann
Keyword(s):  
Gene Expression ◽  
Growth Hormone ◽  
Rapid Growth ◽  
Northern Analysis ◽  
Northern Hybridization ◽  
Mrna Levels ◽  
Early Stages ◽  
Plasma Gh ◽  
Observation Period

Abstract In this study both sexes of two strains of chicken with genetically different growth potentials (broiler- and laying-type) were used to investigate growth hormone (GH) gene expression during posthatch development from day 7 (D7) to D56 by using the in situ hybridization technique and Northern analysis. In pituitaries of both strains a high GH mRNA signal was found as early as D7 by in situ hybridization, showing clear differences in the pattern of gene expression between the two strains. By Northern hybridization sex differences were detectable in all age groups of broilers, with higher levels throughout in males. In layers, however, females showed consistently higher levels compared with males until D21. While signal intensities decreased in the broiler strain during the investigation period, the layer-type strain seemed to express GH mRNA more continuously, reaching significantly (P<0·01) higher GH mRNA levels than broilers at D56. Plasma GH concentrations ran parallel to GH mRNA in early stages but showed a peak earlier at D14 and decreased after D35 in both sexes and strains. Determination of growth as weekly weight gains, however, proved that a period of rapid growth (at a higher level in both sexes of the broiler strain) at D7 was followed by a strong decrease from D14 to D21. A plateau of constant growth was reached until the end of the observation period with similar rates in both strains and sexes. Analysis of plasma thyroid hormones tri-iodothyronine/thyroxine (T3/T4) showed an increase in T3 concentrations in both strains and sexes in early stages and a decrease thereafter. No clear strain differences were measured. T4 plasma concentrations increased from D7 to D14 in broilers and D21 in layers when a plateau was reached. From the results we conclude that generally there is a good correlation between GH mRNA and plasma GH concentrations in both strains investigated. Neither parameter, however, is coupled directly with the growth rate. Thus the early rapid growth corresponds to relatively low levels of GH mRNA and plasma GH concentrations, but high T3 levels. Later, decreased growth rates are linked to increasing amounts of GH mRNA as well as increasing plasma GH concentrations in both layers and broilers. Towards the end of the observation period there was a strain divergence visible with increased amounts of GH mRNA in layers but a strong reduction in broilers. Moreover, plasma GH concentrations decreased more slowly in layers than in broilers. Journal of Endocrinology (1995) 145, 343–353

scholarly journals Analysis of von Willebrand factor mRNA from the lung of pigs with severe von Willebrand disease by using a human cDNA probe

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1341-1346 ◽  
Author(s):  
QY Wu ◽  
BR Bahnak ◽  
L Coulombel ◽  
D Kerbiriou-Nabias ◽  
L Drouet ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Cdna Probe ◽  
Von Willebrand Disease ◽  
Northern Analysis ◽  
Cdna Clones ◽  
Total Rna ◽  
Human Cdna ◽  
Actin Mrna ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract To examine the control of porcine von Willebrand factor (vWF) biosynthesis we cloned human vWF complementary DNA (cDNA) and investigated the expression of the vWF gene in lungs from normal pigs and pigs with severe von Willebrand's disease (vWD). Recombinant clones spanning approximately 90% of human vWF cDNA were identified in a lambda gt10 human lung cDNA library by screening with oligonucleotides. One clone spanning nucleotides 960 to 3,240 of human vWF cDNA was used to investigate the steady-state levels of vWF mRNA in lungs from normal pigs and from pigs phenotypically determined to be homozygous for vWD. This clone hybridized with genomic DNA from pig leukocytes when Southern blots were processed under very stringent conditions; therefore, human cDNA clones were considered valid probes to detect porcine mRNA. Northern blot analysis of total RNA from normal pig lung and human umbilical vein endothelial cells identified the vWF mRNA as a molecular species of approximately 9.0 kilobases (kb). A very faint to undetectable band at 9.0 kb in total RNA from lungs of vWD pigs suggested a decreased rate of transcription of the vWF gene. Sucrose density gradient centrifugation of RNA from the vWD pigs confirmed by Northern analysis that the high-molecular weight fractions contained vWF mRNA and at the same size as normal pig mRNA. Dot blot hybridization analysis of vWF and actin mRNA processed under stringent conditions demonstrated that the relative ratio of vWF mRNA to actin mRNA in the vWD pigs varied from 21% to 41% of the ratio observed in normal pigs. Because the amount of vWF mRNA is not correlated to the amount of vWF activity or antigen in plasma of vWD pigs we conclude that posttranscriptional events are also probably involved in abnormal expression of vWF in these animals.

