scholarly journals The Trichohyalin-Like Protein Scaffoldin Is Expressed in the Multilayered Periderm during Development of Avian Beak and Egg Tooth

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 248
Author(s):  
Veronika Mlitz ◽  
Marcela Hermann ◽  
Maria Buchberger ◽  
Erwin Tschachler ◽  
Leopold Eckhart
Keyword(s):  
Sequence Similarity ◽  
Coturnix Japonica ◽  
Amino Acid Sequence Similarity ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Embryonic Tissues ◽  
Lower Beak ◽  
High Amino Acid ◽  
Evolutionarily Conserved ◽  
Polymerase Chain

Scaffoldin, an S100 fused-type protein (SFTP) with high amino acid sequence similarity to the mammalian hair follicle protein trichohyalin, has been identified in reptiles and birds, but its functions are not yet fully understood. Here, we investigated the expression pattern of scaffoldin and cornulin, a related SFTP, in the developing beaks of birds. We determined the mRNA levels of both SFTPs by reverse transcription polymerase chain reaction (RT-PCR) in the beak and other ectodermal tissues of chicken (Gallus gallus) and quail (Coturnix japonica) embryos. Immunohistochemical staining was performed to localize scaffoldin in tissues. Scaffoldin and cornulin were expressed in the beak and, at lower levels, in other embryonic tissues of both chickens and quails. Immunohistochemistry revealed scaffoldin in the peridermal compartment of the egg tooth, a transitory cornified protuberance (caruncle) on the upper beak which breaks the eggshell during hatching. Furthermore, scaffoldin marked a multilayered peridermal structure on the lower beak. The results of this study suggest that scaffoldin plays an evolutionarily conserved role in the development of the avian beak with a particular function in the morphogenesis of the egg tooth.

Molecular cloning, expression analysis and developmental changes in ovarian follicles of goose 3β-hydroxysteroid dehydrogenase 1

2014 ◽  
Vol 54 (8) ◽  
pp. 992 ◽  
Author(s):  
Yingying Zhang ◽  
Hehe Liu ◽  
Mingjun Yang ◽  
Shengqiang Hu ◽  
Liang Li ◽  
...  
Keyword(s):  
Adrenal Gland ◽  
Follicular Development ◽  
Hydroxysteroid Dehydrogenase ◽  
Ovarian Follicles ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Human And Mouse ◽  
Main Factor

The enzyme 3β-hydroxysteroid dehydrogenase/isomerase1 (3βHSD1) can catalyse the conversion of pregnenolone to progesterone in the △4-3-ketosteroid metabolic pathway. The aim of the present study was to clone 3βHSD1 and to determine whether this enzyme in the follicular wall has an effect on yolk progesterone in geese (Anser cygnoides). A putative coding sequence of 3βHSD1, which was 1134 nucleotides in length, was successfully obtained by using reverse transcription polymerase chain reaction (RT–PCR). A comparison of the deduced amino acid sequence with chicken, quail, zebra finch, cattle, horse, pig, human and mouse 3βHSD1 showed 89.7%, 88.4%, 87.3%, 55.6%, 54.0%, 53.5%, 55.3% and 52.9% similarity, respectively. The detection of 3βHSD1 mRNA levels in several tissues by quantitative real-time PCR showed that the highest level of 3βHSD1 was in the adrenal gland, followed by the ovary, which indicated that the gene we obtained was the adrenal gland/gonad-specific one. We measured the level of 3βHSD1 mRNA in the follicular wall and determined the concentration of progesterone in the yolk of these ovarian follicles; the concentration of progesterone in the yolk had a pattern of expression similar to that of 3βHSD1 in the follicular wall during follicular development. This result suggests that the expression of 3βHSD1 in the follicular wall may be a main factor that contributes to the accumulation of yolk progesterone.

scholarly journals pH-dependent regulation of the multi-subunit cation/proton antiporter Pha1 system from Sinorhizobium meliloti

