Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle

1993 ◽  
Vol 265 (5) ◽  
pp. H1501-H1509 ◽  
Author(s):  
P. Ping ◽  
J. E. Faber
Keyword(s):  
Smooth Muscle ◽  
Messenger Rna ◽  
Vena Cava ◽  
Pcr Analysis ◽  
Relative Distribution ◽  
Alpha Adrenoceptor ◽  
Aortic Media ◽  
Unique Restriction ◽  
Polymerase Chain

Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR-amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Characterization of Sterol Demethylation Inhibitor-Resistant Isolates of Fusarium asiaticum and F. graminearum Collected from Wheat in China

2009 ◽  
Vol 99 (5) ◽  
pp. 487-497 ◽  
Author(s):  
Y. Yin ◽  
X. Liu ◽  
B. Li ◽  
Z. Ma
Keyword(s):  
Phytopathogenic Fungi ◽  
Cross Resistance ◽  
Pcr Analysis ◽  
Head Blight ◽  
Fusarium Asiaticum ◽  
Homologous Genes ◽  
Sterol Demethylation Inhibitors ◽  
Polymerase Chain ◽  
Dmi Resistance

Fusarium asiaticum and F. graminearum are the primary causal agents of Fusarium head blight (FHB) of wheat in China. In this study, sensitivities of 159 F. asiaticum and F. graminearum isolates to a benzimidazole fungicide carbendazim (MBC) and to sterol demethylation inhibitors (DMIs) tebuconazole and prochloraz were determined. Among the 159 isolates, 9 were resistant to MBC and designated as MBC-R isolates. Three showed resistance to tebuconazole and prochloraz and designated as DMI-R isolates. There was no cross-resistance between MBC and DMI. Genetic analysis by microsatellite-primed polymerase chain reaction (PCR) showed that MBC-R or DMI-R isolates had different genotypes, which indicated that they originated from different wild-type parents. Analysis of two 14α-demethylase (cyp51) homologous genes (cyp51A and cyp51B) showed that the F. asiaticum isolates could be distinguished from F. graminearum isolates based on the sequence of cyp51A. Analysis of deduced amino acid sequence of cyp51A and cyp51B suggested that no mutations were associated with DMI resistance. Real-time PCR analysis showed that the DMI resistance was not related to the expression of cyp51A and cyp51B in F. asiaticum and F. graminearum, but expressions of both genes were induced greatly by the tebuconazole. Results of this study indicated that cyp51A would be an informative marker for analysis of population structure of F. asiaticum and F. graminearum, and the existence of homologous cyp51 genes in F. asiaticum and F. graminearum could provide new insights into DMI resistance in phytopathogenic fungi.

Sequential opening of IP3-sensitive Ca2+channels and SOC during α-adrenergic activation of rabbit vena cava

2002 ◽  
Vol 282 (5) ◽  
pp. H1768-H1777 ◽  
Author(s):  
Cheng-Han Lee ◽  
Roshanak Rahimian ◽  
Tania Szado ◽  
Jasmin Sandhu ◽  
Damon Poburko ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Transient Receptor Potential ◽  
Vena Cava ◽  
Receptor Potential ◽  
Selective Inhibition ◽  
Pcr Analysis ◽  
Release Channel ◽  
Plasmic Reticulum ◽  
Intracellular Ca ◽  
Transient Receptor

α1-Aderenoceptor-mediated constriction of rabbit inferior vena cava (IVC) is signaled by asynchronous wavelike Ca2+ oscillations in the in situ smooth muscle. We have shown previously that a putative nonselective cationic channel (NSCC) is required for these oscillations. In this report, we show that the application of 2-aminoethoxyphenyl borate (2-APB) to antagonize inositol 1,4,5-trisphosphate (InsP3)-sensitive Ca2+ release channels (IP3R channels) can prevent the initiation and abolish ongoing α1-aderenoceptor-mediated tonic constriction of the venous smooth muscle by inhibiting the generation of these intracellular Ca2+ concentration ([Ca2+]i) oscillations. The observed effects of 2-APB can only be attributed to its selective inhibition on the IP3R channels, not to its slight inhibition of the L-type voltage-gated Ca2+ channel and the sarco(endo)plasmic reticulum Ca2+ ATPase. Furthermore, 2-APB had no effect on the ryanodine-sensitive Ca2+ release channel and the store-operated channel (SOC) in the IVC. These results indicate that the putative NSCC involved in refilling the sarcoplasmic reticulum (SR) and maintaining the tonic contraction is most likely an SOC-type channel because it appears to be activated by IP3R-channel-mediated SR Ca2+ release or store depletion. This is in accordance with its sensitivity to Ni2+ and La3+ (SOC blockers). More interestingly, RT-PCR analysis indicates that transient receptor potential (Trp1) mRNA is strongly expressed in the rabbit IVC. The Trp1 gene is known to encode a component of the store-operated NSCC. These new data suggest that the activation of both the IP3R channels and the SOC are required for PE-mediated [Ca2+]i oscillations and constriction of the rabbit IVC.

