Laboratory diagnosis of enteroviral infections of the central nervous system by using a nested RT-polymerase chain reaction (PCR) assay

2003 ◽  
Vol 47 (4) ◽  
pp. 557-562 ◽  
Author(s):  
Cakir Guney ◽  
Etem Ozkaya ◽  
Mehmet Yapar ◽  
Ismail Gumus ◽  
Ayhan Kubar ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Laboratory Diagnosis ◽  
Chain Reaction ◽  
Pcr Assay ◽  
The Central Nervous System ◽  
Polymerase Chain

Expression of the integrin alpha 6 beta 4 in peripheral nerves: localization in Schwann and perineural cells and different variants of the beta 4 subunit

1994 ◽  
Vol 107 (2) ◽  
pp. 543-552 ◽  
Author(s):  
C.M. Niessen ◽  
O. Cremona ◽  
H. Daams ◽  
S. Ferraresi ◽  
A. Sonnenberg ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Peripheral Nerves ◽  
Northern Blot Analysis ◽  
The Other ◽  
Chain Reaction ◽  
The Central Nervous System ◽  
Polymerase Chain ◽  
Integrin Alpha 6

Integrin alpha 6 beta 4 is expressed in human peripheral nerves, but not in the central nervous system. This integrin heterodimer has previously been found in perineural fibroblast-like cells and in Schwann cells (SCs), which both assemble a basement membrane but do not form hemidesmosomes. We show here that in SCs, which had formed a myelin sheath, alpha 6 beta 4 was enriched in the proximity of the nucleus, at Ranvier paranodal areas and at Schmitt-Lanterman clefts; alpha 6 beta 4 was also found at the grooved interface between small axons and non-myelinating SCs. Immunoprecipitation of human peripheral nerves, in combination with Western blotting showed that beta 4 is associated with the alpha 6A subunit. Northern blot analysis of human peripheral nerves showed a single beta 4 transcript of 6 kb. Using the reverse transcriptase polymerase chain reaction, we detected two mRNA species, one for the most common (−70, -53) form of beta 4 and the other encoding the (+53) variant of beta 4. Cultured SCs were devoid of alpha 6 beta 4 but expressed alpha 6 beta 1, indicating that SCs lose beta 4 expression when contact with neurons is lost. Thus, resting SCs in contact with axons express alpha 6A in combination with beta 4, irrespective of myelin formation. We suggest that alpha 6 beta 4 expressed in SCs plays a role in peripheral neurogenesis.

scholarly journals Quantitative RT-PCR Analysis of Uncoupling Protein Isoforms in Mouse Brain Cortex: Methodological Optimization and Comparison of Expression with Brown Adipose Tissue and Skeletal Muscle

2004 ◽  
Vol 24 (7) ◽  
pp. 780-788 ◽  
Author(s):  
Sylvain Lengacher ◽  
Pierre J. Magistretti ◽  
Luc Pellerin
Keyword(s):  
Skeletal Muscle ◽  
Central Nervous System ◽  
Polymerase Chain Reaction ◽  
Nervous System ◽  
Adipose Tissue ◽  
Brain Cortex ◽  
Chain Reaction ◽  
Brown Adipose ◽  
The Central Nervous System ◽  
Polymerase Chain

Uncoupling proteins (UCPs) present in the inner mitochondrial membrane are involved in uncoupling respiration from ATP synthesis. Five UCP isoforms have been identified but information about their presence and level of expression in the central nervous system remains incomplete. To determine the nature and proportion of UCP isoform mRNAs present in brain cortex, we developed and optimized a specific quantitative reverse-transcription polymerase chain reaction procedure. Optimal range of RNA concentrations to be used in the reverse-transcriptase reaction was determined. Primer design and concentration were optimized for each target gene while polymerase chain reaction efficiency was assessed for a range of reverse-transcriptase dilutions. Genomic contribution to the quantitative signal was evaluated for each isoform and minimized. Three reference genes were tested for normalization, and β-actin was found to be the most stable among tissues. Results indicate that brain cortex contains significant amounts of all UCP mRNAs, with UCP5 and UCP4 being the most abundant, as opposed to brown adipose tissue and skeletal muscle, which predominantly express UCP1 and UCP3, respectively. These data provide a first quantitative assessment of UCP mRNA expression in mouse brain, showing the presence of all five isoforms with distinct proportions, thus suggesting specific roles in the central nervous system.

Sign in / Sign up

Export Citation Format

Share Document