Biochemical and Biophysical Research Communications
A novel marker for terminal Schwann cells, homocysteine-responsive ER-resident protein, as isolated by a single cell PCR-differential display
Section snippets
Materials and methods
Animals. Adult and developing Wistar rats (12-week-old female and 14-day-old male) were purchased from a breeder (SLC, Japan), 1 week prior to experiments, and they were maintained in a temperature- and humidity-controlled animal facility with a 12:12-h light:dark cycle. All the animals were treated and/or handled in accordance with the “Recommendations for Handling of Laboratory Animals for Biomedical Research,” compiled by the Committee on the Safety and Ethical Handling Regulations for
Screening of genes marking TSCs
To identify genes specifically expressed in TSCs, we performed a PCR-based DD screening of two types of Schwann cells, viz., TSCs and MSCs. Herein, we applied a method of uni-directional synthesis and amplification of enriched full-length cDNA to RNA purified from a single cell, and we could obtain total cDNA fraction as moderately strong smears ranging from 0.2 to 4.0 kb with scattering several bright bands. Representative data are shown in Fig. 1A.
To check the quality and the origin of the
Discussion
We have identified a gene called Herp, as a novel molecular marker for terminal Schwann cells, by applying a single cell PCR-DD.
Myelinating Schwann cells (MSCs) are known to contribute to axonal outgrowth and guidance in the regeneration process after nerve injury. On the other hand, terminal Schwann cells (TSCs) appear to play critical roles in maintaining the structure and function of neuromuscular junctions (NMJs) as well as in the repair process. Whereas molecular characteristics of MSCs
Acknowledgements
This work was supported in part by Grant-in-Aid for Scientific Research (B) (to S.F.) and Grant-in-Aid for Young Scientists (A) (to T.Y.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by research grants from National Cardiovascular Center of Japan (to S.F.).
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