Microbiology ◽  
2009 ◽  
Vol 155 (8) ◽  
pp. 2750-2756 ◽  
Author(s):  
Toshio Yamaguchi ◽  
Fuminori Tsutsumi ◽  
Péter Putnoky ◽  
Masahiro Fukuhara ◽  
Tatsunosuke Nakamura
Keyword(s):  
Sinorhizobium Meliloti ◽  
Sequence Similarity ◽  
Plant Root ◽  
Functional Expression ◽  
Amino Acid Sequence Similarity ◽  
Ph Optimum ◽  
Cation Selectivity ◽  
High Amino Acid ◽  
Alkaline Conditions ◽  
Ph Dependent

The pha1 gene cluster (pha1A′-G) of Sinorhizobium meliloti has previously been characterized as a necessary component for proper invasion into plant root tissue. It has been suggested to encode a multi-subunit K+/H+ antiporter, since mutations in the pha1 region rendered S. meliloti cells sensitive to K+ and alkali, and because there is high amino acid sequence similarity to previously characterized multi-subunit cation/H+ antiporters (Mrp antiporters). However, the detailed transport properties of the Pha1 system are yet to be determined. Interestingly, most of the Mrp antiporters are highly selective for Na+, unlike the Pha1 system. Here, we report the functional expression of the Pha1 system in Escherichia coli and the measurement of cation/H+ antiport activity. We showed that the Pha1 system is indeed a K+/H+ antiporter with a pH optimum under mildly alkaline conditions. Moreover, we found that the Pha1 system can transport Na+; this was unexpected based on previous phenotypic analyses of pha1 mutants. Furthermore, we demonstrated that the cation selectivity of the Pha1 system was altered when the pH was lowered from the optimum. The downregulation of Na+/H+ and K+/H+ antiport activities upon acidic shift appeared to occur via different processes, which might indicate the presence of distinct mechanisms for the regulation of the K+/H+ and Na+/H+ antiport activities of the Pha1 system.

Distribution and regulation of guanylyl cyclase type B in the rat nephron

1996 ◽  
Vol 270 (2) ◽  
pp. F311-F318 ◽  
Author(s):  
A. D. Dean ◽  
V. M. Vehaskari ◽  
D. Ritter ◽  
J. E. Greenwald
Keyword(s):  
Guanylyl Cyclase ◽  
Rat Kidney ◽  
Immunofluorescent Staining ◽  
Hydration Status ◽  
Mrna Levels ◽  
Type B ◽  
Rt Pcr ◽  
Data Link ◽  
Renal Inner Medulla ◽  
Polymerase Chain

C-type natriuretic peptide (CNP) has been localized to the proximal and distal nephron. In this study, we examined the distribution and regulation of the CNP receptor, guanylyl cyclase type B (GC-B), in the rat kidney. GC-B mRNA was detected most frequently in microdissected glomeruli, thin and thick limbs of the loop of Henle, and outer and inner medullary collecting ducts by reverse transcription-polymerase chain reaction (RT-PCR). This pattern of expression is supported by immunofluorescent staining, using anti-GC-B-specific antiserum. Nearly equivalent levels of GC-B and guanylyl cyclase type A (GC-A) mRNAs were found by quantitative RT-PCR (5,662 +/- 1,622 and 5,187 +/- 1,204 molecules of cDNA/microgram total RNA, respectively; means +/- SE, n = 6). Renal inner medulla GC-B mRNA levels, but not renal CNP mRNA levels, were 3.2-fold greater in hypervolemic and 2.3-fold less in hypovolemic rats compared with euvolemic controls. Immunohistochemical staining also supports a greater GC-B expression with increased volume status. These data link hydration status and GC-B expression and suggest an additional and novel mechanism for regulating intravascular volume.

scholarly journals Inhibition of mitogen-activated protein kinase by a Drosophila dual-specific phosphatase

