Immune Cell Profiling in the Ovarian Stroma of a Viviparous Fish during the Breeding Season: A Histological and Immunohistochemical Investigation

Abstract

The molly fish is a member of viviparous teleosts that are characterized by the fusion of the right and left ovaries during their early embryonic development. This fusion results in a singular and saccular ovary, where the germinal epithelium lines the internal lumen. The present study aimed to identify the immune cells in the ovarian stroma of Molly fish during the breeding season using histological and immunohistochemical analysis. Histological examination of the ovaries displayed oocytes at all different stages of development and degeneration. The ovocoel, a lymph-filled space, remains in the center of the ovary and branches posteriorly, creating the lumen of the gonoduct. The ovarian wall is composed of three layers: the mesothelium, tunica albuginea, and germinal epithelium. The developing ova were held together by the stroma, which consisted of vascular collagenous connective tissue clustered with immune cells. Immunohistochemical analysis revealed the presence of clusters of macrophages expressing APG5, IL-1β, TGF-β, S100, NF-κB, CD68, Iba-1, and Ach. Monocytes demonstrated positive immunoreactivity for both APG5 and IL-1β, whereas dendritic cells expressed only APG5. Furthermore, rodlet cells exhibited immunoreactivity for S100 protein, IL-1β, NF-κB, CD68, Nrf2, Ach, myostatin, SOX9, and Iba-1. In contrast, stem cells displayed immunoreactivity for Nrf2, myostatin, and SOX9. In conclusion, the ovarian stroma of Molly fish demonstrated a notable presence of immune cells, indicating their active involvement in immune reactions.