scholarly journals Molecular Cloning and Characterization of the HOS1 Gene from ‘Muscat Hamburg’ Grapevine

2014 ◽  
Vol 139 (1) ◽  
pp. 54-62 ◽  
Author(s):  
Jitao Li ◽  
Nian Wang ◽  
Lina Wang ◽  
Haiping Xin ◽  
Shaohua Li
Keyword(s):  
Polymerase Chain Reaction ◽  
Arabidopsis Thaliana ◽  
Abscisic Acid ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
The World ◽  
Drought Conditions ◽  
Polymerase Chain ◽  
Polyethylene Glycol 6000

Cold stress is an important factor that limits grape (Vitis sp.) production around the world. The high expression of osmotically responsive genes 1 (HOS1) protein acts as a repressor of cold-responsive genes in plants. To increase understanding of mechanism regulating cold tolerance in grape, we isolated and characterized a novel HOS1 gene, designated VvHOS1 from ‘Muscat Hamburg’ grapevine (Vitis vinifera). Real-time polymerase chain reaction (PCR) analysis revealed that the expression of VvHOS1 could be induced by the application of exogenous abscisic acid and various abiotic environmental conditions such as low temperature, drought, and salinity. Moreover, VvHOS1 expression could also be induced by cold plus drought conditions (4 °C, 10% polyethylene glycol 6000). In addition, overexpression of VvHOS1 in arabidopsis (Arabidopsis thaliana) decreased the plants’ tolerance to cold, drought, and salt as well as negatively regulated the expression level of two stress-responsive genes, AtRD29A and AtCOR47. The results obtained in this study should help us to elucidate the function of VvHOS1 and understand the cold-responsive pathway in grapevine.

scholarly journals Characterization of a Gene Identified in Pathotype 5 of the Clubroot Pathogen Plasmodiophora brassicae

2015 ◽  
Vol 105 (6) ◽  
pp. 764-770 ◽  
Author(s):  
H. Zhang ◽  
J. Feng ◽  
V. P. Manolii ◽  
S. E. Strelkov ◽  
S.-F. Hwang
Keyword(s):  
Plasmodiophora Brassicae ◽  
Blot Hybridization ◽  
Field Isolate ◽  
Pcr Analysis ◽  
In Planta ◽  
Dot Blot ◽  
Reverse Transcription Quantitative Pcr ◽  
Polymerase Chain ◽  
Differential Hosts

Clubroot caused by Plasmodiophora brassicae is an important disease of crucifers worldwide. Isolates of the pathogen can be classified into pathotypes according to their pathogenicity on differential hosts. In this study, the presence or absence of all database-available nonhousekeeping P. brassicae genes (118 in total) were assessed by polymerase chain reaction (PCR) analysis in isolates belonging to five P. brassicae pathotypes (2, 3, 5, 6, and 8 according to Williams’ differential set). One gene, designated Cr811, was present exclusively in the isolate of pathotype 5. This was further confirmed by dot blot hybridization and by PCR using alternative DNA preparations and primers. Reverse transcription quantitative PCR analysis indicated that in planta expression of Cr811 was up-regulated during canola infection, especially in the stage of secondary plasmodia. Primers specific to Cr811 could distinguish a field isolate of P. brassicae belonging to pathotype 5 from two other field isolates representing pathotypes 3 and 8. These findings suggest that Cr811 is a gene that is potentially involved in clubroot pathogenesis and that it also might serve as a molecular marker for differentiation of pathotype 5 from other pathotypes.

