The glycoprotein hormone family consists of luteinizing hormone (LH), follicle stimulating hormone (FSH) and thyroid stimulating hormone, which are secreted by the pituitary gland in all mammalian species, and choriogonadotropin (CG), which is secreted by the placenta in primates and equids. The hormones are composed of a common α subunit and a hormone specific β subunit which are non-covalently associated. Recent advances in biotechnology, particularly in the production of recombinant proteins, have provided opportunities to produce sufficient quantities of recombinant fish GTHs using various expression hosts. Japanese eel Anguilla japonica is one of the most important fish species being aquacultured in Japan but is hampered from the fact that this species does not reproduce in captivity. Artificial induction of gonadal maturation has been successful by administration of pituitary extracts or human chorionic gonadotropin, but the understanding the regulatory mechanism of gonadal development moderated by follicle stimulating hormone (FSH) and luteinizing hormone (LH) remains elusive due to lack of suitable amounts of eel gonadotropins (GTHs). In the present study, we produced tethered rec-eel LH and deglycosylated mutants (56, 79 and 56-79 of α subunit; 10 of β-subunit) of Asn-linked oligosaccharides in CHO suspension cells. Luteinizing hormone acts through binding to its specific receptor. Binding of ligand to the receptor activates the adenosine 3',5'-cyclic monophosphate (cAMP) pathway (McFarland et al., 1989; Ji and Ji, 1991a; Rose, 1998) and the inositol 1 phosphate (IP1) secondary messenger systems. After stimulation of eelLH/CG receptor transfected CHO cells with rec-LH wild type (wt) and mutant hormones as a ligand, production of cAMP and IP-1 were evaluated (Cisbio). cAMP IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56_79 and βΔ10 were 606.2; 374.9; 100.3; 14.2 and 210.9 ng/ml, respectively. IP-1 IC-50 values by rec-eelLH wt; αΔ56; αΔ79; αΔ56.79 and βΔ10 were 28.3; 16.04; 4.3; 2.1 and 3.6ng/ml, respectively too. As seen in both of the second messenger production, general stimulatory pattern is analogous. cAMP and IP-1 stimulation by wild type and αΔ56, as well as αΔ79 and βΔ10 were approximate, but the stimulating effect of double mutant (αΔ56_ 79) was drastically higher. According to the data, deglycosylated eelLH may bind to the receptor with high affinity and cAMP production is gradually increased. Furthermore, receptor activation by tethered rec-eel mutant ligands (FreeStyle CHO-MAX Expression System) will be evaluated with β arrestin recruitment and GPCR internalization for N-linked oligosaccharides’ biological role in activation of eelLH/CGR.

소음 스트레스로 인한 뱀장어(Anguilla japonica)의 영향을 파악하기 위하여 스트레스 지표로 사용되는 코티졸, 포도당, 알부민 및 glucocorticoid receptor(GCR) 유전자의 발현 양을 측정, 분석하여 노출되지 않은 대조구와 비교하였다. 그 결과, 알부민은 노출 1시간 후에 낮은 값을 보인 반면 코티졸과 포도당은 대조구에 비해 매우 큰 차이를 보이며 높게 나타났다. GCR 유전자의 조직 발현 결과 간, 아가미, 근육 및 소장에서 많이 발현하였다. 소음 노출에 따른 시간의 변화에서 간과 아가미 근육과 소장에서 발현이 감소하는 양상을 나타내었다. 실험결과 뱀장어의 glucocorticoid receptor 유전자의 발현변화가 소음 스트레스로 인한 영향을 파악하는데 유용한 지표가 될 수 있음을 확인하였다.

Gene expressions of cytochrome P4501A (CYP1A), aryl hydrocarbon receptor (AhR) and vitellogenin (Vg) by endocrine disruptors, benzo[]pyrene (B[a]P) and tributyltin (TBT) were examined in cultured eel hepatocytes which were isolated from eels treated previously with B[a]P (10 mg/kg) or estradiol- (20 mg/kg) in vivo, and the relationship between CYP1A, AhR and Vg genes were studied. When the cultured eel hepatocytes were treated with B[a]P () the gene expressions of CYP1A and AhR were enhanced in a concentration-dependent manner. However, when treated with TBT () the gene expressions of CYP1A and AhR were suppressed at high concentrations (), while having no effects at low concentrations (). Gene expression of Vg was also suppressed by TBT in a concentration-dependent manner in cultured eel hepatocytes which was previously treated in vivo with estradiol-.

We investigated the effects of temperature changes on the oxygen consumption rhythm in Japanese eels, Anguilla japonica, using an automatic intermittent flow respirometer (AIFR). The endogenous rhythm of the oxygen consumption rate (OCR) in the eels (n = 18; 44-74 cm, 145-690 g), freshly collected by bag net from estuaries, was nearly synchronous with the tidal pattern of the estuarine collection site. The magnitude of mean OCR (mOCR) of eels showed variable range of 82.2 - 116.5 ml O2 kg-1 ww h-1 under constant conditions. In case of increasing temperature from 25 to 38℃, the OCR of eels exhibited a gradually increasing trend with a rhythmic pattern until 36℃. Above 36℃, the rhythms of the OCR dampened and the OCR decreased rapidly at around 36 - 37℃. The OCR of the eels exhibited the maximum value at 38℃, and then it sharply decreased. The results suggested that the critical thermal maximum (CTM) regarding the endogenous rhythms of the eels was at around 36 - 37℃ when water temperature increased at 0.5℃/14 h following the acclimation at 25℃. In case of decreasing temperature (0.5℃/14 h) from 25 to 0℃, the OCR of the eels displayed a abrupt decrease up to 23℃, and between at 23 and 20℃, there was an agitation which showed a slight increase in the OCR with a duration of 1-2 days. Below 9℃, the OCR rhythm of the eels showed a constant state regardless of temperature decreasing. These results suggest that the Japanese eel has an upper incipient lethal temperature at 36℃, with a lower thermal limit at 9℃. The biochemical aspects of the eels influenced by water temperature need to be further studied.

Vg은 난생 척추동물의 성숙한 암컷 혈청에 존재하는 성 특이 단백질로서 에 의해 합성이 유도된다. 본 연구는 뱀장어 Vg과 ER 유전자 발현에 대한 androgen과 성장호르몬(GH)의 영향을 조사하였다. 미성숙 뱀장어()에 , 뱀장어 recombinant GH(eGH, ) 또는 methyltestosterone(MT, )를 각각 단독 또는 eGH 또는 MT와 혼합하여 주사한 후 10일 후에 샘플을 채취하였다. 간 ER과 Vg mRNA는 RT-PCR을