Transgenerational effects of the endocrine disruptor vinclozolin on the prostate transcriptome and adult onset disease

Abstract

Purpose: The ability of an endocrine disruptor exposure during gonadal sex determination to promote a transgenerational prostate disease phenotype was investigated in the current study.

Methods: Exposure of an F0 gestating female rat to the endocrine disruptor vinclozolin during F1 embryo gonadal sex determination promoted a transgenerational adult onset prostate disease phenotype. The prostate disease phenotype and physiological parameters were determined for males from F1 to F4 generations and the prostate transcriptome was assessed in the F3 generation.

Results: Although the prostate in prepubertal animals develops normally, abnormalities involving epithelial cell atrophy, glandular dysgenesis, prostatitis, and hyperplasia of the ventral prostate develop in older animals. The ventral prostate phenotype was transmitted for four generations (F1-F4). Analysis of the ventral prostate transcriptome demonstrated 954 genes had significantly altered expression between control and vinclozolin F3 generation animals. Analysis of isolated ventral prostate epithelial cells identified 259 genes with significantly altered expression between control and vinclozolin F3 generation animals. Characterization of regulated genes demonstrated several cellular pathways were influenced, including calcium and WNT. A number of genes identified have been shown to be associated with prostate disease and cancer, including beta-microseminoprotein (Msp) and tumor necrosis factor receptor superfamily 6 (Fadd).

Conclusions: The ability of an endocrine disruptor to promote transgenerational prostate abnormalities appears to involve an epigenetic transgenerational alteration in the prostate transcriptome and male germ-line. Potential epigenetic transgenerational alteration of prostate gene expression by environmental compounds may be important to consider in the etiology of adult onset prostate disease.

Fig. 1

Ventral prostate morphology from P15 control (A) and vinclozolin (B) and P30 control (C,E) and vinclozolin (D,F) F2 generation rats. (A–D) Magnification 200× and (E,F) magnification 400×. Arrows indicate ductal epithelium. Asterisks (*) indicate prostate ducts with atrophic/degenerated epithelial cells. Micrographs are representative of 4 controls and 8 vinclozolin P15 samples and 12 control and 14 vinclozolin P30 samples.

Fig. 2

The percent of animals affected containing atrophic ducts in the ventral prostate sections from F2 generation control (gray bars) and vinclozolin (black bars) from postnatal day P15 (control n =4 and vinclozolin n =8), P30 (control n =12 and vinclozolin n =14), P120(control n =10 and vinclozolin n =10), and P300 (control n =21 and vinclozolin n =23) rats. Asterisks (*) indicate statistically significant differences from control by Fisher’s Exact test.

Fig. 3

Ventral prostate morphology from P120 control (A,C,E) and vinclozolin (B,D,F) F2 generation rats. Ventral prostate cross-sections of distal (A,B), intermediate (C,D), and proximal (E,F) regions. Asterisks (*) indicate atrophic ducts. Magnification 100× (A–D)and200× (E,F). Micrographs are representative of 10 controls and 10 vinclozolin samples.

Fig. 4

Ventral prostate morphology from control (A,B) and vinclozolin (C–F) P300 F3 generation rats. F: Regions of epithelial cell hyperplasia. Asterisks (*) indicate atrophic ducts. Arrows denotes region of vacuoles in the basal epithelium (E). Magnification 100× (A,C,F) and 1000× (B,D,E). Micrographs are representative of 5 controls and 12 vinclozolin samples; except (F), which is 1 control and 4 vinclozolin samples.

Fig. 5

Lateral (A,B) and dorsal (C,D) prostate morphology from control (A,C) and vinclozolin (B,D) P300 F3 generation rats. Magnification 200×. Micrographs are representative of 10 controls and 12 vinclozolin samples.

Fig. 6

Dendrogram analyses of the microarray regulated gene data from whole ventral prostate tissue(A)and isolated epithelial cells(B)from F3 generation rats. Dendrogram were produced in Gene spring using an unsupervise cluster analysis. Dendrogram comprised from 954 regulated gene list from ventral prostate tissue (A) and 259 regulated gene list from isolated epithelial cells (B). Green color indicates a decrease in expression and red color indicates an increase in expression, scale provided.

Fig. 7

Functional categorization of the regulated gene lists. Each gene was separated into a functional family and graphed to indicate numbers of genes up- and down-regulated.(A) 954 gene list from ventral prostate tissue and (B) 259 epithelial list.

Fig. 8

Functional connectivity analyses of the common 55 regulated gene lists between total prostate and epithelial cells. Cellular processes associated with each gene in the 55 list were determined based on the number of arrows connected to each box (connectivity). Shown are only the boxes that had more then 5 connections with genes from the 55 common regulated gene list. The shaded genes listed are not in the 55 gene list but provide bridging connections, while the white genes listed are in the 55 gene list.