Protocol for RNA isolation and RT‐PCR confirmation for Neomycin gene expression in transformed Bodo saltans v1

protocols.io ◽  
2021 ◽  
Author(s):  
Fatma Gomaa ◽  
Zhu-Hong Li ◽  
Roberto Docampo ◽  
Virginia Edgcomb
Keyword(s):  
Gene Expression ◽  
Rna Isolation ◽  
Rt Pcr

Comparative Expression Analysis of Genes Encoding Neurotransmitter Receptors Between Entrepreneurs and Non-Entrepreneurs

2021 ◽  
Vol 15 (9) ◽  
pp. 3058-3062
Author(s):  
Fatih Yildirim ◽  
M. Enes Aslan ◽  
Hüseyin Daştan ◽  
M. Emirhan Kula ◽  
Seda Kayapaliyildirim ◽  
...  
Keyword(s):  
Gene Expression ◽  
High Capacity ◽  
Rna Isolation ◽  
Neurotransmitter Receptors ◽  
Pcr Array ◽  
Rt Pcr ◽  
Gene Expressions ◽  
Blood Samples ◽  
Genes Encoding ◽  
Neurotransmitter Activity

Introduction and Purpose: Recent studies revealed that different behavioral and physiological processes are closely related to different receptor levels in humans. On the other hand, there weren’t any studies that investigated neurotransmitter activity and entrepreneurship relationships. Little is known about how genes expressed in the blood are associated with entrepreneurship. In this study, we measured the expression of 84 genes encoding neurotransmitter receptors between the entrepreneurs and non-entrepreneurs (n=25) using RT-PCR arrays to monitor differentially expressed genes for exploring molecular mechanism behind the entrepreneurship mindset. Materials and Methods: After determining whether participants are entrepreneurs or not, blood samples were collected. Blood samples were collected in Vacutainer® EDTA tubes as 10 ml and RNA isolation was performed at the Erzurum Technical University, Cell Culture Laboratory. Trizol® solution for RNA isolation (Thermo®, USA) was applied according to the manufacturer's instructions. Concentrations of RNA samples were measured at 260 nm using a spectrophotometer (Epoch®, Biotek). Then, total RNA was reverse transcribed into cDNA by using High-Capacity cDNA Reverse Transcription Kit (Thermo®). Total 84 genes were analyzed via RT² Profiler™ Human Neurotransmitter Receptors PCR Array (Qiagen®, USA) in Rotor-Gene Q real-time PCR cycler (Qiagen®, USA). Findings: The gene expression results obtained from RT-PCR were compared between entrepreneurs and non-entrepreneurs, and presented as Fold Changes (FC). According to our results, positive FC values indicated an increase in the expression of the genes and negative FC values indicated decrease in gene expression levels in entrepreneurs. Results: These 84 different genes regulate neurotransmitter biosynthesis, uptake, transport, and signaling via neurotransmitter receptors. According to gene expression analyses, gene expressions that could be related to the entrepreneur behavior might be connected to not only undesired psychological outcomes like various addictions and also neurological cases such as schizophrenia and depressive disorder. Our results firstly indicated that entrepreneurship was not only associated with neurotransmitter release but also with receptor levels. Keywords: Entrepreneurship, Entrepreneurs, Genes, Neurotransmitter Receptors, RT-PCR Array

Comparative Expression Analysis of Genes Encoding Neurotransmitter Receptors Between Entrepreneurs and Non-Entrepreneurs

2021 ◽  
Vol 15 (10) ◽  
pp. 2928-2932
Author(s):  
Fatih Yildirim ◽  
M. Enes Aslan ◽  
Hüseyin daştan ◽  
M. Emirhan Kula ◽  
Seda Kayapali Yildirim ◽  
...  
Keyword(s):  
Gene Expression ◽  
High Capacity ◽  
Rna Isolation ◽  
Neurotransmitter Receptors ◽  
Pcr Array ◽  
Rt Pcr ◽  
Gene Expressions ◽  
Blood Samples ◽  
Genes Encoding ◽  
Neurotransmitter Activity

