共查询到20条相似文献，搜索用时 250 毫秒
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Steroids are known to play a crucial role in gonadal sex differentiation in many non-mammalian vertebrates, but also in the
gonadal sex change of hermaphroditic teleosts. We investigated the expression of two genes encoding key steroidogenic enzymes,
i.e., cytochrome P450 aromatase (P450arom) and cytochrome P45011β-hydroxylase (P45011β), during the sex change of the protogynous
rice field eel, Monopterus albus. Using RT-PCR with degenerate primers, we cloned rice field eel homologous fragments for both genes (rcP450arom and rcP45011β)
as indicated by the high level of homology with P450arom and P45011β sequences from various vertebrates. Gonadal expression
of rcP450arom and rcP45011β mRNA levels were then assessed during the sex change by semi-quantitative RT-PCR and a real-time
RT-PCR. rcP450arom was predominantly expressed in ovary, much less in ovotestis, and barely in testis. Conversely, P45011β
was markedly up-regulated at the onset of testicular development. These findings underline that regulation of steroidogenesis
is an important process in the sex change of protogynous rice field eel, and they clearly indicate that the concomitant down-regulation
of P450arom and up-regulation of P45011β are of pivotal importance to the sex change of this species. 相似文献
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The effects of water temperature on the development of hermaphroditic gonads in red seabream (Pagrus major) and on mRNA expression of cytochrome P450 aromatase (P450arom) and 11β-hydroxylase were examined. High water temperature suppressed both expression of P450arom and 11β-hydroxylase and the development of oocytes in ovarian portion of hermaphroditic gonads. 相似文献
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黄鳝脑芳香化酶基因cDNA的克隆及组织表达特异性分析 总被引：1，自引：0，他引：1
根据NCBI数据库报道的黄鳝芳香化酶基因的gDNA序列,设计了一对特异性引物.用Trizol试剂盒提取黄鳝脑总RNA,反转录获得cDNA第一条链,进行PCR扩增.扩增产物经过回收、连接、测序后得到了一条长1514 bp的芳香化酶基因cDNA序列.同源性比较表明该基因属于脑型P450aromB,与其他鱼类脑型P450aromB的同源性较高(>70%),与性腺型P450aromA的同源性较低(<60%),与自身性腺型芳香化酶同源性为59.5%.采用RT-PCR的方法对该基因在雄性黄鳝的各组织表达情况进行了分析,结果表明,该基因的转录本较高的表达于脑和精巢中,较低的在皮肤中表达,而在肝脏、心脏、小肠、肌肉等组织中没有检测到该基因的表达. 相似文献
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J. K. Sundaray K. Ohta A. Yamaguchi T. Kitano M. Matsuyama 《Fish physiology and biochemistry》2005,31(2-3):137-141
Pseudolabrus sieboldi, wrasse being a diurnal spawner provides a good opportunity to study the endocrine mechanism of estrogen formation in brain
and gonads. Moreover, an extremely large amount of E2 was produced in serum and testis of wrasse. It is assumed that the presence
of E2 may play a major role in diurnal gametogenesis in male fish. In this study brain type aromatase have been isolated,
cloned and sequenced from the brain of wrasse. Further, the expression pattern of brain type aromatase in gonads and adult
tissue of male and female fish have been analyzed. In addition, the diurnal expression pattern of brain type aromatase in
both male and female fish brain during spawning season have been analyzed.
The P450arom (br) was isolated, cloned and sequenced from both male and female bamboleaf wrasse. The P450arom (br) gene (1877
sequenced nucleotide) contains an ORF of 1470 bp, a 5′-UTR of 18 bp and at least 407 bp in 3′-UTR. The amino acid sequence
homology in the coding region of wrasse P450arom (br) is high compared to that of medaka, Oryzias latipes (80%), rainbow trout type 2, Oncorhynchu mykiss (78.2%), fugu, Takifugu ribripes (78%) rainbow trout type 1, (76%), goldfish, Carassius auratus (66.8%) and zebrafish, Danio rerio (66.2%). Expression study reveals that P450arom (br) mRNA were most abundant in brains of both male and female fish throughout
the day during the spawning season. RT-PCR study revealed that P450arom (br) was expressed in skin, anal fin and tail fin
of both male and female wrasse. P450arom (br) was not detected at any time of the spawning day in either ovary or testis of
wrasse. 相似文献
6.