The renal endothelin system in the Prague hypertensive rat, a new model of spontaneous hypertension

1999 ◽  
Vol 97 (1) ◽  
pp. 91-98 ◽  
Author(s):  
Volker VOGEL ◽  
Angela BÄCKER ◽  
Jiri HELLER ◽  
Herbert J. KRAMER
Keyword(s):  
Renal Cortex ◽  
Renal Tissue ◽  
Spontaneous Hypertension ◽  
Mrna Levels ◽  
New Model ◽  
Total Rna ◽  
Peptide Concentration ◽  
Hypertensive Rat ◽  
Mrna Content ◽  
Prague Hypertensive Rat

In a new model of spontaneous hypertension, namely the Prague hypertensive rat (PHR), hypertension is transferred with a kidney transplanted from the PHR to its normotensive counterpart (PNR) by an as yet unknown mechanism. One candidate may be endothelin (ET), since this potent vasoconstrictor affects vascular tone, renal haemodynamics and renal excretory function, and all members of this peptide family are located within the kidney and act in an autocrine/paracrine fashion. In the present study we investigated, in the renal tissue of PHRs and PNRs: (1) preproET-1 and preproET-3 mRNAs as well as ET-1 and ET-3 peptide distribution, (2) endothelin-converting enzyme (ECE)-1 mRNA expression, and (3) ET receptors and their characteristics in membranes of glomeruli and papillae. In addition, plasma ET concentration and urinary ET excretion were determined. Quantitative measurements by competitive reverse transcription-polymerase chain reaction revealed ET-1 mRNA levels in the renal cortex from PHRs and PNRs of 1.09±0.13 and 1.29±0.18 amol/µg of total RNA respectively, and in red medulla of 2.72±0.82 and 3.30±0.68 amol/µg respectively. In contrast, renal papilla from PHRs showed significantly lower levels of preproET-1 mRNA (1.81±0.64 amol/µg of total RNA, compared with 4.25±0.82 amol/µg in PNRs; each n = 5; P< 0.05). The ET-1 peptide concentration in papillary tissue was also significantly lower in PHRs than in PNRs (120.2±30.8 and 491.3±53.4 fmol/mg of protein respectively; n = 5; P< 0.01), whereas it was similar in cortex and medulla from PHRs and PNRs. The preproET-3 mRNA content in renal tissue was much lower than that of preproET-1 mRNA. It was significantly higher in red medulla from PHRs compared with that from PNRs (0.25±0.05 and 0.13±0.02 amol/µg of total RNA respectively; P< 0.05), but was similar in papillae of PHRs and PNRs (0.04±0.02 and 0.05±0.01 amol/µg respectively; n = 5). Cortical preproET-3 mRNA was at the lower limit of detection. Similarly, the ET-3 peptide concentration was slightly but significantly higher in the red medulla of PHRs compared with PNRs (15.4±2.0 and 8.8±0.8 fmol/mg of protein respectively; n = 5; P< 0.05), whereas no differences in ET-3 peptide concentration were found in papillae from PHRs and PNRs. ECE-1 mRNA levels were similar in the renal cortex, red medulla and papillae from PHRs and PNRs, ranging between 0.34±0.03 and 0.56±0.12 amol/µg of total RNA. Of the total ET receptors in glomerular membranes, 39% were ETA receptors, whereas papillary membranes contained exclusively ETB receptors. PHRs and PNRs showed similar Bmax and Kd values for ET-1 in renal glomerular membranes (Bmax, 6.5±1.3 and 4.9±1.2 pmol/mg of protein respectively; Kd, 0.69±0.10 and 0.56±0.10 nM respectively) and papillary membranes (Bmax, 9.7±1.1 and 11.3±1.6 pmol/mg of protein respectively; Kd, 0.30±0.04 and 0.42±0.07 nM respectively). Plasma ET-1/2 concentrations (10.4±1.3 and 12.2±1.2 fmol/ml in PHRs and PNRs respectively) and urinary ET-1 excretion (3.1±0.3 and 3.0±0.2 pmol/24 h in PHRs and PNRs respectively) were similar in hypertensive and normotensive rats. In summary, although tissue levels of preproET-3 mRNA were very low in the kidney, significantly greater amounts of preproET-3 mRNA and ET-3 peptide were found in medullary tissue from PHRs compared with PNRs, a finding that awaits further investigation. In contrast, the preproET-1 mRNA content and ET-1 peptide concentration were significantly lower in papillary tissue from PHRs compared with PNRs. Decreased synthesis of ET-1, which normally antagonizes the action of [Arg8]vasopressin, may allow increased water (and sodium) reabsorption at the level of the inner medullary collecting duct. This intrinsic defect of the kidney in the PHR may contribute to hypertension in this model, and may transmit high blood pressure on transplantation of the ‘hypertensive’ kidney into a normotensive rat.

