chitinase mrna levels by quantitative pcr using the single standard dna acidic mammalian chitinase is a major transcript in the mouse stomach几丁质酶mrna水平定量pcr使用单一标准dna酸性哺乳动物几丁质酶是一个主要记录鼠标胃里.pdfVIP

Chitinase mRNA Levels by Quantitative PCR Using the Single Standard DNA: Acidic Mammalian Chitinase Is a Major Transcript in the Mouse Stomach . . Misa Ohno , Kyoko Tsuda , Masayoshi Sakaguchi, Yasusato Sugahara, Fumitaka Oyama* Department of Applied Chemistry, Kogakuin University, Hachioji, Tokyo, Japan Abstract Chitinases hydrolyze the b-1-4 glycosidic bonds of chitin, a major structural component of fungi, crustaceans and insects. Although mammals do not produce chitin or its synthase, they express two active chitinases, chitotriosidase (Chit1) and acidic mammalian chitinase (AMCase). These mammalian chitinases have attracted considerable attention due to their increased expression in individuals with a number of pathological conditions, including Gaucher disease, Alzheimer’s disease and asthma. However, the contribution of these enzymes to the pathophysiology of these diseases remains to be determined. The quantification of the Chit1 and AMCase mRNA levels and the comparison of those levels with the levels of well-known reference genes can generate useful and biomedically relevant information. In the beginning, we established a quantitative real-time PCR system that uses standard DNA produced by ligating the cDNA fragments of the target genes. This system enabled us to quantify and compare the expression levels of the chitinases and the reference genes on the same scale. We found that AMCase mRNA is synthesized at extraordinarily high levels in the mouse stomach. The level of this mRNA in the mouse stomach was 7- to 10-fold higher than the levels of the housekeeping genes and was comparable to that the level of the mRNA for pepsinogen C (progastricsin), a major component of the gastric mucosa. Thus, AMCase mRNA is a major transcript in mouse stomach, suggesting that AMCase functions as a dig