Shigeho Ijiri Norio Takei Shinji Andachi Kohei Yamauchi 《Fish physiology and biochemistry》2003,28(1-4):209-210
Antibodies against P450scc, P450c17 and P450arom were generated using recombinant proteins. In eel testis, P450scc and P450c17 were immunolocalized as clusters in Leydig cells. In vitellogenic eel ovary, P450scc and P450c17 immunoreactive cells were localized as clusters in the outer layer of the ovarian follicle. In contrast, P450arom seemed to be immunolocalized in the innermost follicle layer. 相似文献
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The effect of gonadotropins, forskolin and IGF-I on steroidogenic enzyme gene expression and E2 production by the rainbow trout follicle were measured. The results indicate that 3β-HSD gene expression is regulated by gonadotropins partially through the cAMP/PKA mechanism. The effect of IGF-I on P450arom mRNA levels suggests that IGF-I is a major regulator of P450arom gene expression. 相似文献
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在水温分别为20(常温对照组)、25、30℃条件下,采用RT-PCR方法,研究性腺成熟期雄性黄颡鱼组织中P450芳香化酶(P450aromA和P450aromB)的mRNA表达水平,同时测定性腺成熟系数(GSI)、肝重指数(HSI)、肥满度(CF)和血浆睾酮(T),雌二醇(E2)含量。结果表明,P450aromA在性腺发育成熟期雄性黄颡鱼心脏、肝、脾、胃、肾脏、精巢、鳃、脑、头肾、肠管组织中均无表达,P450aromB只在脑和肠中有表达,在脑中表达量高于肠,表达量随着水温升高而显著下降;各组实验鱼血浆T和E2含量与芳香化酶基因表达量存在相关关系。该实验结果表明,P450aromA可能与精子发生相关,可能不参与黄颡鱼精子排放过程;P450aromB在雄鱼脑中高度表达,可能参与性腺成熟期精子活力保持与排放过程,且应激温度越高,对性腺成熟期精子活力保持与排放过程影响越大。 相似文献
10.
In the present study, we implanted 2‐year‐old female red‐spotted grouper, Epinephelus akaara, with a non‐steroidal aromatase inhibitor (AI), fadrozole, in the breeding season and examined changes in gonadal histology, serum sex steroids, aromatase activities and P450 aromatase (P450arom) gene expression in gonads after AI implantation. Aromatase inhibitor at doses from 0.1 to 10.0 mg kg?1 BW induced a sex inversion and completion of spermatogenesis up to the functional male phase, but doses of 1.0 and 10.0 mg kg?1 BW AI produced more males than 0.1 mg kg?1 BW AI. Serum estradiol‐17β (E2) levels decreased, but 11‐ketotestosterone (11‐KT) levels increased significantly in all the AI‐implanted groups, whereas testosterone (T) levels increased significantly only in the 1.0 mg kg?1 BW AI‐implanted group. Aromatase activities and P450arom gene expression in gonads were inhibited significantly in the AI‐implanted groups, which was in accordance with the decrease in serum E2 levels. These results suggested the optimal dose of AI to induce sex inversion to be 1.0 mg kg?1 BW. Furthermore, the sex inversion induced by AI may be attributed to the inhibition of P450arom gene expression and aromatase activity and the resultant decrease in the biosynthesis of endogenous E2. Meanwhile, the elevated 11‐KT levels were also associated closely with the occurrence of sex inversion in protogynous red‐spotted grouper. 相似文献
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Yasuhisa Kobayashi Tomoki Sunobe Tohru Kobayashi Yoshitaka Nagahama Masaru Nakamura 《Fish physiology and biochemistry》2005,31(2-3):123-127
Despite numerous endocrine studies on sex change in teleost, no general mechanism that mediates sex change has emerged. The
gobiid fish, Trimma okinawae, can change sex in both directions repeatedly. This phenomenon of sex change in goby assigns it as an excellent animal model
to elucidate the understanding mechanisms of sex change. In hermaphrodite fishes, estrogen plays a particularly important
role in natural and experimentally induced sex change. To investigate the role of estrogen in the serial-sex changing fish
T. okinawae, we cloned and analyzed the 5′-flanking regions of P450arom genes from goby genome DNA. Both regions have consensus sequences
