Table 1.
Timepoints of phenotype analysis.
Figure 1.
HA-TGF-β1(a) transgenic expression.
Panel A. Representative genomic DNA PCR screen of transgenic litters demonstrating germline transmission. Panel B. RT-PCR for transgene message in the prostate gland. GAPDH amplification serves as an internal control. Panel D. IHC of transgene demonstrates focal expression in ventral, lateral, and dorsolateral regions of secretory acini. Panel C. Wildtype acini demonstrate no immunoreactivity to HA epitope. Images C and D were captured at ×200 magnification.
Figure 2.
Expression of HA- TGF-β1(a) results in attenuation of prostate gland secretory epithelium.
Panels A and C. Representative micrographs of wild-type ventral prostate from 30-week and 62-week old mice, respectively. Micrographs shown are representative of all ages examined. Panel B. Overexpression of TGF-β1 results in thinning and denuding of the epithelial wall in a 29-week old transgenic mouse. Panel D. Ventral prostate from 62-week old transgenic mouse demonstrates severe attenuation. Panel F. IHC for collagen type IV demonstrates discontinuity of basement membrane in transgenic compared to wild-type (Panel E) prostate. Panel G. HA immunolocalization is observed in some areas of wall attenuation in transgenic mice. Panel H. Pyknotic appearing cells are evident in the lumen of acini in transgenic mice. Panel I. Quantitation of epithelial attenuation yields significantly greater thinning in aged transgenic compared to wild-type ventral prostate gland (P<0.05). Images A–F and H captured at ×200 magnification. Image G captured at ×400 magnification.
Figure 3.
Inflammation is increased in an age-dependent manner in nerve ganglia and associated vasculature of TGF-β1 transgenic mice.
Panel A. Vascular fibroplasia is evident in the wall of some vessels. Panel B. Focal regions of inflammation characterized by accumulations of immune cells is observed adjacent to some vessels in an age-dependent manner. Panel C and D. Peripheral inflammation appears within most prostatic interlobar parasympathetic ganglia and neurovascular bundles in transgenic mice. Panels E and F. Quantitation yields significantly greater inflammation in aged transgenic compared to wild-type vessels and ganglia, respectively (P<0.05). All images captured at ×200 magnification.
Figure 4.
Fibroplasia in transgenic mice exhibits markers of reactive stroma.
Serial sections showing stromal thickening (Panel A, arrow) with increased deposition of tenascin-C (Panel B, IHC for tenascin-C), consistent with a reactive stroma phenotype in areas surrounding some attenuated epithelial acini. Panel C. Tenascin-C immunoreactivity is also deposited near the base of stromal micronodules. Panel D and E. Tenascin-C deposition appears to form lamellar layers near the base and periphery of stromal micronodules. Panel F. Masson's trichrome staining indicates nodules are composed of collagen (blue). Images A–D & F were captured at ×200 and image E at ×400 magnification.
Figure 5.
Collagenous micronodules in TGF-β1 transgenic mice.
Panel A. Micronodules originate from the stroma immediately adjacent to epithelial acini (asterisk). Panel B. Micronodules are associated with a fibroplastic reactive stroma at their base (arrows) and project into the wall of epithelial acini. Panel C. Periacinar stroma is thickened in regions adjacent to micronodules. Vessels are often observed at the base of stromal micronodules (arrow). Panel D and E. Larger stromal nodules show greater deposition of matrix and project fully into the lumen of collapsed atrophic acini. Panel F. In certain cases, a collagenous micronodule (asterisk) appears adjacent to regions of transgene expression in the epithelial layer (arrowheads: IHC for HA epitope). Panel G. Focal immunoreactivity for HA epitope can be observed in epithelium adjacent to regions of periacinar stromal thickening in transgenic mice. Panel H. A significant elevation in general fibroplasia is noted in transgenic mice over one year of age as compared to age-matched wildtype controls and transgenic mice under one year of age (* P<0.05). Images A–F captured at ×200 magnification and image G at ×400.
Figure 6.
Recruitment of immature myeloid cells to parasympathetic ganglia is increased in TGF-β1 transgenic mice.
Panels A and B. Inflammatory lesions demonstrate significantly greater recruitment of CD115+ myeloid cells to parasympathetic neurovascular bundles in transgenic mice (Panel B) compared to wildtype control (Panel A). Panels C and D. Quantitation yields significantly greater CD115+ monocytes in transgenic compared to wildtype parasympathetic ganglia in mice over one year of age (* P<0.05) (Panel C), whereas no significant differences in F4/80 staining is observed (Panel D). All images captured at ×200.
Figure 7.
Tissue recombinants demonstrate TGFβ1-induced extracellular matrix deposition and inflammation.
Panel A. Recombination of UGM plus NHPrE1.2 vs. NHPrE1.2-HA-TGF-β1(a) demonstrates a decrease in graft size at both 8 and 12 weeks in recombinants that express HA-TGF-β1(a). Panels B and C. Trichrome staining demonstrates increased collagen production in HA-TGF-β1(a) expressing human prostate epithelia compared to vector control. Panels D and E. An increase in F4/80 positive macrophages is apparent in recombinants expressing HA-TGF-β1(a). Panels F and G. In addition, p65 activation is observed in both the epithelium and stroma in recombinants expressing HA-TGF-β1(a). Panels H and I. Control recombinants at 12 weeks maintain glandular acini structure and overall morphology, whereas recombinants made with HA-TGF-β1(a) epithelium exhibit atrophic and attenuated acini. Images B-I captured at ×100.