scholarly journals Analysis of von Willebrand factor mRNA from the lung of pigs with severe von Willebrand disease by using a human cDNA probe

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1341-1346
Author(s):  
QY Wu ◽  
BR Bahnak ◽  
L Coulombel ◽  
D Kerbiriou-Nabias ◽  
L Drouet ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Cdna Probe ◽  
Von Willebrand Disease ◽  
Northern Analysis ◽  
Cdna Clones ◽  
Total Rna ◽  
Human Cdna ◽  
Actin Mrna ◽  
Von Willebrand ◽  
Willebrand Factor

To examine the control of porcine von Willebrand factor (vWF) biosynthesis we cloned human vWF complementary DNA (cDNA) and investigated the expression of the vWF gene in lungs from normal pigs and pigs with severe von Willebrand's disease (vWD). Recombinant clones spanning approximately 90% of human vWF cDNA were identified in a lambda gt10 human lung cDNA library by screening with oligonucleotides. One clone spanning nucleotides 960 to 3,240 of human vWF cDNA was used to investigate the steady-state levels of vWF mRNA in lungs from normal pigs and from pigs phenotypically determined to be homozygous for vWD. This clone hybridized with genomic DNA from pig leukocytes when Southern blots were processed under very stringent conditions; therefore, human cDNA clones were considered valid probes to detect porcine mRNA. Northern blot analysis of total RNA from normal pig lung and human umbilical vein endothelial cells identified the vWF mRNA as a molecular species of approximately 9.0 kilobases (kb). A very faint to undetectable band at 9.0 kb in total RNA from lungs of vWD pigs suggested a decreased rate of transcription of the vWF gene. Sucrose density gradient centrifugation of RNA from the vWD pigs confirmed by Northern analysis that the high-molecular weight fractions contained vWF mRNA and at the same size as normal pig mRNA. Dot blot hybridization analysis of vWF and actin mRNA processed under stringent conditions demonstrated that the relative ratio of vWF mRNA to actin mRNA in the vWD pigs varied from 21% to 41% of the ratio observed in normal pigs. Because the amount of vWF mRNA is not correlated to the amount of vWF activity or antigen in plasma of vWD pigs we conclude that posttranscriptional events are also probably involved in abnormal expression of vWF in these animals.

VON WILLEBRAND FACTOR mRNA IS SEVERELY REDUCED IN PIGS WITH HOMOZYGOUS VON WILLEBRAND DISEASE

1987 ◽  
Author(s):  
Q Y Wu ◽  
B R Bahnak ◽  
L Coulombel ◽  
J P Caen ◽  
G Pietu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Von Willebrand Factor ◽  
Limit Of Detection ◽  
Cdna Probe ◽  
Von Willebrand Disease ◽  
Northern Hybridization ◽  
Transcriptional Level ◽  
Total Rna ◽  
Von Willebrand ◽  
Willebrand Factor

Porcine von Willebrand disease (vWD), an autosomal recessive disorder, is similar to some of the severe forms of vWD in humans and is characterized by a prolonged bleeding time and very low or undetectable amounts of von Willebrand factor (vWF) antigen and activity in plasma, platelets and endothelial cells. The molecular events that control the lack of expression of vWF in the vWD pigs is not known and could be at the transcriptional or post-transcriptional level. Lungs from normal and two homozygous vWD pigs were extracted immediately after harvesting of the animals and placed on dry ice. Tissues were homogenized in 6 M guanidinium thiocyanate and RNA isolated by centrifugation through cesium chloride. Total RNA was analyzed by Northern hybridization including dénaturation in glyoxal, electrophoresis in 1.0 % agarose-2.2 M formaldehyde gels and transfer onto nitrocellulose. Messenger RNA was detected with a nick-translated human vWF cDNA probe or a human actin control probe. The vWF probe, cloned from a human lung library, was 2,280 bp in length and spanned nucleotides 960 to 3,240 of the human cDNA. These human probes were considered valid to detect levels of porcine vWF and actin mRNA because they hybridized with restriction enzyme digested genomic DNA from normal and vWD pig leucocytes under conditions of high stringency. The size of the vWF mRNA in the normal pigs after Northern hybridization was approximately 9.0 kb, similar to that of human vWF mRNA, and was easily detectable at the lowest concentration of RNA blotted (5 ug). In contrast, vWF mRNA from vWD pigs was at the lower limit of detection even at 10 ug of total RNA blotted. Nevertheless, although at extremely low levels, vWF mRNA from vWD pigs appeared to be the same size as the normal mRNA. These results agree with observations on the relationship of vWF secreted from 24 hr. cultures of endothelial cells from the pulmonary artery of normal and vWD pigs where the vWF levels were 0.90 and 0.06 U/108 cells, respectively. Therefore, it appears that the very low expression of vWF in the vWD pigs is due to a lack of transcription of the vWF gene. At this time, however, turnover of unstable transcripts in the vWD pigs can not be ruled out.