2000 ◽  
Vol 349 (3) ◽  
pp. 821-828 ◽  
Author(s):  
Won-Jae LEE ◽  
Sun-Hong KIM ◽  
Yong-Sik KIM ◽  
Sung-Jun HAN ◽  
Ki-Sook PARK ◽  
...  
Keyword(s):  
Amino Acid ◽  
Protein Kinase ◽  
Expressed Sequence Tag ◽  
Mapk Pathway ◽  
Sequence Similarity ◽  
Mitogen Activated Protein Kinase ◽  
Amino Acid Sequence Similarity ◽  
Calculated Molecular Mass ◽  
High Amino Acid ◽  
Mitogen Activated Protein

The Drosophila extracellular signal-regulated kinase (DERK) mitogen-activated protein kinase (MAPK) is involved in the regulation of multiple differentiation and developmental processes. Tight control of MAPK activity is critical for normal cell behaviour. We identified a novel Drosophila MAPK phosphatase (DMKP) cDNA from the expressed-sequence-tag database and characterized it. Analysis of the nucleotide sequence revealed an open reading frame encoding the 203-amino acid protein, with a calculated molecular mass of 23kDa, which has a high amino acid sequence similarity with ‘VH1-like’dual-specific phosphatases at the broad region near the catalytic sites. The expression of DMKP mRNA occurs from the late larval stages to adulthood in Drosophila development. The recombinant DMKP protein produced in yeast retained its phosphatase activity. When expressed in Schneider cells, DMKP dose-dependently inhibited DERK and Drosophila c-Jun N-terminal kinase activities with high selectivity towards DERK. However, DMKP did not have any affect on Drosophila p38 activity. When DMKP was expressed in yeast, it down-regulated the fus1-lacZ trans-reporter gene of the pheromone MAPK pathway without any significant effect on the high-osmolarity-glycerol-response pathway.

scholarly journals Ontogeny of erythropoietin gene expression in the sheep fetus: effect of dexamethasone at 60 days of gestation

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 460-466
Author(s):  
GB Lim ◽  
K Jeyaseelan ◽  
EM Wintour
Keyword(s):  
Gene Expression ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Fetal Plasma ◽  
Liver And Kidney ◽  
Polymerase Chain ◽  
High Level ◽  
A Site ◽  
Sheep Fetus

We have used competitive reverse transcription and polymerase chain reaction (RT/PCR) to compare the levels of erythropoietin (Epo) mRNA in the liver and kidneys of the sheep fetus at 60, 80, 100, 130, and 140 days of gestation (term = 145 to 150 days). The effect of dexamethasone infusion in the ewe on Epo gene expression in the 60-day fetus was also investigated. Epo mRNA levels were highest at 60 days of gestation, the earliest age studied, in both liver and kidney. In the liver, Epo mRNA expression declined as gestation proceeded. Kidney Epo mRNA was maintained at a high level until 100 days of gestation, declining significantly in the 130-day fetus (P < .01). Treatment of ewes carrying 60-day fetuses with 0.76 mg/h dexamethasone for 48 hours resulted in a significant decrease in fetal plasma Epo values and Epo mRNA levels in both the liver and kidney. In the dexamethasone-treated fetuses, Epo mRNA in the liver was 52% of control values (P < .05), and in the kidney, 33% of control (P < .001). The results suggest that the kidney may play a more important role as a site of Epo synthesis in the early gestation sheep fetus than previously thought. Glucocorticoids may have a role in the regulation of Epo gene expression.

Beta 3-adrenoceptor in guinea pig brown and white adipocytes: low expression and lack of function

1996 ◽  
Vol 271 (6) ◽  
pp. R1729-R1738 ◽  
Author(s):  
C. Atgie ◽  
G. Tavernier ◽  
F. D'Allaire ◽  
T. Bengtsson ◽  
L. Marti ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Cold Acclimation ◽  
Thermal Environment ◽  
Uncoupling Protein ◽  
Sequence Similarity ◽  
Amino Acid Sequence Similarity ◽  
Mrna Levels ◽  
Fat Depots ◽  
Specific Regulation ◽  
A Site

