RESUMEN
Neuropeptide Y receptor Y8 (NPY8R) is a fish-specific receptor with two subtypes, NPY8AR and NPY8BR. Changes in expression levels during physiological processes or in vivo regulation after ventricular injection suggest that NPY8BR plays an important role in feeding regulation; this has been found in only a few fish, at present. In order to better understand the physiological function of npy8br, especially in digestion, we used clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology to generate npy8br-/- Japanese medaka (Oryzias latipes). We found that the deletion of npy8br in medaka larvae affected their feeding and digestion ability, ultimately affecting their growth. Specifically, npy8br deficiency in medaka larvae resulted in decreased feed intake and decreased expression levels of orexigenic genes (npy and agrp). npy8br-/- medaka larvae fed for 10 d (10th day of feeding) still had incompletely digested brine shrimp (Artemia nauplii) in the digestive tract 8 h after feeding, the messenger RNA (mRNA) expression levels of digestion-related genes (amy, lpl, ctra, and ctrb) were significantly decreased, and the activity of amylase, trypsin, and lipase also significantly decreased. The deletion of npy8br in medaka larvae inhibited the growth and significantly decreased the expression of growth-related genes (gh and igf1). Hematoxylin and eosin (H&E) sections of intestinal tissue showed that npy8br-/- medaka larvae had damaged intestine, thinned intestinal wall, and shortened intestinal villi. So far, this is the first npy8br gene knockout model established in fish and the first demonstration that npy8br plays an important role in digestion.
Asunto(s)
Digestión , Técnicas de Inactivación de Genes , Larva , Oryzias , Receptores de Neuropéptido Y , Animales , Oryzias/genética , Receptores de Neuropéptido Y/genética , Larva/genética , Sistemas CRISPR-Cas , Conducta Alimentaria , Proteínas de Peces/genética , Proteínas de Peces/metabolismoRESUMEN
Intracellular bacteria such as those belonging to the genus Edwardsiella can survive and proliferate within macrophages. However, the detailed mechanisms underlying the host macrophage immune response and pathogen evasion strategies remain unknown. To advance the field of host macrophage research, we successfully established transgenic (Tg) Japanese medaka Oryzias latipes that possesses fluorescently visualized macrophages. As a macrophage marker, the macrophage-expressed gene 1.1 (mpeg1.1) was selected because of its predominant expression across various tissues in medaka. To validate the macrophage characteristics of the fluorescently labeled cells, May-Grünwald Giemsa staining and peroxidase staining were conducted. The labeled cells exhibited morphological features consistent with those of monocyte/macrophage-like cells and tested negative for peroxidase activity. Through co-localization studies, the fluorescently labeled cells co-localized with E. piscicida in the intestines and kidneys of infected medaka larvae, confirming the ingestion of bacteria through phagocytosis. In addition, the labeled cells expressed macrophage markers but lacked a neutrophil marker. These results suggested that the fluorescently labeled cells of Tg[mpeg1.1:mCherry/mAG] medaka were monocytes/macrophages, which will be useful for future studies aimed at understanding the mechanisms of macrophage-mediated bacterial infections.
