Detection and assessment of androgenic potency of endocrine-disrupting chemicals using three-spined stickleback, Gasterosteus aculeatus.
The male three-spined stickleback (Gasterosteus aculeatus) produces a glue protein named "spiggin" that is used as a cementing substance for building its nest. The synthesis of spiggin is under the control of androgenic stimulation. Therefore, spiggin is an effective biomarker of any androgenic activity displayed by environmental chemicals, similarly to the use of vitellogenin as an estrogenic biomarker. The aim of this study was to establish a quantification system for spiggin mRNA to develop a highly sensitive system for evaluating environmental androgens. In this process, two different types of cDNA encoding spiggin (SPG-1 and SPG-2) were isolated. They closely resemble each other in primary structure and features. In addition, the transcriptions of both spiggin gene were induced by only androgenic stimulation in a receptor-mediated manner. These findings suggest the multiplicity albeit specificity of spiggin in the stickleback. The quantification system for spiggin mRNA was established using a real-time RT-PCR technique. This system enables accurate quantification within a wide range of spiggin mRNA from 10(1) to 10(6) copies.
Nagae
Masaki
M
Faculty of Environmental Studies, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, Nagasaki 852-8521, Japan. nagae@net.nagasaki-u.ac.jp
Kawasaki
Fumie
F
Tanaka
Yuka
Y
Ohkubo
Nobuyuki
N
Matsubara
Takahiro
T
Soyano
Kiyoshi
K
Hara
Akihiko
A
Arizono
Koji
K
Scott
Alexander P
AP
Katsiadaki
Ioanna
I
D000595:Amino Acid Sequence; D000728:Androgens / Q000819:agonists / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D052244:Endocrine Disruptors / Q000032:analysis* / Q000633:toxicity*; D005260:Female; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D007668:Kidney; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012015:Reference Standards; D017386:Sequence Homology, Amino Acid; D012727:Sex Characteristics; D023701:Smegmamorpha / Q000235:genetics / Q000378:metabolism*
D000728:Androgens; D018076:DNA, Complementary; D052244:Endocrine Disruptors; D029941:Fish Proteins; D012333:RNA, Messenger