Identification and characterization of seven new exon 11-associated splice variants of the rat mu opioid receptor gene, OPRM1

<p>Abstract</p> <p>Background</p> <p>The mouse mu opioid receptor (OPRM1) gene undergoes extensive alternative splicing at both the 3'- and 5'-ends of the gene. Previously, several C-terminal variants generated through 3' splicing have been identified in the rat OPRM1 gene. In both mice and humans 5' splicing generates a number of exon 11-containing variants. Studies in an exon 11 knockout mouse suggest the functional importance of these exon 11-associated variants in mediating the analgesic actions of a subset of mu opioids, including morphine-6β-glucuronide (M6G) and heroin, but not others such as morphine and methadone. We now have examined 5' splicing in the rat.</p> <p>Results</p> <p>The current studies identified in the rat a homologous exon 11 and seven exon 11-associated variants, suggesting conservation of exon 11 and its associated variants among mouse, rat and human. RT-PCR revealed marked differences in the expression of these variants across several brain regions, implying region-specific mRNA processing of the exon 11-associated variants. Of the seven rat exon 11-associated variants, four encoded the identical protein as found in rMOR-1, two predicted 6 TM variants, and one, rMOR-1H2, generated a novel N-terminal variant in which a stretch of an additional 50 amino acids was present at the N-terminus of the previously established rMOR-1 sequence. When expressed in CHO cells, the presence of the additional 50 amino acids in rMOR-1H2 significantly altered agonist-induced G protein activation with little effect on opioid binding.</p> <p>Conclusion</p> <p>The identification of the rat exon 11 and its associated variants further demonstrated conservation of 5' splicing in OPRM1 genes among rodents and humans. The functional relevance of these exon 11 associated variants was suggested by the region-specific expression of their mRNAs and the influence of the N-terminal sequence on agonist-induced G protein coupling in the novel N-terminal variant, rMOR-1H2. The importance of the exon 11-associated variants in mice in M6G and heroin analgesia revealed in the exon 11 knockout mouse implies that these analogous rat variants may also play similar roles in rat. The complexity created by alternative splicing of the rat OPRM1 gene may provide important insights of understanding the diverse responses to the various mu opioids seen in rats.</p

Identification and characterization of seven new exon 11-associated splice variants of the rat mu opioid receptor gene, OPRM1

<p>Abstract</p> <p>Background</p> <p>The mouse mu opioid receptor (OPRM1) gene undergoes extensive alternative splicing at both the 3'- and 5'-ends of the gene. Previously, several C-terminal variants generated through 3' splicing have been identified in the rat OPRM1 gene. In both mice and humans 5' splicing generates a number of exon 11-containing variants. Studies in an exon 11 knockout mouse suggest the functional importance of these exon 11-associated variants in mediating the analgesic actions of a subset of mu opioids, including morphine-6β-glucuronide (M6G) and heroin, but not others such as morphine and methadone. We now have examined 5' splicing in the rat.</p> <p>Results</p> <p>The current studies identified in the rat a homologous exon 11 and seven exon 11-associated variants, suggesting conservation of exon 11 and its associated variants among mouse, rat and human. RT-PCR revealed marked differences in the expression of these variants across several brain regions, implying region-specific mRNA processing of the exon 11-associated variants. Of the seven rat exon 11-associated variants, four encoded the identical protein as found in rMOR-1, two predicted 6 TM variants, and one, rMOR-1H2, generated a novel N-terminal variant in which a stretch of an additional 50 amino acids was present at the N-terminus of the previously established rMOR-1 sequence. When expressed in CHO cells, the presence of the additional 50 amino acids in rMOR-1H2 significantly altered agonist-induced G protein activation with little effect on opioid binding.</p> <p>Conclusion</p> <p>The identification of the rat exon 11 and its associated variants further demonstrated conservation of 5' splicing in OPRM1 genes among rodents and humans. The functional relevance of these exon 11 associated variants was suggested by the region-specific expression of their mRNAs and the influence of the N-terminal sequence on agonist-induced G protein coupling in the novel N-terminal variant, rMOR-1H2. The importance of the exon 11-associated variants in mice in M6G and heroin analgesia revealed in the exon 11 knockout mouse implies that these analogous rat variants may also play similar roles in rat. The complexity created by alternative splicing of the rat OPRM1 gene may provide important insights of understanding the diverse responses to the various mu opioids seen in rats.</p