scholarly journals Regulation of mdr2 P-glycoprotein expression by bile salts

1997 ◽  
Vol 321 (2) ◽  
pp. 389-395 ◽  
Author(s):  
Charles M. G. FRIJTERS ◽  
Roelof OTTENHOFF ◽  
Michel J. A. van WIJLAND ◽  
Carin M. J. van NIEUWKERK ◽  
Albert K. GROEN ◽  
...  
Keyword(s):  
Bile Salt ◽  
Bile Salts ◽  
Control Diet ◽  
Mrna Level ◽  
Northern Blotting ◽  
Mrna Levels ◽  
Male Mice ◽  
Complete Replacement ◽  
P Glycoprotein ◽  
Mrna Content

The phosphatidyl translocating activity of the mdr2 P-glycoprotein (Pgp) in the canalicular membrane of the mouse hepatocyte is a rate-controlling step in the biliary secretion of phospholipid. Since bile salts also regulate the secretion of biliary lipids, we investigated the influence of the type of bile salt in the circulation on mdr2 Pgp expression and activity. Male mice were fed a purified diet to which either 0.1% (w/w) cholate or 0.5% (w/w) ursodeoxycholate was added. This led to a near-complete replacement of the endogenous bile salt pool (mainly tauromuricholate) by taurocholate or tauroursodeoxycholate respectively. The phospholipid secretion capacity was then determined by infusion of increasing amounts of tauroursodeoxycholate. Cholate feeding resulted in a 55% increase in maximal phospholipid secretion compared with that in mice on the control diet. Northern blotting revealed that cholate feeding increased mdr2 Pgp mRNA levels by 42%. Feeding with ursodeoxycholate did not influence the maximum rate of phospholipid output or the mdr2 mRNA content. Female mice had a higher basal mdr2 Pgp mRNA level than male mice, and this was also correlated with a higher phospholipid secretion capacity. This could be explained by the 4-fold higher basal cholate content in the bile of female compared with male mice. Our results suggest that the type of bile salts in the circulation influences the expression of the mdr2 gene.

Abstract 107: Mir-431 as a Potential Master Regulator in Angiotensin Ii-induced Vascular Injury

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Kugeng Huo ◽  
Tlili Barhoumi ◽  
Julio C Fraulob-Aquino ◽  
Chantal Richer ◽  
Mathieu Lajoie ◽  
...  
Keyword(s):  
Vascular Injury ◽  
Molecular Networks ◽  
Type I ◽  
Mirna Cluster ◽  
Mrna Levels ◽  
Ang Ii ◽  
Master Regulator ◽  
Developmental Processes ◽  
Altered Expression ◽  
Total Rna