of TATA, CRE and ERE. Ad4 binding site was restricted in the region of P450aromA. These findings indicate that different regulators
control the expression of the two P450arom genes. 相似文献
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Koichiro Gen Sonoko Yamaguchi Koichi Okuzawa Naoki Kumakura Hideki Tanaka Hirohiko Kagawa 《Fish physiology and biochemistry》2003,28(1-4):77-80
The duality of gonadotropins (GTHs), follicle-stimulating hormone (FSH) and luteinizing hormone (LH), has been confirmed in most teleost species, but very little is known about their biological functions. To elucidate the physiological roles of FSH and LH in fish reproduction, the expression profiles of GTH subunit genes during gonadal development were analyzed in both male and female red seabream. Furthermore, in vitro studies were carried out to examine the effects of GTHs on steroid hormone production and cytochrome P450 aromatase (P450arom) expression in red seabream gonads. In both sexes, LHβ mRNA was maintained at high levels from the early gametogenesis until spawning season, and declined with gonadal regression. Interestingly, FSHβ mRNA levels in males increased in parallel with testicular development, whereas those in female were remained low throughout oocyte development. From in vitro studies using purified red seabream FSH and LH, both GTHs had a similar potency in stimulating 11-ketotestosterone production by testicular slices, while the biological activity of FSH was much lower than that of LH in stimulating production of estradiol-17β by vitellogenic follicles. Moreover, expression of P450arom mRNA was induced by LH, but not FSH, in ovarian follicles in vitro. FSH was also ineffective in inducing maturational competence and final oocyte maturation. These results suggest that, unlike salmonids, FSH may play an important role during gametogenesis in male, but not female, red seabream, whereas LH may be involved in regulation of both early and late gametogenesis in both sexes. 相似文献
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条斑星鲽CYP19a基因克隆及其在雄鱼生殖周期中的表达 总被引：1，自引：0，他引：1
细胞色素P450芳香化酶(P450arom)作为P450细胞色素酶超家族中的一员,是性类固醇生成途径中的末端酶,能够将雄激素转化为雌激素。在大多数脊椎动物中,P450arom由CYP19单基因编码。但是在鱼类中存在两种P450芳香化酶,分为性腺型芳香化酶(P450aromA)和脑型芳香化酶(P450aromB),它们由不同的基因(CYP19a和CYP19b)编码,分布在不同的组织。通过简并引物扩增及RACE cDNA扩增克隆,在国内外首次获得全长为2167bp的条斑星鲽CYP19a cDNA序列,并将推测的氨基酸序列与其它物种P450arom氨基酸序列进行多重比较,发现存在跨膜螺旋区、I-螺旋区、Ozol肽区、芳香化酶特异保守区以及血红素结合区。通过RT-PCR分析了P450aromA mRNA在条斑星鲽不同组织中的表达情况,结果表明,CYP19a基因主要在脑、卵巢和精巢中表达,其次在肠、肝脏、肾脏也有少量表达。同时也分析了P450aromA mRNA在处于不同发育期的精巢中的表达情况,发现在Ⅱ期精巢中表达量最高,在Ⅴ期精巢中表达量最低。 相似文献
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Jin GX Wen HS He F Li JF Chen CF Zhang JR Chen XY Shi B Shi D Yang YP Qi BX Li N 《Fish physiology and biochemistry》2012,38(3):807-817
P450c17, a key steroidogenic enzyme, plays important roles in the production of sex steroid and cortisol. In teleost, there are two types of P450c17, P450c17-I possessing 17α-hydroxylase and 17, 20-lyase activities, and P450c17-II only possessing 17α-hydroxylase activity. This work describes the molecular cloning of the cDNA encoding the barfin flounder (Verasper moseri) P450c17-I and P450c17-II by means of RT-PCR and 5' and 3' rapid amplification of cDNA ends (RACE) analyses and mRNA expression profiles analyzing by semiquantitative RT-PCR. Respectively, P450c17-I and P450c17-II mRNA levels in the testes correlated with serum testosterone (T) level, as well as gonadosomatic index (GSI) of males during specific stages of spermatogenesis. P450c17-I and P450c17-II mRNA were expressed in the testis and ovary, suggesting that both of them participate in the production of sex steroid in barfin flounder gonads. P450c17-I mRNA was undetectable; in contrast, P450c17-II mRNA was detected at the highest level in the head kidney, meaning that only P450c17-II is involved in the production of cortisol in barfin flounder. The results demonstrated that both of P450c17-I and P450c17-II participate in the production of sex steroid in male barfin flounder gonads. 相似文献