Effects of a Chinese Medicinal Plant Radix Astragali on the Ovariectomized Female Rats

Perimenopausal syndrome occurs during the transition to menopause. Complementary and alternative medicine, especially Chinese medicinal plants, has manifested significant effects in alleviating perimenopausal symptoms. However, little research has been focused on the effects of Chinese medicinal plant on the immune function of the perimenopausal women. The present study aimed to explore the effects of Radix Astragali (RA) on the sex hormone levels and the interleukins of the ovariectomized female rats. 24 female Sprague-Dawley (SD) rats were randomly divided into model control group (MOD group), sham-operation group (SHAM group), RA group and estrogen group (EST group). After all the treatment ended, the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), interleukin-2 (IL-2), and interleukin-8 (IL-8) were measured using enzyme-linked immune-sorbent assay (ELISA) and the uterus was removed and weighed after blood exsanguinations immediately. In the MOD group, the serum levels of E2 were significantly lower, and the serum levels of FSH and LH were markedly higher than those of the RA group, EST group and SHAM group (P&lt;0.05). In the RA group, the serum levels of E2 were significantly lower, and the serum levels of FSH were markedly higher than those of the SHAM group and EST group, respectively. In the MOD group, the serum levels of IL-2 and IL-8 were significantly lower than those of the RA group, EST group and SHAM group (P&lt;0.05), and no marked differences existed among RA group, EST group and SHAM group in the serum levels of IL-2 and IL-8 (P&gt;0.05). The uterine weight of the rats in the RA group, EST group and SHAM group were significantly higher than those of the rats in MOD group (P&lt;0.05). There were no marked differences among the rats from RA group, EST group and SHAM group on the uterine weight (P&gt;0.05). It is concluded that RA can significantly improve the immune functions of the ovariectomized female rats, although it cannot change the sex hormones levels as significantly as estrogen

Analysis of critical risk factors for ppp sewage treatment projects based on a sewage treatment project in quanzhou

2008-2009 > Academic research: refereed > Publication in refereed journalVersion of RecordPublishe

Neural Network-Based Retinal Nerve Fiber Layer Profile Compensation for Glaucoma Diagnosis in Myopia: Model Development and Validation

BACKGROUND: Due to the axial elongation-associated changes in the optic nerve and retina in high myopia, traditional methods like optic disc evaluation and visual field are not able to correctly differentiate glaucomatous lesions. It has been clinically challenging to detect glaucoma in highly myopic eyes. OBJECTIVE: This study aimed to develop a neural network to adjust for the dependence of the peripapillary retinal nerve fiber layer (RNFL) thickness (RNFLT) profile on age, gender, and ocular biometric parameters and to evaluate the network's performance for glaucoma diagnosis, especially in high myopia. METHODS: RNFLT with 768 points on the circumferential 3.4-mm scan was measured using spectral-domain optical coherence tomography. A fully connected network and a radial basis function network were trained for vertical (scaling) and horizontal (shift) transformation of the RNFLT profile with adjustment for age, axial length (AL), disc-fovea angle, and distance in a test group of 2223 nonglaucomatous eyes. The performance of RNFLT compensation was evaluated in an independent group of 254 glaucoma patients and 254 nonglaucomatous participants. RESULTS: By applying the RNFL compensation algorithm, the area under the receiver operating characteristic curve for detecting glaucoma increased from 0.70 to 0.84, from 0.75 to 0.89, from 0.77 to 0.89, and from 0.78 to 0.87 for eyes in the highest 10% percentile subgroup of the AL distribution (mean 26.0, SD 0.9 mm), highest 20% percentile subgroup of the AL distribution (mean 25.3, SD 1.0 mm), highest 30% percentile subgroup of the AL distribution (mean 24.9, SD 1.0 mm), and any AL (mean 23.5, SD 1.2 mm), respectively, in comparison with unadjusted RNFLT. The difference between uncompensated and compensated RNFLT values increased with longer axial length, with enlargement of 19.8%, 18.9%, 16.2%, and 11.3% in the highest 10% percentile subgroup, highest 20% percentile subgroup, highest 30% percentile subgroup, and all eyes, respectively. CONCLUSIONS: In a population-based study sample, an algorithm-based adjustment for age, gender, and ocular biometric parameters improved the diagnostic precision of the RNFLT profile for glaucoma detection particularly in myopic and highly myopic eyes

