Lipopolysaccharide disrupts gap junctional intercellular communication in an immortalized ovine luteal endothelial cell line. (October 2019)

• Record Type: Journal Article
• Authors: Gram, Aykut; Grazul-Bilska, Anna T.; Boos, Alois; Rahman, Nafis A.; Kowalewski, Mariusz P.
• Is Part Of: Toxicology in vitro. Volume 60(2019)
• Page Start: 437
• Page End: 449
• Publication Date: 2019-10
• Subjects: LPS -- Luteal endothelial cells -- Connexins; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2019.05.017
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Gram-negative bacteria, in particular Escherichia coli with its cell wall lipopolysaccharide (LPS), often cause metritis and mastitis in domestic animals. Ovarian LPS accumulation may initiate local inflammatory reactions mediated through cell surface Toll-like receptors (TLRs). This may disrupt ovarian functionality leading to infertility. Possible adverse effects of LPS on luteal activity are not yet well explored. We hypothesized that LPS could lead to alterations in luteal vascular functionality. Therefore, we established an in vitro cell line model (OLENDO) by immortalizing microvascular endothelial cells isolated from ovine corpus luteum (CL) with a potent Simian Virus 40 T-antigen (SV40-Tag). OLENDO exhibit endothelial cell characteristics, like low-density lipoprotein (LDL) uptake, express BSL-I, and VEGFR2, as well as TLR2 and TLR4 receptors. LPS-treatment of OLENDO altered in vitro tube formation, had no effects on cell viability and decreased gap junctional intercellular communication (GJIC). LPS did not impair GJA1/Cx43 protein expression, but altered its cellular localization showing signs of internalization. Taken together, we demonstrated the mechanisms underlying LPS induced impairment of luteal GJIC and immune processes in a novel and well-characterized OLENDO cell line. Highlights: OLENDO cell line express specific endothelial cells markers, e.g., BSL-I and VEGFR2. OLENDO cells express connexins typical for endothelial cells (Cx37, Cx40 and Cx43), and TLR2 and TLR4 receptors. ICAM 1 is induced in OLENDO cells after LPS treatment. LPS disrupts in vitro formation of capillary-like structures and endothelial barrier function in OLENDO cells.