17.
Anders Goksøyr Tommy Andersson Donald R. Buhler John J. Stegeman David E. Williams Lars Förlin 《Fish physiology and biochemistry》1991,9(1):1-13
Antibodies prepared against the major -naphthoflavone (BNF)-inducible cytochrome P450 (P450) forms from three species of fish (rainbow trout, Atlantic cod, and scup) well separated in teleost phylogeny, were used to investigate the immunochemical relatedness of liver microsomal P450 in different species of BNF-treated fish and rat. Rabbit polyclonal IgG against all three P450s and mouse monoclonal antibodies prepared against scup P450E were employed in this study. Liver microsomes were prepared from BNF-treated specimens of hagfish, herring, rainbow trout, cod, scup, perch, plaice and rat. With Western blotting it was shown that the various antibodies cross-reacted with a protein band in liver microsomes in the P450-region of each of the BNF-treated fish species. The apparent molecular weight of the cross-reacting proteins showed differences within the range 54,000–59,000 daltons. The effects of the different antibodies on the microsomal BNF-inducible 7-ethoxyresorufin O-deethylase (EROD) activity gave inhibition patterns that reflected to a certain extent the phylogenetic relationship of the species investigated. In rat microsomes a protein band of relative molecular mass similar to rat P450c (Mr=54,000) was recognized by all antibodies. In addition, a second band of lower molecular mass was strongly recognized by anti-cod P450c antibodies, and faintly stained with anti-rainbow trout P450LM4b IgG and anti-scup P450E MAb 1-12-3. This band could correspond to rat P450d, the isosafrole-inducible rat isoenzyme. Considering the long separate evolutionary history of some of these fishes (50–200 million years), the results demonstrate that certain antigenic epitopes in the BNF-inducible P450 isoenzymes have been strongly conserved during the evolution of fish species. These conserved epitopes seem however not to be directly involved in the measured EROD activities. Furthermore, the results suggest that the BNF-inducible P450s in fish contain regions with structural similarity to the homologous counterpart that has evolved through gene duplication into a P450 family in mammals containing at least two gene products (the P450IA gene family). 相似文献
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Cytochrome P450c17 (CYP17, 17α‐hydroxylase/17,20‐lyase) is a critical enzyme in the production of androgens and estrogens in vertebrates. A 2102 bp full‐length cDNA of P450c17‐II (CYP17A2) has been isolated from the ovary of half‐smooth tongue sole, Cynoglossus semilaevis which encodes 524 amino acids. The putative P450c17‐II enzyme shares higher sequence identity with those of teleosts than with P450c17‐I of vertebrate. The similarity between the two types of tongue sole P450c17 was 48%.Semi‐quantitative RT‐PCR analysis of spatial expression showed the enzyme was specifically expressed in the ovary and the head kidney. However, temporal expression shows that P450c17‐II can be found in the brain. Furthermore, temporal expression pattern of P450c17‐II in ovary and brain revealed developmental stage‐dependency, and ovary P450c17‐II expressed remarkably throughout the whole reproductive cycle. Otherwise, the expression pattern of P450c17‐II in head kidney indicated negative ovary development‐dependence. In addition, combined with our data on P450c17‐I, T and E2 levels, the results further endorse the critical role of P450c17‐II during shift in steroidogenesis, suggesting that P450c17‐I and ‐II may act together to this physiological process. Based on the present study, we indicate an important role for P450c17‐II during ovarian development. 相似文献