Objective: Vascular injury is an early manifestation and a cause of end-organ damage in hypertension. microRNAs (miRNAs) play an important role in cardiovascular disease, but their implication in vascular injury is remains unclear. We aim to use RNA sequencing (seq) and a systems biology approach to identify master regulators that mediate global gene expression changes in the course of vascular injury. Methods and Results: Ten week-old male C57BL/6 mice were infused or not with angiotensin (Ang) II (1 μg/kg/min, SC) for 14 days. Blood pressure (BP) was measured by telemetry. Total RNA was extracted from the mesenteric vasculature for total RNA and small RNA-seq. Differentially expressed (DE) miRNAs (23 up and 12 down) and mRNAs (550 up and 256 down) were identified (1.5-fold, q <0.05). Molecular networks were constructed to integrate predicted interactions between DE miRNAs and inversely expressed DE mRNAs and between DE transcription factors (TF) and DE genes. Gene enrichment analysis revealed DE mRNAs involved in extracellular matrix (ECM) and developmental processes regulated by DE miRNAs ( q <1.5E-11). Seventeen upregulated miRNAs are located in the miRNA cluster of the Dlk1-Dio3 region that is highly conserved in humans, 9 of which had expression levels correlated with BP ( P <0.05). Among those 9, miR-431 that ranked first as DE miRNA ( q <0.0005) and is 100% conserved in humans, and a conserved putative DE target, a BP-correlated ( P <0.05) TF ETS homologous factor ( Ehf ), which regulates numerous ECM genes including collagen type I α1 ( Col1a1 ), were selected for functional studies. Transfection of a miR-431 mimic in human aortic smooth muscle cells (HASMCs) decreased Ehf (0.1±0.1-fold, P <0.001) and increased Ehf -suppressing target Col1a1 (1.7±0.5-fold, P <0.001) mRNA levels. Transfection of a miR-431 inhibitor caused reciprocal effects ( P <0.05). Ehf siRNA knockdown increased Col1a1 (1.2±0.1-fold, P <0.001) mRNA levels. Conclusions: Ang II infusion altered expression of miRNAs in the Dlk1-Dio3 cluster and genes involved in ECM and developmental processes. miR-431 targets TF Ehf , which leads to increased Col1a1 in HASMCs. miR-431 may act as a master regulator for vascular injury and could be a potential therapeutic target.

scholarly journals The effect of intermittent umbilical cord occlusion on insulin-like growth factors and their binding proteins in preterm and near-term ovine fetuses

2000 ◽  
Vol 166 (3) ◽  
pp. 565-577 ◽  
Author(s):  
LR Green ◽  
Y Kawagoe ◽  
DJ Hill ◽  
BS Richardson ◽  
VK Han
Keyword(s):  
Skeletal Muscle ◽  
Growth Factors ◽  
Umbilical Cord ◽  
Fetal Growth ◽  
Binding Proteins ◽  
Negative Impact ◽  
Mrna Levels ◽  
Igf I ◽  
Near Term ◽  
Insulin Like Growth Factors

Intermittent umbilical cord compression with resultant fetal hypoxia can have a negative impact on fetal growth and development. Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) are the most important regulators of fetal growth. In preterm (107-108 days of gestation) and near-term (128-131 days of gestation) ovine fetuses, we have determined the effect of intermittent umbilical cord occlusion (UCO) over a period of 4 days on the profile and expression of IGFs and IGFBPs. In experimental group animals (preterm n=7; near term n=7) UCOs were carried out by complete inflation of an occluder cuff (duration 90 s) every 30 min for 3-5 h each day, while control fetuses (preterm n=7; near term n=7) received no UCOs. Ewes were euthanized at the end of day 4, and fetal heart, lung, kidney, liver, skeletal muscle and placenta were collected. During UCOs, PO(2! ) fell (by approximately 13 mmHg), pH fell (by approximately 0.05) and PCO(2) increased (by approximately 7 mmHg), and changed to a similar extent in both preterm and near-term groups. In both preterm and near-term groups, there was no difference in fetal body or organ weight between UCO and control fetuses. No significant changes were observed in plasma IGF-I and -II concentrations or IGFBP-1, -2, -3 or -4 levels throughout the 4-day study at either gestational age. In the preterm group UCO fetuses, IGF-II mRNA (1.2-6.0 kb) levels were lower in fetal lung (33%, P<0.05), heart (54%, P<0.01) and skeletal muscle (29%, P<0.05), but there were no differences in IGF-I mRNA levels (7.3 kb); IGFBP-2 mRNA (1.5 kb) levels were lower in the right lobe of the liver (42%, P<0.05) and kidney (22%, P<0.01), but hig! her in the heart (72%, P<0.01), while IGFBP-4 (2.4 kb) levels were lower in skeletal muscle (21%, P<0.01). In the near-term group UCO fetuses, IGFBP-2 mRNA levels were greater in the placenta (39%, P<0.05). Thus, intermittent UCO as studied has a greater effect on the expression of genes encoding certain peptides of the fetal IGF system in selected tissues in preterm fetuses than that in near-term fetuses. Altered IGFBP-2 mRNA levels with reduced IGF-II mRNA levels in selected tissues may mediate changes in growth and/or differentiation that might become apparent if the length of the UCO study were extended.

