Integration of qRT-PCR and Immunohistochemical Techniques for mRNA Expression and Localization of m1AChR in the Brain of Aging Rat

3-Hydroxyl-3-methylglutaryl coenzyme A reductase inhibitors (statins) have been shown to reduce the incidence of myocardial infarction independent of their lipid-lowering effects. Nitric oxide (NO) in the central nervous system contributes to cardiovascular regulatory mechanisms. Imbalance between nitric oxide (NO) and superoxide anion (O 2 . − ) in the brain may contribute to enhanced sympathetic drive in heart failure (HF). This study was done to determine whether treatment with atorvastatin (ATS) ameliorates the imbalance between NO and O 2 . − production in the brain stem and contributes to improvement of left ventricular (LV) function. Methods and Results: Myocardial infarction (MI) was induced by ligation of the left coronary artery or sham surgery. Subsequently, mice were treated with ATS (10 μg/kg) (MI + ATS), or vehicle (MI + V). After 5 weeks, echocardiography revealed left ventricular dilatation in MI mice. Realtime RT-PCR indicated an increase in the mRNA expression of the LV hypertrophy markers, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). Neuronal NOS (nNOS) and endothelial NOS (eNOS) mRNA expression were significantly reduced, while that of NAD(P)H oxidase subunit (gp91phox) expression was elevated in the brain stem of MI mice. Compared with sham-operated mice, ATS-treated mice showed reduced cardiac dilatation, decreased ANP and BNP in the LV. ATS also reduced gp91phox expression and increased nNOS mRNA expression in the brain stem, while no changes in eNOS and iNOS were observed. Conclusion: These findings suggest that ATS reduces oxidative stress and increases neuronal NOS in the brain stem, and improves left ventricular function in heart failure.

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Cloning of the rat Gadd45 cDNA and its mRNA expression in the brain

Gene ◽

10.1016/0378-1119(94)90666-1 ◽

1994 ◽

Vol 151 (1-2) ◽

pp. 253-255 ◽

Cited By ~ 18

Author(s):

Toru Yoshida ◽

Edward L. Schneider ◽

Nozomu Mori

Keyword(s):

Mrna Expression ◽

The Brain

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Neonatal phencyclidine treatment selectively attenuates mesolimbic dopamine function in adult rats as revealed by methamphetamine-induced behavior andc-fos mRNA expression in the brain

Synapse ◽

10.1002/1098-2396(200104)40:1<11::aid-syn1021>3.0.co;2-i ◽

2001 ◽

Vol 40 (1) ◽

pp. 11-18 ◽

Cited By ~ 8

Author(s):

Jun'ichi Semba ◽

Noriko Tanaka ◽

Maki Wakuta ◽

Tetsuya Suhara

Keyword(s):

Mrna Expression ◽

Mesolimbic Dopamine ◽

Adult Rats ◽

The Brain

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Abstract 17: Early Life Stress Sensitizes The Angiotensin II-elicited Hypertensive Response

Hypertension ◽

10.1161/hyp.76.suppl_1.17 ◽

2020 ◽

Vol 76 (Suppl_1) ◽

Author(s):

Baojian Xue ◽

Terry Beltz ◽

Fang Guo ◽

David M Pollock ◽

Jennifer S Pollock ◽

...

Keyword(s):

Mrna Expression ◽

Proinflammatory Cytokines ◽

Life Stress ◽

Early Life ◽

Early Life Stress ◽

Male Rats ◽

Sprague Dawley ◽

Cns Inflammation ◽

Wide Range ◽

The Brain

Separation of neonatal rodent pups from their mothers has been used as a model to study the effects of early life stress (ELS) on behavioral and physiological responses in adults. Using an Induction-Delay-Expression experimental paradigm, our previous studies demonstrate that a wide range of stressors administered during an induction period produces hypertensive response sensitization (HTRS) in response to a subsequent pro-hypertensive stimulus. HTRS is accompanied by activation of the brain renin-angiotensin system (RAS) and CNS inflammation. The present study investigated whether ELS induces HTRS and changes in brain-related underlying mechanisms. Rat neonates from Sprague-Dawley breeders were subjected to ELS by separating them each morning from their mothers for 3 h on postnatal days 2 to 14. Pups from non-handled litters formed control groups. At 10 weeks of age, male rats were used to evaluate blood pressure and autonomic function using telemetric probes and pharmacological methods. In addition, in separate control and ELS groups, the lamina terminalis (LT) structures and the hypothalamic paraventricular nucleus (PVN) were analyzed for mRNA expression of RAS components and proinflammatory cytokines. Adult ELS rats as compared to non-separated controls exhibited 1) HTRS during expression testing using 2 week ANG II infusions (120 ng/kg/min s.c.; ELS animals, Δ45.5±4.5 mmHg vs. controls, Δ22.4±3.1 mmHg); 2) a greater reduction in mean arterial pressure following ganglionic blockade (hexamethonium, 30 mg/kg, ip), 3) increased sympathetic drive to the heart (atenolol, 8 mg/kg, ip), 4) decreased vagal tone (atropine, 8 mg/kg, ip), and 5) increased mRNA expression of several components of the brain RAS and proinflammatory cytokines in the LT and PVN. These results suggest that maternal ELS may predispose individuals to hypertension that is mediated by upregulation of the brain RAS and proinflammatory cytokines and increased sympathetic drive to the cardiovascular system.