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P.H. Strobl-Mazzulla G.C. López L.A. Miranda G.M. Somoza 《Fish physiology and biochemistry》2003,28(1-4):51-52
The molecular cloning of a cDNA encoding the pejerrey brain cytochrome P450 aromatase (P450aromB) is described. This form shares higher identity to other brain aromatases than with their respective ovarian counterparts and the self ovarian aromatase. Tissue-specific expression of both aromatases was examined in pejerrey by RT-PCR. The immunocytochemical distribution of P450aromB was described. 相似文献
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Malin Celander Donald R. Buhler Lars Förlin Anders Goksøyr Cristobal L. Miranda Bruce R. Woodin John J. Stegeman 《Fish physiology and biochemistry》1996,15(4):323-332
Multiple P450 proteins have been purified from several teleost species, including rainbow trout (Oncorhynchus mykiss), scup (Stenotomus chrysops) and Atlantic cod (Gadus morhua). Identity, relationships and/or functions have been established in these fish species for the cytochrome P4501 As. Information about the structure, function, regulation and relationships of other piscine cytochrome P450 (CYP) proteins is sparse. In the present study we have focused on constitutively expressed CYP forms, P450con and LMC5 isolated from rainbow trout, P450A from scup, and P450b from Atlantic cod, and we consider evidence for the relationship of these proteins to mammalian members of the CYP3A subfamily. Reciprocal western blot analysis shows that P450con and LMC5, isolated from rainbow trout in two different laboratories, are closely related and ostensibly identical proteins. These trout proteins show specific reciprocal cross-reactivity with scup P450A, and polyclonal antibodies (PAb) to the trout and scup proteins both recognize cod P450b, indicating that rainbow trout P450con/LMC5, scup P450A and cod P450b are immunochemically-related proteins. In analyses of liver microsomes of trout, scup and cod, PAb to trout P450con/LMC5 and scup P450A recognize only bands that are identical in migration to the CYP proteins purified from these species, and which were used as immunogens. These CYP proteins purified from fish are each immunochemically-related to mammalian CYP3A proteins, showing recognition by PAb to human CYP3A4 and to rat CYP3A1. PAb to the mammalian CYP3As also recognize the same bands in liver microsomes from these fish species as seen by PAb to the fish proteins. These results strongly suggest that these fish proteins are members of theCYP3 gene family and probably theCYP3A subfamily. Although sequence analysis is required before their designation in the CYP3A subfamily can be confirmed and specified, we refer to these as CYP3A-like. Immunoblot analyses of hepatic microsomes from other fish species with PAb to scup P450A and trout P450con show that multiple CYP3A-like proteins are expressed in liver of several species, including killifish (Fundulus heteroclitus) and winter flounder (Pleuronectes americanus). Important questions still remain to be addressed concerning CYP3A structure, multiplicity, physiological function, regulation and metabolism of endogenous as well as exogenous substrates in fish.Part of this study was presented at the 10th International Symposium on Microsomes and Drug Oxidations. Toronto, Canada, July 18–21, 1994. 相似文献