Introduction and Purpose: Recent studies revealed that different behavioral and physiological processes are closely related to different receptor levels in humans. On the other hand, there weren’t any studies that investigated neurotransmitter activity and entrepreneurship relationships. Little is known about how genes expressed in the blood are associated with entrepreneurship. In this study, we measured the expression of 84 genes encoding neurotransmitter receptors between the entrepreneurs and non-entrepreneurs (n=25) using RT-PCR arrays to monitor differentially expressed genes for exploring molecular mechanism behind the entrepreneurship mindset. Materials and Methods: After determining whether participants are entrepreneurs or not, blood samples were collected. Blood samples were collected in Vacutainer® EDTA tubes as 10 ml and RNA isolation was performed at the Erzurum Technical University, Cell Culture Laboratory. Trizol® solution for RNA isolation (Thermo®, USA) was applied according to the manufacturer's instructions. Concentrations of RNA samples were measured at 260 nm using a spectrophotometer (Epoch®, Biotek). Then, total RNA was reverse transcribed into cDNA by using High-Capacity cDNA Reverse Transcription Kit (Thermo®). Total 84 genes were analyzed via RT² Profiler™ Human Neurotransmitter Receptors PCR Array (Qiagen®, USA) in Rotor-Gene Q real-time PCR cycler (Qiagen®, USA). Findings: The gene expression results obtained from RT-PCR were compared between entrepreneurs and non-entrepreneurs, and presented as Fold Changes (FC). According to our results, positive FC values indicated an increase in the expression of the genes and negative FC values indicated decrease in gene expression levels in entrepreneurs. Results: These 84 different genes regulate neurotransmitter biosynthesis, uptake, transport, and signaling via neurotransmitter receptors. According to gene expression analyses, gene expressions that could be related to the entrepreneur behavior might be connected to not only undesired psychological outcomes like various addictions and also neurological cases such as schizophrenia and depressive disorder. Our results firstly indicated that entrepreneurship was not only associated with neurotransmitter release but also with receptor levels. Keywords: Entrepreneurship, Entrepreneurs, Genes, Neurotransmitter Receptors, RT-PCR Array

1650: Novel Gene Expression Markers in Renal Tumors Using Rt-PCR on Fixed Tissues Useful in Differentiating Histologic Subtypes

2004 ◽  
Vol 171 (4S) ◽  
pp. 436-436
Author(s):  
John A. Petros ◽  
Audry N. Schuetz ◽  
Andrew N. Young ◽  
Q. Yin Goen ◽  
So Dug Lim ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Tumors ◽  
Rt Pcr ◽  
Novel Gene ◽  
Histologic Subtypes

Gene expression of collagen, fibronectin and elastin on a collagen and a collagen-fabric matrix measured by realtime RT-PCR

2004 ◽  
Vol 52 (S 1) ◽  
Author(s):  
LO Schilling ◽  
N Davies ◽  
R Autschbach ◽  
P Zilla
Keyword(s):  
Gene Expression ◽  
Rt Pcr

The Protective Effect of Metformin against Oxandrolone-Induced Infertility in Male Rats

2020 ◽  
Vol 26 ◽  
Author(s):  
Abdulqader Fadhil Abed ◽  
Yazun Bashir Jarrar ◽  
Hamzeh J Al-Ameer ◽  
Wajdy Al-Awaida ◽  
Su-Jun Lee
Keyword(s):  
Gene Expression ◽  
Male Infertility ◽  
Protective Effect ◽  
Histological Examination ◽  
Sperm Count ◽  
Serum Testosterone ◽  
Male Rats ◽  
P Value ◽  
Protective Effects ◽  
Rt Pcr

Background: Oxandrolone is a synthetic testosterone analogue that is widely used among bodybuilders and athletes. However, oxandrolone causes male infertility. Recently, it was found that metformin reduces the risk of infertility associated with diabetes mellitus. Aim: This study aimed to investigate the protective effects of metformin against oxandrolone-induced infertility in male rats. Methods: Rats continuously received one of four treatments (n=7) over 14 days: control DMSO administration, oxandrolone administration, metformin administration, or co-administration of oxandrolone and metformin. Doses were equivalent to those used for human treatment. Subsequently, testicular and blood samples were collected for morphological, biochemical, and histological examination. In addition, gene expression of the testosterone synthesizing enzyme CYP11A1 was analyzed in the testes using RT-PCR. Results: Oxandrolone administration induced male infertility by significantly reducing relative weights of testes by 48%, sperm count by 82%, and serum testosterone levels by 96% (ANOVA, P value < 0.05). In addition, histological examination determined that oxandrolone caused spermatogenic arrest which was associated with 2-fold downregulation of testicular CYP11A1 gene expression. However, co-administration of metformin with oxandrolone significantly ameliorated toxicological alterations induced by oxandrolone exposure (ANOVA, P value < 0.05). Conclusion: Metformin administration protected against oxandrolone-induced infertility in male rats. Further clinical studies are needed to confirm the protective effect of metformin against oxandrolone-induced infertility among athletes.