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Nora Virus VP4b and ORF1 Circulate in Hemolymph of Infected D. melanogaster with Coordinate Expression of Vago and Vir-1

Vaccines ◽

10.3390/vaccines8030491 ◽

2020 ◽

Vol 8 (3) ◽

pp. 491

Author(s):

Amanda Macke ◽

Wilfredo Lopez ◽

Darby J. Carlson ◽

Kimberly A. Carlson

Keyword(s):

Rna Virus ◽

Primary Site ◽

The Novel ◽

Qrt Pcr ◽

Basal Expression ◽

Virus Rna ◽

Nora Virus ◽

Candidate Target ◽

The Brain ◽

Insight Into

Study of the novel RNA virus, Nora virus, which is a persistent, picorna-like virus that replicates in the gut of Drosophila melanogaster offers insight into human innate immunity and other picorna-like viruses. Nora virus infection leads to a locomotor abnormality and upregulation of two candidate target proteins, Vago and Virus-induced RNA 1 (Vir-1). These proteins are uncharacterized in response to Nora virus. We hypothesize that Nora virus is circulating in the hemolymph of Nora virus-infected D. melanogaster, allowing for migration beyond the primary site of replication in the gut. Analysis by qRT-PCR demonstrated biphasic viral load and corresponding vago and vir-1 transcription levels, suggesting transcription of vago and vir-1 occurs in response to viral infection. However, Vir-1 is also present in virus-free D. melanogaster suggesting basal expression or alternative functions. Presence of Nora virus RNA and the Viral Protein 4b (VP4b), in hemolymph of infected D. melanogaster supports the hypothesized circulation of Nora virus in the hemolymph. The study suggests that impaired locomotor function may be due to transport of Nora virus from the gut to the brain via the hemolymph.

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Anatomical Distribution and Cellular Basis for High Levels of Aromatase Activity in the Brain of Teleost Fish: Aromatase Enzyme and mRNA Expression Identify Glia as Source

Journal of Neuroscience ◽

10.1523/jneurosci.21-22-08943.2001 ◽

2001 ◽

Vol 21 (22) ◽

pp. 8943-8955 ◽

Cited By ~ 228

Author(s):

Paul M. Forlano ◽

David L. Deitcher ◽

Dean A. Myers ◽

Andrew H. Bass

Keyword(s):

Mrna Expression ◽

Teleost Fish ◽

Aromatase Activity ◽

Anatomical Distribution ◽

Cellular Basis ◽

The Brain

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Molecular cloning and differential expression of three GnRH mRNAs in discrete brain areas and lymphocytes in red drum

Journal of Endocrinology ◽

10.1677/joe.1.06423 ◽

2006 ◽

Vol 188 (3) ◽

pp. 407-416 ◽

Cited By ~ 17

Author(s):

J Shaik Mohamed ◽

I A Khan

Keyword(s):

Mrna Expression ◽

Sea Bass ◽

Red Drum ◽

Sciaenops Ocellatus ◽

Sea Bream ◽

Hypothalamic Area ◽

Cdna Sequences ◽

Peptide Expression ◽

Gonadal Recrudescence ◽

The Brain

Three gonadotropin releasing hormones, seabream GnRH (GnRH-I), chicken GnRH-II (GnRH-II) and salmon GnRH (GnRH-III) cDNAs were isolated from the brain of the red drum, Sciaenops ocellatus. The GnRH cDNA sequences of red drum showed more similarity to those of Atlantic croaker, sea bass and sea bream. The real-time quantitative RT-PCR study revealed expression of GnRH-I and GnRH III mRNAs in the olfactory bulb plus telencephalon (OB+TEL), and preoptic-anterior hypothalamic area (POAH), indicating an overlap of the GnRH-I and GnRH-III systems in these forebrain regions. However, GnRH-II mRNA expression was detected only in the midbrain tegmentum (MT). The GnRH-I mRNA expression in the POAH in fish undergoing gonadal recrudescence was significantly higher than that in gonadally immature individuals, suggesting involvement of the POAH GnRH-I in gonadal maturation. On the other hand, GnRH-III mRNA expression was significantly higher in the OB+TEL region compared with that in the POAH. Moreover, the demonstration of GnRH-I mRNA and peptide expression in red drum lymphocytes indicates that the GnRH-I is synthesized in these cells of the immune system similar to the situation in mammals.

