Immunohistochemical Staining of IL‐1 Alpha and IL‐1 Receptor Antagonist but not IL‐1 Beta in Cultures of Sertoli Cells

Abstract

The interleukin-1 (IL-1) system has been suggested to be involved in the cell cell cross talk within the testis. To identify a testicular cell source of IL-1 alpha, IL-1 beta and IL-1 receptor antagonist (IL-1ra), immature mouse Sertoli cells were isolated, purified, cultured and examined for the cellular compartment localization of these cytokines by immunohistochemical staining. Our results show that both Germ cells and Sertoli cells in unpurified Sertoli cell cultures (before hypotonic shock) and purified culture of Sertoli cells (after hypotonic shock) were stained for IL-1 alpha. The levels of this cytokine were increased in Sertoli cells when the purified cultures were stimulated with lipopolysaccharide (LPS) (5 microg/mL). However, we could not identify a positive staining for IL-1 beta when Sertoli cell cultures were stained for this cytokine, even after stimulation with various concentrations of LPS (0.1-10 microg/mL). On the other hand, immunohistochemical staining of isolated Sertoli cells without treatment with hypotonic shock (cultures containing Sertoli cells and Germ cells) for IL-1ra showed constitutive positive staining of both cell types (Sertoli cells and Germ cells). Our results, using immunohistochemical staining, may indicate the different expression of IL-1 alpha, IL-1 beta and IL-1ra in Sertoli cells. These results may suggest the involvement of IL-1 system in the autocrine and paracrine regulation of testicular cell functions.

Effect of Interleukin-1 Receptor Antagonist Gene Deletion on Male Mouse Fertility

“…I IL-1 receptor was demonstrated (43). This report may indicate that IL-1, which is constitutively expressed (physiological levels) in testicular tissues (11)(12)(13)(14)(15)(16)(17)(18)(19)(20) and was shown to increase germ cell proliferation and DNA synthesis (44,45), is not a limiting factor in steroidogenesis and spermatogenesis under physiological conditions. It is possible that other cytokines (such as TNF-␣) may subjugate the role of IL-1.…”

“…Recently, we have demonstrated overexpression of IL-1␣, IL-1␤, and IL-1 Ra in homogenates of sexually immature mice, compared with mature mice (13). We also showed that mature human sperm cells express and secrete IL-1-like molecules under in vitro conditions (21,22), and that Sertoli cells and germ cells could produce IL-1 Ra under in vitro conditions (17,18). The levels of IL-1 Ra in Sertoli cells were induced by lipopolysaccharide (LPS), IL-1, and FSH (17,23).…”

“…We also showed that mature human sperm cells express and secrete IL-1-like molecules under in vitro conditions (21,22), and that Sertoli cells and germ cells could produce IL-1 Ra under in vitro conditions (17,18). The levels of IL-1 Ra in Sertoli cells were induced by lipopolysaccharide (LPS), IL-1, and FSH (17,23). IL-1 receptors have been identified, characterized, and localized in mouse testis (24,25).…”

“…In the interstitium, Leydig cells (13)(14)(15) and testicular macrophages (16) were found to produce and secrete IL-1, preferably the ␤-form. IL-1␣ production can be detected in tubular sources, namely Sertoli cells (13,14,(17)(18)(19) and germ cells (17,20). Recently, we have demonstrated overexpression of IL-1␣, IL-1␤, and IL-1 Ra in homogenates of sexually immature mice, compared with mature mice (13).…”

Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 5: Intercellular junctions and contacts between germs cells and Sertoli cells and their regulatory interactions, testicular cholesterol, and genes/proteins associated with more than one germ cell generation

“…IL-1a [ID: T5-931] production occurs in germ cells (Haugen et al, 1994) and residual bodies stimulate production of IL-1a by Sertoli cells (Wang et al, 1998). Germ cells produce IL-1Ra [ID: T5-1017] in vitro (Huleihel et al, 2001;Zeyse et al, 2000) and IL-1 receptors (e.g., [ID: T5-1018]) are present in mouse testis (Gomez et al, 1997;Takao et al, 1990). IL1b [ID: T5-1019] is found in the cytoplasm and nucleus of spermatogonia and spermatocytes, while IL-1Ra is in spermatogonia and in the Golgi apparatus (Huleihel et al, 2003).…”