Asunto(s)
Animales Modificados Genéticamente , Edwardsiella , Infecciones por Enterobacteriaceae , Enfermedades de los Peces , Macrófagos , Oryzias , Fagocitosis , Animales , Oryzias/genética , Macrófagos/microbiología , Macrófagos/metabolismo , Macrófagos/inmunología , Edwardsiella/genética , Infecciones por Enterobacteriaceae/veterinaria , Infecciones por Enterobacteriaceae/inmunología , Infecciones por Enterobacteriaceae/microbiología , Enfermedades de los Peces/microbiología , Enfermedades de los Peces/inmunología , Larva/microbiología , Larva/genética , Larva/inmunologíaRESUMEN
Nectins are adhesion molecules that play a crucial role in the organization of epithelial and endothelial junctions and function as receptors for the entry of herpes simplex virus. However, the role of Nectin4 remains poorly understood in fish. In this study, nectin4 gene was cloned from medaka (OlNectin4). OlNectin4 was located on chromosome 18 and contained 11 exons, with a total genome length of 25754 bp, coding sequences of 1689 bp, coding 562 amino acids and a molecular weight of 65.5 kDa. OlNectin4 contained four regions, including an Immunoglobulin region, an Immunoglobulin C-2 Type region, a Transmembrane region and a Coiled coil region. OlNectin4 shared 47.18 % and 25.00 % identity to Paralichthys olivaceus and Mus musculus, respectively. In adult medaka, the transcript of nectin4 was predominantly detected in gill. During red spotted grouper nervous necrosis virus (RGNNV) infection, overexpression of OlNectin4 in GE cells significantly increased viral gene transcriptions. Meanwhile, Two mutants named OlNectin4â³4 (+4 bp) and OlNectin4â³7 (-7 bp) medaka were established using CRISPR-Cas9 system. Nectin4-KO medaka had higher mortality than WT after infected with RGNNV. Moreover, the expression of RGNNV RNA2 gene in different tissues of the Nectin4-KO were higher than WT medaka after challenged with RGNNV. The brain and eye of Nectin4-KO medaka which RGNNV mainly enriched, exhibited significantly higher expression of interferon signaling genes than in WT. Taken together, the OlNectin4 plays a complex role against RGNNV infection by inducing interferon responses for viral clearance.
Asunto(s)
Enfermedades de los Peces , Proteínas de Peces , Nectinas , Nodaviridae , Oryzias , Infecciones por Virus ARN , Animales , Oryzias/genética , Oryzias/inmunología , Nodaviridae/fisiología , Infecciones por Virus ARN/veterinaria , Infecciones por Virus ARN/inmunología , Nectinas/genética , Nectinas/inmunología , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/química , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Filogenia , Secuencia de Aminoácidos , Inmunidad Innata/genética , Alineación de Secuencia/veterinaria , Regulación de la Expresión Génica/inmunología , Perfilación de la Expresión Génica/veterinariaRESUMEN
The cytokine interleukin (IL)-22 has been identified in several fish species; however, its functional significance in the gills of these fish species remains unclear. In this study, we analyzed the expression of proinflammatory cytokines, antimicrobial peptides, and IL-22 binding protein in the gills of wild-type and IL-22-knockout (IL-22 KO) medaka under dextran sulfate sodium-induced inflammation. We also produced medaka recombinant IL-22 (rIL-22) and analyzed the expression of immune-related genes in rIL-22-stimulated primary cell cultures from gills. The il1b, il6, tnfa, and hamp genes were significantly upregulated in wild-type gills upon dextran sulfate sodium stimulation compared with the naïve state but not in IL-22 KO gills. il22bp transcripts were barely detectable in the IL-22 KO medaka gills. However, the expression of il1b, il6, hamp, and il22bp was upregulated in rIL-22-stimulated gill cell culture. These results suggest IL-22 could be involved in immune responses through inflammatory cytokine and antimicrobial peptide production in fish gills.
Asunto(s)
Oryzias , Animales , Oryzias/genética , Branquias , Sulfato de Dextran , Interleucina-6 , Interleucinas/genética , Citocinas , Expresión Génica , Interleucina-22RESUMEN
pax6 is a canonic master gene for eye formation. Knockout of pax6 affects the development of craniofacial skeleton and eye in mice. Whether pax6 affects the development of spinal bone has not been reported yet. In the present study, we used CRISPR/Cas9 system to generate Olpax6.1 mutant in Japanese medaka. Phenotype analysis showed that ocular mutation caused by the Olpax6.1 mutation occurred in the homozygous mutant. The phenotype of heterozygotes is not significantly different from that of wild-type. In addition, knockout Olpax6.1 resulted in severe curvature of the spine in the homozygous F2 generation. Comparative transcriptome analysis and qRT-PCR revealed that the defective Olpax6.1 protein caused a decrease in the expression level of sp7, col10a1a, and bglap, while the expression level of xylt2 did not change significantly. The functional enrichment of differentially expressed genes (DEGs) using the Kyoto Encyclopedia of Genes and Genomes database showed that the DEGs between Olpax6.1 mutation and wild-type were enriched in p53 signaling pathway, extracellular matrix (ECM) -receptor interaction, et al. Our results indicated that the defective Olpax6.1 protein results in the reduction of sp7 expression level and the activation of p53 signaling pathway, which leads to a decrease in the expression of genes encoding ECM protein, such as collagen protein family and bone gamma-carboxyglutamate protein, which further inhibits bone development. Based on the phenotype and molecular mechanism of ocular mutation and spinal curvature induced by Olpax6.1 knockout, we believe that the Olpax6.1-/- mutant could be a potential model for the study of spondylo-ocular syndrome.
