Dysfunction of Ca2+/ CaM kinase IIA cascades in basolateral amygdala of posttraumatic stress disorder rats

2011 ◽  
Vol 26 (S2) ◽  
pp. 178-178
Author(s):  
Y. Shi ◽  
F. Han
Keyword(s):  
Normal Level ◽  
Calcium Level ◽  
Basolateral Amygdala ◽  
Intracellular Free Calcium ◽  
Free Calcium ◽  
Rt Pcr ◽  
Control Groups ◽  
Male Wistar Rats ◽  
Polymerase Chain ◽  
Set Up

ObjectiveTo explore changes of Ca2+-CaM-CaMKIIα in basolateral amygdala of PTSD rats may reveal part of the pathogensis.MethodsThe SPS-method was used to set up the rat PTSD models. A total of 90 male Wistar rats were randomly divided into1d, 4d, 7d, 14d groups of SPS and normal control groups. The intracellular free calcium level in basolateral amygdala was examined by fluorescence spectrophotometer. CaM and CaMKIIα expression in basolateral amygdala were examined by immunohistochemistry, western blotting and reverse transcription-polymerase chain reaction (RT-PCR).ResultsThe intracellular free calcium level reached the peak 1 day after SPS stimulation, then gradually decreased to normal level. The expression of CaM 1day after SPS is also the most and then decreased to normal level. In contrast, CaMKIIα expression showed a significant down-regulation 1day after SPS throughout and then gradually increased to normal level. This findings suggest dysfunction of Ca2+-CaM-CaMKIIα in basolateral amygdala of PTSD rats.ConclusionThus, the trauma-induced enhanced anxiety appear to be associated with, and possibly caused by, changes of Ca2+-CaM-CaMKIIα in basolateral amygdala.

The ratio of macrophage prostaglandin and leukotriene synthesis is determined by the intracellular free calcium level

1990 ◽  
Vol 39 (8) ◽  
pp. 1313-1319 ◽  
Author(s):  
Volkhard Kaever ◽  
Hans-Jürgen Pfannkuche ◽  
Klaus Wessel ◽  
Klaus Resch
Keyword(s):  
Calcium Level ◽  
Intracellular Free Calcium ◽  
Free Calcium

Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Screening of 29 Translocations and Chromosomal Aberrations in Acute Leukemia

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 574-588 ◽  
Author(s):  
Niels Pallisgaard ◽  
Peter Hokland ◽  
Dorthe C. Riishøj ◽  
Bent Pedersen ◽  
Poul Jørgensen
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Reverse Transcription ◽  
Cytogenetic Analysis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Lymphoid Leukemia ◽  
Polymerase Chain ◽  
Set Up

Abstract We have developed a multiplex reverse transcription-polymerase chain reaction (RT-PCR) reaction, which enables us to detect 29 translocations/chromosomal aberrations in patients with acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML). Through the construction and optimization of specific primers for each translocation, we have been able to reduce the set-up to 8 parallel multiplex PCR reactions, thus greatly decreasing the amount of work and reagents. We show the value of our set-up in a retrospective analysis on cryopreserved material from 102 AML and 62 ALL patients. The multiplex RT-PCR detected a hybrid mRNA resulting from a structural chromosomal aberration in 45 of 102 (44%) of the AML and in 28 of 62 (45%) of the pediatric ALL cases. Importantly, in 33% of AML and in 47% of the ALL cases with cytogenetic data, submicroscopic chromosomal aberrations or masked translocations were shown that were not detected in the cytogenetic analysis either for structural reasons or because of an insufficient number of metaphases obtained. This multiplex RT-PCR system, which can handle up to 10 patients with a response time of 2 working days, is thus an important tool that complements cytogenetic analysis in the up-front screening of acute leukemia patients and should provide a rapid and efficient characterization of leukemia cells, even in situations with sparse patient material.

scholarly journals Functional and Molecular Characterization of a Muscarinic Receptor Type and Evidence for Expression of Choline-Acetyltransferase and Vesicular Acetylcholine Transporter in Human Granulosa-Luteal Cells1

1999 ◽  
Vol 84 (5) ◽  
pp. 1744-1750 ◽  
Author(s):  
S. Fritz ◽  
K. J. Föhr ◽  
S. Boddien ◽  
U. Berg ◽  
C. Brucker ◽  
...  
Keyword(s):  
Western Blotting ◽  
Choline Acetyltransferase ◽  
Nerve Fibers ◽  
Intracellular Free Calcium ◽  
Vesicular Acetylcholine Transporter ◽  
Free Calcium ◽  
Rt Pcr ◽  
Natural Ligand ◽  
M1 Receptor