Morphological and biochemical characterization of remodeling in aorta and vena cava of DOCA-salt hypertensive rats

2007 ◽  
Vol 292 (5) ◽  
pp. H2438-H2448 ◽  
Author(s):  
Stephanie W. Watts ◽  
Catherine Rondelli ◽  
Keshari Thakali ◽  
Xiaopeng Li ◽  
Bruce Uhal ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Remodeling ◽  
Biochemical Characterization ◽  
Vena Cava ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Arterial Remodeling ◽  
Hypertensive Rats ◽  
Salt Hypertension ◽  
Expression And Activity

Arterial remodeling occurs in response to mechanical and neurohumoral stimuli. We hypothesized that veins, which are not exposed to higher pressures in hypertension, would demonstrate less active remodeling than arteries. We assessed remodeling with two standard measures of arterial remodeling: vessel morphometry and the expression/function of matrix metalloproteinases (MMPs). Thoracic aorta and vena cava from sham normotensive and DOCA-salt hypertensive rats (110 ± 4 and 188 ± 8 mmHg systolic blood pressure, respectively) were used. Wall thickness was increased in DOCA-salt vs. sham aorta (301 ± 23 vs. 218 ± 14 μm, P < 0.05), as was medial area, but neither measure was altered in the vena cava. The aorta and vena cava expressed the gelatinases MMP-2, MMP-9, transmembrane proteinase MT1-MMP, and tissue inhibitor of metalloproteinase-2 (TIMP-2). Immunohistochemically, MMP-2 localized to smooth muscle in the aorta and densely in endothelium/smooth muscle of the vena cava. Western and zymographic analyses verified that MMP-2 was active in all vessels and less active in the vena cava than aorta. In hypertension, MMP-2 expression and activity in the aorta were increased (59.1 ± 3.7 and 74.5 ± 6.1 units in sham and DOCA, respectively, P < 0.05); similar elevations were not observed in the vena cava. MMP-9 was weakly expressed in all vessels. MT1-MMP was expressed by the aorta and vena cava and elevated in the vena cava from DOCA-salt rats. TIMP-2 expression was significantly increased in the aorta of DOCA rats compared with sham but was barely detectable in the vena cava of sham or DOCA-salt hypertensive rats. These findings suggest that large veins may not undergo vascular remodeling in DOCA-salt hypertension.

scholarly journals Identification and characterization of Edwardsiella ictaluri from diseased Pangasius pangasius, cultured in Cirata Lake, Indonesia

2018 ◽  
Vol 19 (3) ◽  
pp. 816-822 ◽  
Author(s):  
MIRA MAWARDI ◽  
JAELANI JAELANI ◽  
ZAKKI ZAINUN ◽  
YUANI MUNDAYANA ◽  
BAIRD SAM CHILORA ◽  
...  
Keyword(s):  
Edwardsiella Ictaluri ◽  
Pcr Analysis ◽  
Nucleotide Sequencing ◽  
Infection Prevalence ◽  
Chain Reaction ◽  
Prevalence Level ◽  
Conventional Biochemical Test ◽  
Polymerase Chain ◽  
Identification And Characterization

Mawardi M, Jaelani, Zainun Z, Mundayana Y, Chilora BS, Hardi EH. 2018. Identification and characterization of Edwardsiella ictaluri from diseased Pangasius pangasius, cultured in Cirata Lake, Indonesia. Biodiversitas 19: 766-772. In January 2016, there were reported extensive mortalities of Pangasius pangasius cultured on cage, in Cirata Lake. Edwardsiella ictaluri is primarily recognized as a disease-causing pathogen in catfish species, causing an economically important bacterial infection hence affecting productivity of aquaculture enterprises in many regions across the E. ictaluri. The samples were 17 fishes collected from organs containing spleen, kidney, and liver. The total samples were 51 organs of fishes. The high bacteria infection prevalence level was found in kidney (29.41%). The gross pathological signs of sample of organs were abnormal sizes and abnormal color including white nodules. Bacteria were identified by conventional biochemical test, polymerase chain reaction (PCR) analysis and nucleotide sequencing. The results showed that P. pangasius cultured in cages in Cirata Lake is positively infected by E. ictaluri strain (accession number KF9071291). Other infection cases by Aeromonas sp. and Plesiomonas shigelloides as co-infection bacteria in P. pangasius were also found.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

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