scholarly journals Liver nuclear hormone receptor PPAR, LXR and RXR expression and blood lipid and glucose levels in susceptible and resistent to hepatocarcinogenesis strain of mice

2010 ◽  
Vol 56 (4) ◽  
pp. 480-489
Author(s):  
E.N. Pivovarova ◽  
N.V. Baginskaya ◽  
M.L. Perepechaeva ◽  
S.I. Ilnitskaya ◽  
M.I. Dushkin
Keyword(s):  
Tumor Development ◽  
Corticosterone Level ◽  
Blood Lipid ◽  
Nuclear Hormone Receptor ◽  
Mouse Strains ◽  
Peroxisome Proliferator ◽  
Mrna Levels ◽  
Peroxisome Proliferator Activated Receptor ◽  
Mrna Content ◽  
Dd Mice

Earlier it was shown that male mice of the DD/He strain were highly susceptible to ortho-aminoasotoluene (OAT) induced hepatocarcinogenesis, and resistant to spontaneous liver tumor development as compared to the СС57BR/Mv strain. In the present work we have made a comparative investigation of peroxisome proliferator-activated receptor (PPAR), liver X-receptor (LXR) and retinoic X-receptor (RXR) mRNA levels in liver as well as concentrations of corticosterone, glucose, lipids and insulin in blood of male DD/He and СС57BR/Mv mice. Using the multiplex RT-PCR method it was found that PPAR-α, PPAR-γ, RXR-α and RXR-β mRNA content was essentially decreased in the liver of DD mice as compared to mice of the СС57BR strain. No significant interstrain differences of LXR-α and LXR-β mRNA content were found. In DD micetere was more then the 3-fold decrease of blood content of corticosterone, which is involved in PPAR and RXR regulation. DD mice demonstrated a significant decrease in blood serum glucose and insulin concentrations as well as higher reactivity to insulin as compared with СС57BR mice. Elevated blood total cholesterol and cholesterol HDL level were found in DD mice whereas triglyceride content was basically the same in both mouse strains. It is known that glucocorticoids, PPAR and RXR play crucial role in transcription regulation of inflammation response. Therefore our data allow to suggest that decreased corticosterone level in blood, PPAR and RXR mRNA content in liver of the DD strain may lead to induction of inflammation by OAT exposure, resulting in a high incidence of tumorigenesis in this strain.

Studies of Muc-1 mucin expression and polarity in the mouse mammary gland demonstrate developmental regulation of Muc-1 glycosylation and establish the hormonal basis for mRNA expression

1992 ◽  
Vol 101 (1) ◽  
pp. 191-199 ◽  
Author(s):  
G. Parry ◽  
J. Li ◽  
J. Stubbs ◽  
M.J. Bissell ◽  
C. Schmidhauser ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Cdna Probe ◽  
Mouse Mammary Gland ◽  
Mammary Development ◽  
Mammary Epithelial ◽  
Mrna Levels ◽  
Apical Surface ◽  
Ductal Cells ◽  
Carbohydrate Epitopes ◽  
Mucin Expression

Muc-1 is a major mucin glycoprotein expressed on the surface of mammary epithelial cells. It has attracted considerable attention as it is expressed in an aberrant form on many breast tumor cells. Here we describe studies using a recently obtained cDNA probe of Muc-1 expression during lactogenic development in the mouse. Northern blot analysis demonstrated that Muc-1 is expressed at all stages of lactogenic development but its levels are increased very significantly during mid-pregnancy and into lactation. The basis of this was examined using CID-9 mammary epithelial cell cultures. It was found that in the presence of insulin Muc-1 mRNA levels were increased by both hydrocortisone and prolactin, with the combination of the three hormones supporting maximum expression. Muc-1 mRNA levels were also modulated by culturing cells on a basement-membrane-like extracellular matrix that promoted mRNA levels 5- to 10-fold above levels in cells cultured on plastic tissue culture dishes. Immunocytochemical studies using monoclonal antibodies to carbohydrate epitopes on Muc-1 demonstrated that while Muc-1 was found at all developmental stages, it became increasingly sialylated during the course of pregnancy and into lactation. Additionally, we found that while Muc-1 is tightly polarized to the apical surface of the epithelium of lactating and pregnant mice it exhibited a less-polarized distribution on a small proportion of ductal cells in virgin mice. We conclude that the expression of Muc-1 is regulated at several different levels and by a number of different factors. We speculate that this may reflect different functional roles for Muc-1 at different stages of mammary development.

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