Asunto(s)
Oryzias , Animales , Ratones , Oryzias/genética , Oryzias/metabolismo , Ratones Noqueados , Proteína p53 Supresora de Tumor/genética , MutaciónRESUMEN
The mutual relationship between the intestinal immune system and the gut microbiota has received a great deal of attention. In mammals, interleukin-17A and F (IL-17A/F) are inflammatory cytokines and key regulators of the gut microbiota. However, in teleosts, the function of IL-17A/F in controlling the gut microbiota is poorly understood. We attempted to elucidate the importance of teleost IL-17 signaling in controlling gut microbiota. We previously established a knockout (KO) of IL-17 receptor A (RA) 1, a receptor for IL-17A/F, in the Japanese medaka (Oryzias latipes) using the CRISPR-Cas9 system and performed 16S rRNA-based metagenomic analyses using the anterior and posterior sections of the intestinal tract. The number of observed OTUs in the anterior intestine was significantly decreased in IL-17RA1 KO medaka compared to that in the wild-type (WT). Furthermore, ß-diversity analysis (weighted UniFrac) revealed considerably different bacterial composition in the anterior intestine of IL-17RA1 KO compared to WT, with similar findings in α-diversity. Notably, the pathogen Plesiomonas shigelloides was significantly increased in the posterior intestine of IL-17RA1 KO medaka. These findings indicate that signaling via IL-17RA1 is required to maintain a healthy gut microbiota in teleosts and mammals. The involvement of IL-17RA1 in controlling the gut microbiota has been demonstrated, resulting in microbiome dysbiosis in IL-17RA1 KO medaka.
Asunto(s)
Microbiota , Oryzias , Animales , Intestinos , Oryzias/genética , ARN Ribosómico 16S/genética , Receptores de Interleucina-17/genéticaRESUMEN
In mammals, interleukin (IL)-17 receptor C (IL-17RC) and IL-17RA mediate IL-17A and IL-17F signaling to produce mucin, antimicrobial peptides, and maintain healthy intestinal flora. However, IL-17RC signaling in fish remains unclear. In this study, three il17rc transcripts (il17rca1, il17rca2, and il17rcb) from the Japanese medaka (Oryzias latipes) were cloned; il17rca1 and il17rca2 mRNAs were alternatively spliced from il17rca pre-mRNA as transcript variants. The il17rca and il17rcb genes were located on chromosomes 7 and 5, respectively. Teleost clades containing medaka il17rca and il17rcb clustered separately from the tetrapod clade. In adult tissues, il17rca1 expression was significantly higher than il17rca2 and il17rcb. Conversely, il17rcb expression was significantly higher in embryos and larvae. These expression patterns changed following infection with Edwardsiella piscicida and Aeromonas hydrophila. Furthermore, an immunoprecipitation assay using recombinant IL-17RCs and rIL-17A/Fs suggested that, in teleosts, three ligands could function in signaling through two IL-17RCs.
Asunto(s)
Interleucina-17/metabolismo , Oryzias/inmunología , Receptores de Interleucina-17/metabolismo , Aeromonas hydrophila/fisiología , Empalme Alternativo , Animales , Mapeo Cromosómico , Edwardsiella/fisiología , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Expresión Génica/inmunología , Ligandos , Oryzias/genética , Oryzias/microbiología , Filogenia , Receptores de Interleucina-17/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Sintenía , Distribución Tisular/inmunologíaRESUMEN
MicroRNAs (miRNAs) are important gene expression regulators implicated in many biological processes, but we lack a global understanding of how miRNA genes evolve and contribute to developmental canalization and phenotypic diversification. Whole-genome duplication events likely provide a substrate for species divergence and phenotypic change by increasing gene numbers and relaxing evolutionary pressures. To understand the consequences of genome duplication on miRNA evolution, we studied miRNA genes following the teleost genome duplication (TGD). Analysis of miRNA genes in four teleosts and in spotted gar, whose lineage diverged before the TGD, revealed that miRNA genes were retained in ohnologous pairs more frequently than protein-coding genes, and that gene losses occurred rapidly after the TGD. Genomic context influenced retention rates, with clustered miRNA genes retained more often than nonclustered miRNA genes and intergenic miRNA genes retained more frequently than intragenic miRNA genes, which often shared the evolutionary fate of their protein-coding host. Expression analyses revealed both conserved and divergent expression patterns across species in line with miRNA functions in phenotypic canalization and diversification, respectively. Finally, major strands of miRNA genes experienced stronger purifying selection, especially in their seeds and 3'-complementary regions, compared with minor strands, which nonetheless also displayed evolutionary features compatible with constrained function. This study provides the first genome-wide, multispecies analysis of the mechanisms influencing metazoan miRNA evolution after whole-genome duplication.