In the guinea pig, cold acclimation induced a conversion of unilocular to multilocular adipocytes in interscapular (IS) and retroperitoneal (RP) fat depots but not in the epididymal (EP) fat pad. The conversion was associated with an increase in mitochondriogenesis and the appearance of the uncoupling protein. The maximal lipolytic responses to norepinephrine and dibutyryl adenosine 3',5'-cyclic monophosphate were decreased in IS cells, unchanged in RP cells, and increased in EP cells, suggesting a site-specific regulation of lipolysis at the postreceptor level. beta 3-Adrenergic agonists were not lipolytic regardless of the depot and the thermal environment of the animal. These agents did not inhibit glucose transport and lipogenesis, as was previously reported for rodents. Cloning and sequencing of the guinea pig beta 3-adrenoceptor gene revealed a slightly higher amino acid sequence similarity with the human than with the rodent beta 3-adrenoceptors. beta 3-Adrenoceptor transcripts were present at a very low level in guinea pig adipocytes, and mRNA levels did not increase to a significant extent after cold acclimation. The guinea pig thus differs from rodents by an absence of beta 3-adrenergic effects and by low beta 3-adrenoceptor expression in brown and white adipose tissues.

UL 16 Binding Protein 3 Gene Expression: Does It Have an Association With Alopecia Areata?

2020 ◽  
Author(s):  
Rania Azmy ◽  
Alaa Maraee ◽  
Eman Amer ◽  
Nermin Tayel ◽  
Wafaa Ahmed Shehata
Keyword(s):  
Gene Expression ◽  
Alopecia Areata ◽  
Disease Duration ◽  
Binding Protein ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Polymerase Chain ◽  
Insight Into ◽  
Gene Expression Levels ◽  
Gaining Insight

Abstract ObjectiveAlopecia Areata is one of the most widespread autoimmune diseases affecting both sexes of all ages and across all ethnic individuals. Genetics is considered to be a valuable tool for gaining insight into the disease’s pathogenesis. Association with UL 16 Binding Protein (ULBP) genes has been detected with autoimmune disorders.This study aimed to detect UL-16 Binding Protein -3 (ULBP3) gene expression levels in cases with AA and to correlate those levels with the clinical course of the disease.This study included 85 subjects: 55 patients with AA and 30 age- and sex-matched healthy controls. The expression level of ULBP3 mRNA was estimated using Real-Time Polymerase Chain Reaction (RT-PCR).Results Levels of ULBP3 mRNA in cases were significantly higher in patients with AA in comparison with controls. Also, there were significant correlations between ULBP3 mRNA levels and age of patients and disease duration in years. ULBP3 upregulation in AA enforces the theory that postulates the autoimmune nature of AA and ULBP3 may be involved in AA pathogenesis and its progression.

Influence of mechanical and biological signals on gene expression in human MG-63 cells: evidence for a complex interplay between hydrostatic compression and vitamin D3 or TGF-β1 on MMP-1 and MMP-3 mRNA levels

2005 ◽  
Vol 83 (1) ◽  
pp. 96-107 ◽  
Author(s):  
V Tasevski ◽  
J M Sorbetti ◽  
S S Chiu ◽  
N G Shrive ◽  
D A Hart
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Polymerase Chain Reaction ◽  
Bone Cells ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tgf Β1 ◽  
In Cells