Properties of plasmoids observed in Saturn’s dayside and nightside magnetodisc

Plasmoid is a key structure for transferring magnetic flux and plasma in planetary magnetospheres. At Earth, plasmoids are key media which transfer energy and mass in the "Dungey Cycle". For giant planets, plasmoids are primarily generated by the dynamic processes associated with “Vasyliunas Cycle”. It is generally believed that planetary magnetotails are favorable for producing plasmoids. Nevertheless, recent studies reveal that magnetic field lines could be sufficiently stretched to allow magnetic reconnection in Saturn’s dayside magnetodisc. In the study, we report direct observations of plasmoids in Saturn’s dayside magnetodisc for the first time. Moreover, we perform a statistical investigation on the global plasmoid electron density distribution. The results show an inverse correlation between the nightside plasmoid electron density and local time, and the maximum plasmoid electron density around prenoon local time on the dayside. These results are consistent with the magnetospheric circulation picture associated with the "Vasyliunas Cycle"

Characterization of the Major Histocompatibility Complex Class II Genes in Miiuy Croaker

Major histocompatibility complex (MHC) has a central role in the adaptive immune system by presenting foreign peptide to the T-cell receptor. In order to study the molecular function and genomic characteristic of class II genes in teleost, the full lengths of MHC class IIA and IIB cDNA and genomic sequence were cloned from miiuy croaker (Miichthys miiuy). As in other teleost, four exons and three introns were identified in miiuy croaker class IIA gene; but the difference is that six exons and five introns were identified in the miiuy croaker class IIB gene. The deduced amino acid sequence of class IIA and class IIB had 26.3–85.7% and 11.0–88.8% identity with those of mammal and teleost, respectively. Real-time quantitative RT-PCR demonstrated that the MHC class IIA and IIB were ubiquitously expressed in ten normal tissues; expression levels of MHC genes were found first upregulated and then downregulated, and finally by a recovery to normal level throughout the pathogenic bacteria infection process. In addition, we report on the underlying mechanism that maintains sequences diversity among many fish species. A series of site-model tests implemented in the CODEML program revealed that positive Darwinian selection is likely the cause of the molecular evolution in the fish MHC class II genes

Influence of magnetic disorders on quantum anomalous Hall effect in magnetic topological insulator films beyond the two-dimensional limit

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An environmentally friendly solution-processed ZrLaO gate dielectric for large-area applications in the harsh radiation environment

In this work, an eco-friendly aqueous solution-processed ZrLaO dielectric is demonstrated for large-area application in the harsh radiation environment. Appropriate La doping (10% La) into ZrOx could suppress the formation of Vo and improve the InOx/ZrLaO interface. The Zr0.9La0.1Oy thin films remained stable under 144 krad (SiO2) gamma-ray irradiation, no distinct composition variation or property degradation were observed. The resistor-loaded inverter based on InOx/Zr0.9La0.1Oy TFT demonstrated full swing characteristics with a gain of 13.3 at 4 V and remained 91% gain after 103 krad (SiO2) irradiation