Asunto(s)
Evolución Biológica , Peces/genética , Genoma , MicroARNs/genética , Animales , Secuencia de Bases , Secuencia Conservada , Peces/metabolismo , Duplicación de Gen , Gónadas/metabolismo , Familia de Multigenes , Selección Genética , Especificidad de la EspecieRESUMEN
Gonad development and histopathological changes typically associated with endocrine disruption were evaluated in female Japanese medaka (Oryzias latipes) exposed to river water from four representative cross-sections in the Yellow River (YR), China. Fish were held in the river water treatments from fertilization. Advanced ovarian development was observed in fish exposed to river water from Qinhe cross-section at 20 days post-hatch (dph) and in fish exposed to river water from all four cross-sections at 60 dph. Histopathological changes including increased oocyte atresia, perifollicular cell hyperplasia/hypertrophy, changes in ovarian staging, interstitial fibrosis and interstitial proteinaceous fluid were observed in the gonads of fish at 60 dph after exposure to river water from some cross-sections. Cytoplasmic retraction and karyoplasmic clumping were observed in fish exposed to river water from all four cross-sections at 60 dph. The results indicate that development and reproductive function in Yellow River fish is impaired, placing fish populations at risk.
Asunto(s)
Oryzias , Contaminantes Químicos del Agua , Animales , China , Femenino , Gónadas , Agua , Contaminantes Químicos del Agua/toxicidadRESUMEN
ASC is a component of the inflammasome playing crucial roles in the inflammatory response. In mammals, ASC induces pyroptosis and inflammatory cytokine production. In this study, three asc genes (asc1, asc2, and asc3) from the Japanese medaka (Oryzias latipes) were identified and characterized. These asc genes were tandem replicates on chromosome 16, and their exon-intron structures differed between them. All three ASCs conserved the pyrin and caspase-recruitment domains, which are important for inflammasome formation. In phylogenetic analysis, all ASCs clustered with those of other teleosts. The asc1 expression levels were significantly higher in several organs than those of asc2 and asc3, suggesting that asc1 may act as a dominant asc in the Japanese medaka. Expression of the three asc genes showed different patterns during Aeromonas hydrophila and Edwardsiella piscicida infections. Furthermore, their expression was adequately down-regulated in the medaka fin-derived cells stimulated with ATP for 12 h, while asc2 expression was statistically up-regulated after nigericin stimulation for 24 h. Moreover, the expression of asc2 and asc3 was significantly higher in the skin of ASC-1-knockout medaka than in that of the wild type medaka during A. hydrophila infection.