Biological mediators can influence the activity and differentiation of bone cells. 1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3) is known to induce differentiation of precursors into mature osteoblasts, and transforming growth factorβ1 (TGF-β1) can modulate the activity of bone cells leading to alterations in proliferation and gene expression patterns. Bone-derived cells were loaded via intermittent cyclic hydrostatic pressure (icHP) on cells under basal conditions and in the presence of 1,25-(OH)2D3 or TGF-β1. Evaluating the effects of loading on the cells allowed for a comparison to be made between responsiveness to biomechanical and biochemical stimuli and their potential interplay. The effects of icHP on mRNA levels for the specific genes involved in bone remodelling and differentiation were measured in MG-63 cells using reverse transcription-polymerase chain reaction (RT-PCR). The mRNA levels for matrix metalloproteinase-1 and -3 (MMP-1 and MMP-3) were significantly, and uniquely, increased (p < 0.001) in cells exposed to icHP under serum-free conditions for 4–12 h. However, mRNA levels for MMP-3, but not MMP-1, were significantly enhanced in cells subjected to static hydrostatic pressure (HP). Treatment of cells with 1,25-(OH)2D3 resulted in increased (p < 0.001) mRNA levels for osteocalcin and decreased (p < 0.001) mRNA levels for both MMP-1 and MMP-3. In cells exposed to icHP and 1,25-(OH)2D3, the mRNA levels for both MMP-1 and MMP-3 were elevated (p < 0.001) compared with hormone alone, but not to the same degree (p < 0.01) as cells subjected to icHP alone. Addition of TGF-β1 to cells led to increases in cell proliferation and expression of collagen I, as well as decreases in expression of osteocalcin and MMP-1 and MMP-3. Exposure of cells to icHP and TGF-β1 again led to unique and significant increases in expression of MMP-1 and MMP-3. No changes in mRNA levels for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or any of the other 9 genes assessed, including those for MMP-2 and MMP-13, were detected under any of the conditions described. Therefore, icHP can induce alterations in mRNA levels for a specific subset of genes in both premature and mature osteoblasts. Such stimuli can modulate the impact of potent biological mediators in defining patterns of gene expression by bone cells and potentially modify function in vivo.Key words: osteoblast, biomechanical loading,1,25-dihydroxy-vitamin D3 (1,25-(OH)2D3), mRNA levels, reverse trans cription-polymerase chain reaction (RT-PCR), transforming growth factor-β1 (TGF-β1).

scholarly journals Yes-associated protein (YAP) promotes progression of glioma and poor prognosis of human

2021 ◽  
Author(s):  
Fei Yan ◽  
Lele Du ◽  
Jiatao Lv ◽  
Haitao Zhang ◽  
Jianxin Zhu ◽  
...  
Keyword(s):  
Malignant Glioma ◽  
Brain Tissue ◽  
Malignant Tumors ◽  
Normal Brain ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Glioma Tissue ◽  
Normal Brain Tissue ◽  
Polymerase Chain ◽  
Relationship Of

Abstract Background: Yes-associated protein(YAP) plays an important role in signal transduction and gene transcription regulation in 1 normal cells, with elevated and over-expressed YAP levels observed in various malignant tumors. The aim of this study was 2 to investigate the expression of YAP in malignant glioma, and to study the possible relationship of YAP expression with the 3 occurrence and development of malignant glioma. 4 Methods: Immunohistochemical staining was used to assess the expression of YAP and phosphor-YAP in malignant glioma 5 tissue and normal brain tissue, and their protein and mRNA levels were evaluated through Western blotting and reverse 6 transcription-polymerase chain reaction (RT-PCR), respectively. Normal brain tissue obtained from the functional lesion of 7 the epilepsy patients. After transfection of YAPsiRNA oligonucleotides or pcDNA3.1-hYAP plasmid, their effects on glioma 8 cells were investigated using western blot, cell proliferation, cycle, apoptosis and invasion, respectively. We conducted the 9 2 co-Immunoprecipitation to verify the combination of YAP and PPARγ, explore the mechanism of action. 10 Results: YAP-positive expression was found in 9 cases of normal brain and 60 cases of glioma. A significantly higher 11 expression of YAP in glioma tissue as compared with normal brain tissue at both protein and mRNA levels, and YAP proteins 12 mainly expressed and located in the nucleus and only a small percentage in the cytoplasm of glioma tissue. Phosphor-YAP 13 protein expression showed high staining of the cytoplasm, but no staining of the nuclear. While, with the enhancement of 14 the malignant degree, the cytoplasm YAP(p-YAP) expression is lower gradually than normal brain tissues. Further study in 15 glioma cell lines in which YAP was either overexpressed or depleted confirmed that YAP markedly promoted the cell 16 proliferation, cycle, invasion and inhibited the cell apoptosis. Moreover, YAP in company with PPARγ regulates the cell 17 proliferatin and effects the gliomagenesis. 18 Conclusion: These results indicate that YAP plays an important role in glioma and might be a useful therapeutic target of 19 glioma. 20

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