Developmental effects related to angiogenesis and osteogenic differentiation in Salmon larvae continuously exposed to dioxin-like 3,3',4,4'-tetrachlorobiphenyl (congener 77).

We have studied the effects of dioxin-like 3,3',4,4'-tetrachlorobiphenyl (PCB-77) on developmental effects related to angiogenesis and osteogenesis during early life-stages of salmon. Larvae were kept at 6°C and continuously exposed to waterborne PCB-77 (1 or 10 ng/L) initiated at the egg stage or 416-day degrees (dd) and throughout yolk-sac stage (716 dd) and for a total duration of 50 days (or 300 dd). Gene transcription analysis was performed on whole larvae total RNA at 548, 632, 674 and 716 dd using real-time PCR. Bone morphogenetic protein (bmp2 and bmp4), transforming growth factor β (TGF-β), estrogen receptors (ERα and ERβ), runx2, sox9 and collagen type 2 alpha 1 (col2a1) and vascular endothelial growth factor (VEGF) genes were studied. Effect on VEGF gene transcription was related to observation of heart rate, arrhythmia and anemia, demonstrating effects on vascular system development. Alizarine-red staining and quantification of ossified bone structures showed that PCB-77 produced concentration-dependent increases in the rate of osteogenic tissue formation. PCB-77 produced increases in col2a1 and runx2 transcription with subsequent induction of chondrogenesis and osteogenesis, respectively. The transcription of TGF-β gene was associated with ERβ transcription. Transcripts of AhR gene battery were differentially modulated by PCB-77 and these effects were dependent on concentration and larval age. Evidence of vascular system disruption by PCB-77 was observed as cardiac edema, anemia and arrhythmia in exposed individuals and as decreased level of VEGF gene transcription at early age. In general, our data indicate that PCB-77 produced developmental effects related to angiogenesis and osteogenic differentiation and disruption of vascular system development.