Asunto(s)
Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Duplicación de Gen/inmunología , Oryzias/inmunología , Aeromonas hydrophila/inmunología , Animales , Animales Modificados Genéticamente , Línea Celular , Edwardsiella/inmunología , Enfermedades de los Peces/genética , Enfermedades de los Peces/microbiología , Proteínas de Peces/metabolismo , Regulación de la Expresión Génica/inmunología , Técnicas de Inactivación de Genes , Inflamasomas/genética , Oryzias/genética , Oryzias/microbiología , Filogenia , PiroptosisRESUMEN
Apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) is a component of inflammasome, which plays crucial roles in the inflammatory response. In mammals, ASC regulates caspase-1 activation, thereby inducing pyroptosis and producing activated inflammatory cytokines. In addition, ASC also interacts with receptor-interacting protein kinase 2 (RIPK2) and induces nuclear factor-κB (NF-κB) activation. However, the role of ASC remains poorly understood in fish. In this study, we focused on elucidating the role of ASC in fish that were infected with Aeromonas hydrophila using Japanese medaka (Oryzias latipes) as fish model, and ASC-knockout (KO) medaka was established using CRISPR-Cas9 system. ASC-KO and wild type (WT) medakas were infected with A. hydrophila, and mortality was observed. ASC-KO medaka demonstrated higher mortality than WT. Moreover, the expression of immune-related genes in the kidney and intestine of the ASC-KO and WT medakas challenged with A. hydrophila were analyzed. Following A. hydrophila infection, the kidney of ASC-KO medaka exhibited significantly lower expression of NF-κB regulated genes (e.g., IL-1ß, IL-6, IL-8 and TNF-α) and RIPK2 gene than in WT kidney. Moreover, to investigate the immune response against A. hydrophila via ASC in the medaka, bacterial burden, superoxide anion production, and lactate dehydrogenase release in the kidney cells of ASC-KO medaka were measured. After infection, these responses in ASC-KO medaka were significantly decreased compared to those in WT. These results suggest that the medaka ASC plays a critical role against A. hydrophila infection by inducing inflammatory responses and cell death for bacterial clearance.
Asunto(s)
Proteínas del Citoesqueleto/genética , Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Infecciones por Bacterias Gramnegativas/veterinaria , Inflamasomas/inmunología , Oryzias , Aeromonas hydrophila/fisiología , Animales , Proteínas del Citoesqueleto/metabolismo , Enfermedades de los Peces/microbiología , Proteínas de Peces/metabolismo , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/microbiología , Interacciones Huésped-Patógeno , Inflamasomas/genéticaRESUMEN
The growing use of octocrylene (OC) in sunscreens has posed a great threat to aquatic organisms. In the present study, to assess its reproductive toxicity and mechanism, paired Japanese medaka (Oryzias latipes) (F0) were exposed to OC at nominal concentrations of 5, 50, and 500 µg/L for 28 d. Significant increases were observed in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of F0 medaka at 500 µg/L OC (p < 0.05) without significant differences in fecundity. The fertility was significantly decreased at all treatments (p < 0.05). Significant increases in the percent of mature oocytes were observed at 5 and 500 µg/L OC, in which contrary to the percent of spermatozoa (p < 0.05). The plasma sex hormones and vitellogenin levels significantly increased in males at all treatments and in females at 50 and 500 µg/L OC (p < 0.05). In addition, the levels of fshß and lhß in the brains and the levels of fshr, lhr and cyp17α in the gonads were significantly upregulated in males at all treatments (p < 0.05), in line with those of ar, erα, erß and cyp19ß in the brains of male and female. The upregulation of vtg in male and female livers was observed only at 500 µg/L OC and upregulation of star and hsd3ß was observed in testis at all treatments (p < 0.05). Continued exposure to OC significantly induced increases in the time to hatching, morphological abnormality rates, and cumulative death rates of F1 embryos, inconsistent with body length of F1 larvae (p < 0.05). Therefore, the responses of the exposed fish at the biochemical and molecular levels indicated reproductive toxicity and estrogenic activity of OC, providing insights into the mechanism of OC.
Asunto(s)
Oryzias , Contaminantes Químicos del Agua , Acrilatos , Animales , Estrógenos , Femenino , Hígado , Masculino , Reproducción , VitelogeninasRESUMEN
In mammals, interleukin 17 (IL-17), which is produced mainly by Th17 cells, is a hallmark inflammatory cytokine that plays key roles in the protection against infection and intestinal mucosal immunity. The mammalian IL-17 receptor family comprises five members (IL-17RA-E). Of these, IL-17RA is important in the control of the bacterial microbiota in mucosal tissues, particularly in the intestine, where it acts as a receptor for IL-17A and -F. In this study, the nucleotide sequence of IL-17RA1 cDNA from Japanese medaka (Oryzias latipes) of the Cab strain was determined and compared to two IL-17RA cDNAs (i.e., IL-17RA1 and IL-17RA2) of Japanese medaka Hd-rR strain downloaded from NCBI. Hd-rR 17RA1 and IL-17RA2 were located on chromosome 23 and chromosome 6, respectively, and phylogenetic tree analysis revealed that teleost IL-17RA1 and IL-17RA2 were separated in different clusters. Synteny analysis revealed that Japanese medaka IL-17RA1 and mammalian IL-17RA are conserved. IL-17RA1 expression levels in the gills, intestine, whole kidney, skin, and spleen were significantly higher than those of IL-17RA2, suggesting that IL-17RA1 is an important functional receptor in mucosal immunity. Interestingly, the expression levels of both IL-17RA genes were notably higher in the posterior than in the anterior intestinal tract section. Furthermore, despite its lower basal expression, IL-17RA2 expression was significantly increased at 72â¯h post Edwardsiella tarda infection.