Global expression analysis of ECL cells in Mastomys natalensis gastric mucosa identifies alterations in the AP-1 pathway induced by gastrin-mediated transformation

2004 ◽  
Vol 20 (1) ◽  
pp. 131-142 ◽  
Author(s):  
M. Kidd ◽  
T. Hinoue ◽  
G. Eick ◽  
K. D. Lye ◽  
S. M. Mane ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gastric Mucosa ◽  
Western Blot ◽  
Cell Transformation ◽  
Mastomys Natalensis ◽  
Rt Pcr ◽  
Cell Hyperplasia ◽  
Ecl Cells ◽  
Ecl Cell ◽  
Gene Expression Alterations

Enterochromaffin-like (ECL) cell hyperplasia and then irreversible neoplasia can be generated in the African rodent Mastomys natalensis using the H2 receptor blocker, loxtidine, for 8–16 wk. We used a GeneChip approach complemented by standard technologies to identify gene expression alterations in the gastric mucosa during gastrin-mediated ECL cell transformation. Gastric mucosa (mucosal scrapping) and ECL cell-enriched fractions were obtained from untreated Mastomys (controls) and from animals treated with loxtidine for 8 wk (hyperplasia). Tumor ECL cells were obtained by hand-dissection of gastric ECL cell nodules from animals treated with loxtidine for >16 wk and from a spontaneously developed ECL cell tumor. RNA was isolated, examined on rat U34A GeneChips, and comparison analysis was performed to identify altered gene expression. Alterations in gene expressions were examined further by immunohistochemistry, quantitative RT-PCR (Q-RT-PCR), sequencing and Western blot. GeneSpring analysis demonstrated alterations in few genes (<20) in hyperplastic and tumor mucosa. The histamine H1 receptor was consistently increased in proliferating mucosa. This gene change was confirmed by Q-RT-PCR. Other genes showing alterations included neural-(chromogranin A and somatostatin), cell-cycle-, and AP-1-associated genes. Immunostaining confirmed alterations in neural markers. Cluster analysis of ECL cell-enriched samples demonstrated that c- fos and junD were differently regulated. Q-RT-PCR and Western blot in prospectively collected gastric mucosal samples confirmed the differential expression of Fos and Jun. The negative regulators of AP-1, JunD, and Menin were decreased in tumor mucosa. A missense of unknown function was noted in the menin gene. Hypergastrinemia in an animal model of gastric carcinoids differentially altered the histamine type 1 receptor and gene expression and protein composition of AP-1. These results suggest that expression of this receptor and an altered composition of AP-1 with a loss of inhibition play a role in ECL cell transformation.

Cloning of rabbit prolactin cDNA and prolactin gene expression in the rabbit mammary gland

1996 ◽  
Vol 16 (1) ◽  
pp. 27-37 ◽  
Author(s):  
L Gabou ◽  
M Boisnard ◽  
I Gourdou ◽  
H Jammes ◽  
J-P Dulor ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammary Gland ◽  
Amino Acid ◽  
Untranslated Regions ◽  
Pcr Analysis ◽  
Cdna Clones ◽  
Rt Pcr ◽  
Prolactin Gene ◽  
New Zealand White Rabbits ◽  
Rna Probe

ABSTRACT cDNA clones coding for rabbit prolactin were isolated from a pituitary library using a rat prolactin RNA probe. One cDNA contained 873 bases including the entire coding sequence of rabbit prolactin, its signal peptide and the 5′ and 3′ untranslated regions of 44 and 145 nucleotides respectively. The deduced amino acid sequence of the cloned prolactin cDNA presented a 93–78% identity with mink, porcine and human prolactins. The prolactin gene transcription was investigated by RT-PCR analysis in several organs of midlactating New Zealand White rabbits. The ectopic transcription of the prolactin gene was examined in more detail in the mammary gland. A strong PCR signal was detected in the mammary gland of virgin does and was also observed during pregnancy and at the beginning of lactation. This PCR signal was very weak in mid-lactating and absent in post-weaning mammary gland.

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