Figure 1.
MIS-Smad2DN mice develop inclusion cysts with cells following a staining pattern similar to OSE.
Ovaries removed from 3-month-old MIS driven Smad2DN transgenic mice were fixed, processed and analyzed. H&E stained tissue demonstrated that the cells lining the cysts morphologically resembled the normal TEC due to ciliation. Immunohistochemical analysis showed that cells lining the cysts expressed CK8 and acetylated tubulin but lacked expression of the normal oviductal markers E-Cadherin, OVGP1 and PAX8. Black arrows, blue arrows and red arrows indicate cells lining the inclusion cysts, OSE and TEC respectively. Scale bars represent 100 µm.
Figure 2.
Intrabursal injection of AdCreGFP, but not intraoviductal injection, modifies ovarian phenotype.
Fluid-filled bursal cysts and degenerate ovaries were observed in p53flox/flox mice 6 months after intrabursal injection of AdCreGFP (A-C). Black arrow indicates ovarian tissue and blue arrow indicates oviductal tissue. The TEC showed normal expression of CK8 (D), PAX8 (E), and OVGP1 (F) both three and six months after intraoviductal viral injection. Scale bar represents 100 µm.
Table 1.
Summary of Tumor Incidence.
Figure 3.
Intrabursal or intraoviductal injection of adenovirus results in OSE or TEC specific infection.
Immunohistochemical and immunofluorescent detection of GFP and Cre-recombinase indicates tissue specific infection. Intrabursal injection shows staining of the OSE with limited leakage to adjacent tissue including the bursa. Intraoviductal injection results in limited infection of the TEC. Scale bars represent 100 µm.
Figure 4.
High Smad2DN transgene expression on a FVB background results in teratoma formation.
The large teratomas contained many diverse structures including squamous cysts filled with layers of keratin (A), nervous tissue (B), bone formation (C), undifferentiated cells invading into an area of nervous tissue-like cells (D), thyroid follicle-like glands (E), and ciliated cells lining fluid-filled cysts (F). Immunohistochemical staining for CK8 was strongly positive in epithelial cells lining inclusion cysts (G) and infiltrating tumor cells (H). The OSE had an altered, papillary morphology and stained positive for epithelial marker CK8 (I). Acetylated tubulin marked the ciliated cells of the inclusion cysts as well as ovarian stroma (J). Abnormal OVGP1 expression was noted in the ovarian cortex obtained from an eight-week-old mouse (K). The teratomas lacked staining for phopsho-Smad2/3 (L). Scale bar represent 100 µm. Amplification of the Smad2DN by real time qPCR revealed significantly higher transgene expression in mice with the teratoma phenotype as compared to the mice with only the ovarian inclusion cysts phenotype (M).