Previously, we provided evidence for the presence of a class of muscarinic receptors on human luteinized granulosa cells (human GC) that is linked to transient increases in intracellular free calcium levels, but not to steroid production. The precise nature of the receptor is not known, and neither its function nor the source of its natural ligand acetylcholine (ACh) is clear. To address these issues we used RT-PCR approaches and isolated complementary DNAs corresponding to the M1 receptor subtype from reverse transcribed human GC messenger ribonucleic acids. M1 receptors were further shown by immunocytochemistry, using a M1 receptor antiserum. Single cell calcium measurements showed that the M1 receptor was functionally active and linked to acute increases in intracellular free calcium, as the M1 receptor specific antagonist pirenzepine blocked the Ca2+-mobilizing effect of oxotremorine M (a muscarinic agonist). An unexpected consequence of M1 receptor activation was evidenced by the ability of muscarinic agonists to stimulate the proliferation of human GC within 24 h. In vivo, ACh, the natural ligand of these receptors is thought to be contained in cholinergic nerve fibers innervating the ovary. Surprisingly, the prerequisite for the synthesis of ACh, the enzyme choline-acetyltransferase (ChAT), is also expressed by human GC, as shown by Western blotting and immunocytochemistry. In addition, these cells express another marker for ACh synthesis, namely the gene for the vesicular acetylcholine transporter, as evidenced by RT-PCR cloning, Western blotting, and immunocytochemistry. In conclusion, our data identify the M1 receptor in human GC and point to a novel, trophic role of the neurotransmitter ACh. Furthermore, the presence of the prerequisites of ACh synthesis in human GC indicate that an autocrine/paracrine regulatory loop also exists in the in vivo counterparts of these cells in the ovary, i.e. in the cells of the preovulatory follicle and/or of the young corpus luteum.

scholarly journals Intracellular free calcium level and its response to cAMP stimulation in developingDictyosteliumcells transformed with jellyfish apoaequorin cDNA

FEBS Letters ◽  
1994 ◽  
Vol 337 (1) ◽  
pp. 43-47 ◽  
Author(s):  
Shweta Saran ◽  
Hajime Nakao ◽  
Masao Tasaka ◽  
Hidetoshi Iida ◽  
Frederick I. Tsuji ◽  
...  
Keyword(s):  
Calcium Level ◽  
Intracellular Free Calcium ◽  
Free Calcium ◽  
Camp Stimulation

scholarly journals A Pipeline with Multiplex Reverse Transcription Polymerase Chain Reaction and Microarray for Screening of Chromosomal Translocations in Leukemia

2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Fei-Fei Xiong ◽  
Ben-Shang Li ◽  
Chun-Xiu Zhang ◽  
Hui Xiong ◽  
Shu-Hong Shen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Fusion Gene ◽  
Chromosomal Rearrangements ◽  
Pcr Primers ◽  
Fusion Genes ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Set Up

Chromosome rearrangements and fusion genes present major portion of leukemogenesis and contribute to leukemic subtypes. It is practical and helpful to detect the fusion genes in clinic diagnosis of leukemia. Present application of reverse transcription polymerase chain reaction (RT-PCR) method to detect the fusion gene transcripts is effective, but time- and labor-consuming. To set up a simple and rapid system, we established a method that combined multiplex RT-PCR and microarray. We selected 15 clinically most frequently observed chromosomal rearrangements generating more than 50 fusion gene variants. Chimeric reverse primers and chimeric PCR primers containing both gene-specific and universal sequences were applied in the procedure of multiplex RT-PCR, and then the PCR products hybridized with a designed microarray. With this approach, among 200 clinic samples, 63 samples were detected to have gene rearrangements. All the detected fusion genes positive and negative were validated with RT-PCR and Sanger sequencing. Our data suggested that the RT-PCR-microarray pipeline could screen 15 partner gene pairs simultaneously at the same accuracy of the fusion gene detection with regular RT-PCR. The pipeline showed effectiveness in multiple fusion genes screening in clinic samples.

Multiplex Reverse Transcription-Polymerase Chain Reaction for Simultaneous Screening of 29 Translocations and Chromosomal Aberrations in Acute Leukemia

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 574-588 ◽  
Author(s):  
Niels Pallisgaard ◽  
Peter Hokland ◽  
Dorthe C. Riishøj ◽  
Bent Pedersen ◽  
Poul Jørgensen
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Leukemia ◽  
Chromosomal Aberrations ◽  
Reverse Transcription ◽  
Cytogenetic Analysis ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Lymphoid Leukemia ◽  
Polymerase Chain ◽  
Set Up

We have developed a multiplex reverse transcription-polymerase chain reaction (RT-PCR) reaction, which enables us to detect 29 translocations/chromosomal aberrations in patients with acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML). Through the construction and optimization of specific primers for each translocation, we have been able to reduce the set-up to 8 parallel multiplex PCR reactions, thus greatly decreasing the amount of work and reagents. We show the value of our set-up in a retrospective analysis on cryopreserved material from 102 AML and 62 ALL patients. The multiplex RT-PCR detected a hybrid mRNA resulting from a structural chromosomal aberration in 45 of 102 (44%) of the AML and in 28 of 62 (45%) of the pediatric ALL cases. Importantly, in 33% of AML and in 47% of the ALL cases with cytogenetic data, submicroscopic chromosomal aberrations or masked translocations were shown that were not detected in the cytogenetic analysis either for structural reasons or because of an insufficient number of metaphases obtained. This multiplex RT-PCR system, which can handle up to 10 patients with a response time of 2 working days, is thus an important tool that complements cytogenetic analysis in the up-front screening of acute leukemia patients and should provide a rapid and efficient characterization of leukemia cells, even in situations with sparse patient material.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain
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