Immunohistochemical markers for domestic cat (Felis catus) ovarian tissue
(2019) f-TALES Ghent: Challenges in Reproductive Medicine — Location: Ghent, Belgium (18.September.2019)
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- et. al.
- Abstract
- Most species in the Felidae family are at risk of extinction, with the exception being the domestic cat, which has become an excellent model for reproductive studies due to its phylogenetic proximity to other felids. Several assisted reproduction techniques have been developed to aid in the conservation of these species. However, the lack of detailed knowledge of folliculogenesis in felids has limited the success of such techniques. Immunohistochemistry is a valuable tool for this type of research, but there are currently no commercially available species-specific markers. The aim of this study was therefore to identify, among existing markers, those that could be suitable to evaluate feline ovarian tissue. Ovaries containing follicles at different developmental stages from five adult female cats were collected, fixed in 4% paraformaldehyde and adequately processed, after which the following immunohistochemical assays were performed: GDF-9 (Goat anti-human GDF-9, mAb, Santa Cruz, Ref. Sc-12244), anti-Müllerian hormone (AMH - AMH anti-human mouse, mAb, clone [5/6], Serotec, ref MCA2246), α-inhibin (Rabbit anti-CD117 polyclonal ab, Dako, ref A4502), Ki67 (Ki67 anti-human monoclonal mouse , clone MIB 1, mAb, DAKO, ref. M7240), Kit-Ligand (KL - SCF anti-human mouse, mAb, clone G-3, Santa Cruz, ref. sc-13126), c-kit (Rabbit anti-CD117 polyclonal ab, Dako, ref A4502) and CD31 (PECAM-1 human clone EP3095, mAb, Epitomics, Ref 2530-1). A total of 3426 follicles (2706 primordial, 276 primary, 207 secondary and 237 antral) were analyzed, of which 99.5% were morphologically normal. GDF-9 was present in the oocyte cytoplasm at all stages of follicular development and AMH was detected in granulosa cells of primary, secondary and antral follicles, while inhibin-α was detected in mural cells of antral follicles, but not in cumulus cells. Ki67 showed granulosa cell proliferation in primary, secondary and antral follicles, whereas CD-31 was efficient for marking blood vessels in the ovarian stroma. On the other hand, markers KL and c-kit were not efficient for ovarian tissue in cats. The obtained results demonstrate the successful use of readily available markers to evaluate follicular viability before and after assisted reproduction techniques, and to ultimately expand the knowledge of folliculogenesis in this species.
- Affiliations
Universidade de Brasília Instituto de Ciências Biológicas
Citations
de Miranda Vasconcellos Vilela, J., & et al. (2019). Immunohistochemical markers for domestic cat (Felis catus) ovarian tissue. f-TALES Ghent: Challenges in Reproductive Medicine, Ghent, Belgium. https://hdl.handle.net/2078.5/106649