Our results suggested that the environment of south China is suitable for the growth and reproduce of R. chensinensis. The main results were as follows: 1. The ratio of mating of adult R. chensinensis were 84% in south China, and the ratio of spawning were 68%, the survival rate were 42%, and the hatch rate of oosperm were 56%. 2. The temperature in south China is higher than that in east-north China, which reduces the time of the embryonic development of R. chensinensis. In natural condition, the early embryonic development of R. chensinensis required 131–155 h and the postembryonic development required 28– 30 days. Totally, the embryonic development required 32–35 days, which was less about one month than that in east-north China. 3. The higher temperature expedited the embryonic development, but it did not affect the normal development of embryo. During the period of embryonic development, there almost no unexpected died and misshapen individual. 4. The results of our study on the dissection of tadpole and larval frog and the organic slice of gonad confirm that the early development of gonad was in good condition. The processes of the emergence of original gonad and the split of germ cell etc were on the rails. 5. With the reduce of the time of embryonic development of R. chensinensis, its original gonad came into being ahead of time, but the differentiation of gonad was behind the schedule. 6. The pattern of gonadal sex differentiation of R. chensinensis is the differentiated type. 7. The results we had reported above not only enrich the development biology but also provide the foundation of theory and practice for introducing the R. chensinensis to south China. doi:10.1016/j.cbpc.2008.10.013 MOLECULAR CLONING AND EVOLUTIONARY ANALYSIS OF HYPOXIA-INDUCIBLE FACTOR 1 ALPHA, AND EPO RESPONSIVENESS UNDER HYPOXIA IN THE TIBETAN VERTEBRATES PANTHOLOPS HODGSONI, MYOSPALAX BAILEYI, MYOSPALAX CANSUS, MICROTUS OECONOMUS AND GYMNOCYPRIS PRZEWALSKII X.Q. Chen a,⁎ , Y.B. Cao a , Y. Wang a , Y. Liu a , X.C. Chen c , G. Cai d , P.H. Xiao d , J.Z. Du a,b,⁎ a Dept. Physiology, School of Medicine, Zhejiang University, Hangzhou 310058, China b Dept. Physiology, College of Life Sciences, Zhejiang University, Hangzhou 310058, China c Northwest Institute of Plateau Biology, The Chinese Academy of Sciences, Qinghai 810000, China d Qinghai Kekexili National Nature Reserve, Geermu, 816000, China E-mails: chewyg@zjuem.zju.edu.cn; dujz@zju.edu.cn Hypoxia-inducible factor 1α (HIF-1α) is an essential mediator of oxygen homeostasis. The Tibetan antelope (P. hodgsoni), plateau pika (O. curzoniae), plateau zokor (M. baileyi, M. cansus), root vole (M. oeconomus), and naked carp (G. przewalskii) are native vertebrates of the Qinghai–Tibetan plateau and well acclimatized to hypoxia. To better understand their adaptive mechanisms to hypoxia, cDNA encoding HIF-1α was isolated and characterized. The deduced HIF-1α sequences of P. hodgsoni, M. oeconomus, M. baileyi and M. cansus showed 90–99% identity with those of the human, rat, and yak; G. przewalskii had 57–89% identity with rainbow trout and common carp. The conservational and phylogenic clustering of the HIF-1α sequences was consistent with vertebrate classification. We estimated codons under positive selection. All positive selection sites were outside the key domain, but between the key domains of TAD-N and TAD-C (607L, 611T, 607E, 622D, 623E, 624L, 627V, 632M, 645T, and 663Q), and another was 8E in M. baileyi. The evidence shows that plateau animals have specializations for hypoxia linked to changes of Tibetan Plateau ecological environment. HIF-1 increased in the cortex and liver of the mice, M. baileyi, and M. oeconomus under hypoxia, but not in O. curzoniae; EPO increased in the cortex and kidney of mice under hypoxia, but only in the kidney of M. oeconomus. In addition, under CoCl 2 -hypoxia, EPO increased in the cortex and kidney of three Tibetan mammals, but not in mice. The differences in HIF and EPO between Tibetan animals and lowland mice suggest that diverse strategies are involved in hypoxia. HIF-1 may play a role in mice and M. baileyi but not in O. curzoniae. This work was supported by the NSFC (Major Project No. 30393130, and Nos.30470648 and 30570227) and the “973” Program (No. 2006CB504100). doi:10.1016/j.cbpc.2008.10.014 NF-KB FUNCTIONS THE DIVERSITY OF CELLULAR IGF-I/IGFBP-1 EXPRESSION BY HYPOXIA IN TIBETAN PLATEAU MAMMALS X.Q. Chen, S.J. Wang, Y. Liu, J.Z. Du Dept. Physiology, Divi. Neurobiology and Physiology, School of Medicine, Zhejiang University, Hangzhou 310058, China E-mail: dujz@zju.edu.cn Ochotona curzoniae, Microtus oeconomus and Myospalax baileyi are all native mammals that reside at Qinghai–Tibetan plateau in China and well acclimatized to environmental hypoxia. The present paper addresses the NF-kB's, a nuclear transcriptional factor, involvement in hypoxia stress-induced diversity of IGF-I/ IGFBP-1 expression in hepatic and brain cells of Tibetan Plateau mammals. The IGF-I/IGFBP-1 from the prefrontal cortex and the liver cells was tested 6 h after hypoxia exposure (by CoCl 2 injection i.p. 20, 40 mg/kg or by normobaric hypoxia, 16.0%, 10.8%, 8.0 %O 2 ) of the Plateau native mammals and mice. PDTC, an inhibitor of NF-κB, was used and preinjected before the hypoxia to evaluated NF-kB action. The results showed that 1) the IGF-I expression in mice hepatic cells of M. oeconomus and M. baileyi markedly increased after the hypoxia exposure, but there was no response in the liver of O.curzoniae; 2) the IGFBP-1 expression in mice hepatic cells of O. curzoniae and M. baileyi markedly enhanced, but no response occurred in M. oeconomus after the hypoxia; 3) PDTC pretreated before hypoxia reversed the hypoxia-enhanced IGF-I in M. oeconomus and M. baileyi; 4) PDTC treatment also reversed the hypoxia-enhanced IGFBP-1 in O. curzoniae and M. baileyi; 5) hypoxia increased the IGF-I mRNA in brain of M. oeconomus and O. curzoniae but not of mice; 6) hypoxia did not induce changes of IGF-I levels in the brain cells of both plateau mammals and laboratory mice. The data suggest that 1) different pattern in IGF-I/IGFBP-1 expression induced by hypoxia represents a diversities in hormone regulation and cell protection from damage in Tibetan native mammals; 2) NF-κB mediates the transcription of IGF-I/IGFBP-1 in liver cells subjecting to hypoxia; Together, the diversity of target-gene phenotype expression may contribute to the multi-model in cell protection from hypoxia damage. doi:10.1016/j.cbpc.2008.10.015 A TRANSCRIPTOME SCHEME OF GILL REVEALS THE COLD ACCLIMATION STRATEGIES IN ZEBRAFISH (DANIO RERIO) Ming-Yi Chou a,b , Chung-Der Hsiao a , Bo-Kai Liao a , Pung-Pung Hwang a,b a Institute of Cellular and Organismic Biology, Academia Sinica, Taiwan b Institute of Fishery Science, National Taiwan University, Taiwan E-mail: pphwang@gate.sinica.edu.tw Global analysis of gene expression using functional genomics ap- proaches is a revolutionizing technique for molecular physiological studies. DNA microarray is a powerful technique to identify the differential expressed genes among two or more populations as an indirect measure of functional changes of the related proteins. Fish are ectodermic vertebrates and have some advantages to study the relationships between environ- mental temperature and their genome. The purpose of this study was to investigate the effects of low temperature on the functions and differentiation of gill cells. We used microarray technology to compare the gene expression profiles in gill cells between acute and chronic treatments of low temperature. Adult zebrafish were acclimated to a low temperature of 12 °C for 1 (1-d) and 30 d (30-d), and the gene expression patterns of gill were compared to control groups (28 °C). The transcrip- tome profiles of 1-d and 30-d treatments were extreme different, only 2 genes were conserved, suggesting that zebrafish may utilize distinct strategies for short- and long-term cold acclimation. Several ionoregula- tion-related genes were identied from these profiles and we further investigated the role of ionoregulatory genes in zebrafish gills during cold acclimation. Broad functions of ionocytes related genes were induced by cold, indicating that terminal differentiation and function of ionocytes were stimulated to recover the cold-induced imbalance of ion and acid/ Abstracts 450