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Analysis of the Expression and Polymorphism ofAPOE, HSP, BDNF,andGRIN2BGenes Associated with the Neurodegeneration Process in the Pathogenesis of Primary Open Angle Glaucoma

BioMed Research International ◽

10.1155/2015/258281 ◽

2015 ◽

Vol 2015 ◽

pp. 1-14 ◽

Cited By ~ 10

Author(s):

Alicja Nowak ◽

Ireneusz Majsterek ◽

Karolina Przybyłowska-Sygut ◽

Dariusz Pytel ◽

Katarzyna Szymanek ◽

...

Keyword(s):

Mrna Expression ◽

Nerve Fiber ◽

Primary Open Angle Glaucoma ◽

Open Angle Glaucoma ◽

Blood Analysis ◽

Specific Gene ◽

Qrt Pcr ◽

Pcr Rflp ◽

Allele Specific ◽

Peripheral Blood Analysis

Glaucoma is characterized by optic neuropathy of the RGC or retinal nerve fiber. The aim of this study was to evaluate a relationship between the neurodegenerative genes’ polymorphisms of theAPOE(rs449647),BDNF(rs2030324),GRIN2B(rs3764028), andHSP70-1(rs1043618) and the occurrence risk of POAG and to investigate its effect on allele-specific gene expression. Genomic DNA was extracted from peripheral blood. Analysis of the genes’ polymorphisms was performed using PCR-RFLP. The level of mRNA expression was determined by QRT-PCR. We showed a statistically significant association ofBDNFandAPOEgenes’ polymorphisms with a risk of POAG occurrence. There was a statistically significant association of the rs2030324 polymorphism with progression of POAG based on cup disc ratio value and rs1043618 polymorphism based on nerve fiber index and rim area. Furthermore, we found that meanHSP70-1mRNA expression was significantly lower in the case of individuals with the G/G genotype than in the case of minor allele carriers, that is, G/C and C/C. We also found thatBDNFandHSP70-1expression level are associated with the progression of POAG based on rim area value. In conclusion, our results suggest thatBDNF,APOE, andHSP70-1genes might be associated with a risk of POAG occurrence in the Polish population.

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Mucin-1 protein and mRNA expression in breast cancer: Predictive value for therapy response and survival after neoadjuvant chemotherapy.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.522 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. 522-522

Author(s):

Bruno Valentin Sinn ◽

Gunter Von Minckwitz ◽

Carsten Denkert ◽

Holger Eidtmann ◽

Silvia Darb-Esfahani ◽

...

Keyword(s):

Breast Cancer ◽

Mrna Expression ◽

Predictive Value ◽

Pathologic Complete Response ◽

Therapy Response ◽

Lower Probability ◽

Muc1 Expression ◽

Mrna Levels ◽

Mucin 1 ◽

Qrt Pcr

522 Background: Mucin-1 (MUC1) is expressed in normal epithelial cells and in breast cancer. Immunotherapeutic agents targeting MUC1 are currently tested in clinical trials. Methods: We evaluated the frequency of MUC1 expression by immunohistochemistry (IHC; n = 691) and quantitative RT-PCR (qRT-PCR; n = 286) in formalin-fixed paraffin-embedded (FFPE) pre-treatment biopsies from patients of the GeparTrio trial (NCT 00544765). mRNA levels were analyzed as a continuous variable, a distribution-based cut-off was chosen for IHC data. The predictive value for therapy response and survival after neoadjuvant anthracycline/taxane-based chemotherapy (CTX) was evaluated. Results: MUC1 was detectable by IHC in 95%. qRT-PCR covered a dynamic range of 3 orders of magnitude. IHC and mRNA data were correlated (p < 0.001). MUC1 was detected in higher quantities in HR+ tumors (p < 0.001), the lowest levels were found in HR-/HER2- tumors (p < 0.001). High MUC1 protein and mRNA expression were associated with lower probability of pathologic complete response (pCR) in the overall population (p < 0.001) and in HR+ (p = 0.004 and < 0.001), HER2- (p < 0.001) and HR+/HER2- tumors (p < 0.001). In multivariable logistic regression, MUC1 was independently predictive (high MUC1 protein: odds ratio (OR) 0.464, 95% CI 0.300 – 0.719, p = 0.001; MUC1 mRNA (per 20-dCT unit): OR 0.673, CI 0.546 – 0.829, p < 0.001). MUC1 protein and mRNA expression were associated with longer patient survival in the overall population (p = 0.03 and < 0.001) and in HER2- tumors (p = 0.005 and < 0.001). MUC1 mRNA was also prognostic in HR+ (p < 0.001) and HR+/HER2- tumors (p = 0.001). In multivariable analysis, protein and mRNA expression were independently prognostic (high MUC1 protein: hazard ratio (HR) 0.388, CI 0.166 – 0.907, p = 0.029; MUC1 mRNA per 20-dCT unit: HR 0.763, CI 0.595 – 0.909, p = 0.005). Conclusions: MUC1 is frequently expressed in breast cancer and detectable by IHC and qRT-PCR from FFPE tissue. Despite its association with HR+ status, it provides independent predictive information for therapy response and survival after neoadjuvant CTX. In clinical immunotherapy trials, MUC1 expression may serve as a predictive marker.

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