Asunto(s)
Cromosomas , Proteínas de Peces , Regulación de la Expresión Génica , Oryzias , Receptores de Interleucina-17 , Animales , Cromosomas/genética , Cromosomas/metabolismo , Proteínas de Peces/biosíntesis , Proteínas de Peces/genética , Oryzias/genética , Oryzias/metabolismo , Receptores de Interleucina-17/biosíntesis , Receptores de Interleucina-17/genéticaRESUMEN
Numerous cytosolic DNA sensors (CDSs), which are very important for recognizing cytosolic dsDNA derived from intracellular viruses and bacteria, exist in mammals. However, teleost CDSs are poorly understood. In this study, four CDSs, including the cyclic GMP-AMP synthase (cGAS), Sm-like protein 14 homolog A (LSm14A), DEAH-box helicase (DHX) 9, and DHX36 genes were identified in Japanese medaka, Oryzias latipes, and their expression patterns were elucidated. The expression of these genes was upregulated in the intestines and kidney of CpG-ODN-stimulated medaka. The cGAS and LSm14A genes were significantly induced in the intestines, kidney, and spleen of formalin-killed Edwardsiella tarda-treated medaka; the DHX9 and DHX36 genes were not. cGAS gene expression was induced only in the intestines of live E. tarda-treated medaka. These results suggest that the transcription of four CDS genes of medaka responds to dsDNA stimulation, and cGAS is probably more important for the immune response against E. tarda infection.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Peces/genética , Regulación de la Expresión Génica , Oryzias/genética , Receptores de Reconocimiento de Patrones/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de Unión al ADN/inmunología , Proteínas de Unión al ADN/metabolismo , Edwardsiella tarda/inmunología , Proteínas de Peces/inmunología , Proteínas de Peces/metabolismo , Regulación de la Expresión Génica/inmunología , Oligodesoxirribonucleótidos/inmunología , Oryzias/clasificación , Oryzias/inmunología , Filogenia , Receptores de Reconocimiento de Patrones/inmunología , Receptores de Reconocimiento de Patrones/metabolismo , Distribución TisularRESUMEN
Estrogenic activities of river water from four representative cross-sections of the Yellow River (Zhengzhou section) and their effects on reproduction and development of fish were assessed. MVLN assay showed estradiol equivalents of river water from Yiluohe, Xinmanghe, Qinhe and Huayuankou cross-sections were 1.09 ± 0.11, 0.72 ± 0.01, 1.19 ± 0.19 and 0.80 ± 0.04 ng/L, respectively. Significant vitellogenin (VTG) inductions were observed in adult male Japanese madaka (Oryzias latipes) after 30 days of exposure to river water from Yiluohe and Qinhe cross-sections (p < 0.05). Hepatic-somatic index was significantly elevated in fish exposed to water from Qinhe cross-section (p < 0.05). A significant delay in time to hatching was observed in embryos treated by water from Xinmanghe cross-section (p < 0.05). Significant lower survivals were observed in fish treated by water from Yiluohe and Xinmanghe cross-sections after a full life cycle exposure (p < 0.05). Exposure of water from Yiluohe and Qinhe cross-sections induced significantly elevated VTG levels in the first sexually mature male fish (p < 0.05). Both the in vitro and in vivo bioassay demonstrate endocrine disrupting chemicals exist in the Yellow River (Zhengzhou section) and fish in Yiluohe and Qinhe cross-sections can be at a risk of reproductive and developmental impairment.