Genes associated with the formation of germ cells from embryonic stem cells in cultures containing different glucose concentrations.
In a previous study, we established a system for visualizing the development of germ cells from mouse embryonic stem (ES) cells in culture using knock-in ES clones in which visual reporter genes were expressed from the mouse vasa homolog, Mvh. While assessing various culture conditions, we found that germ-cell formation was markedly depressed in low glucose medium. Using a repeated polymerase chain reaction (PCR) subtraction method, we identified genes that were differentially expressed in low versus high glucose media. Three genes that were predominantly expressed in high glucose medium, thioredoxin-interacting protein (Txnip), pituitary tumor-transforming gene 1 (Pttg), and RuvB-like protein 2 (RuvBl2), were further investigated. These genes were also found to be highly expressed in adult and embryonic gonads, and RuvBl2 in particular, which encodes an ATP-dependent DNA helicase, was specifically detected in the spermatocytes and spermatids of the adult testis as well as in primordial germ cells. Furthermore, using a green fluorescent protein (GFP) fusion construct, we found that RuvBl2 was expressed in both the nucleus and cytoplasm of testicular germ cells. These findings suggest a possible relationship between glucose metabolism and germ-cell development.
Mizuno
Kentaro
K
Mitsubishi Kagaku Institute of Life Sciences, Minami-Ooya, Machida, Tokyo, Japan.
Tokumasu
Ako
A
Nakamura
Aya
A
Hayashi
Yutaro
Y
Kojima
Yoshiyuki
Y
Kohri
Kenjiro
K
Noce
Toshiaki
T
D000074183:ATPases Associated with Diverse Cellular Activities; D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D053487:DEAD-box RNA Helicases / Q000096:biosynthesis / Q000235:genetics; D004265:DNA Helicases / Q000096:biosynthesis / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D005947:Glucose / Q000378:metabolism* / Q000502:physiology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D010063:Ovum / Q000166:cytology / Q000378:metabolism*; D064249:Securin; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*; D013737:Testis / Q000378:metabolism; D013879:Thioredoxins / Q000096:biosynthesis / Q000235:genetics
D002352:Carrier Proteins; D009363:Neoplasm Proteins; D064249:Securin; C445672:Txnip protein, mouse; D013879:Thioredoxins; C090864:Ddx4 protein, mouse; D000074183:ATPases Associated with Diverse Cellular Activities; D004265:DNA Helicases; C518330:RUVBL2 protein, mouse; D053487:DEAD-box RNA Helicases; D005947:Glucose