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BLTK1 murine Leydig cells: a novel steroidogenic model for evaluating the effects of reproductive and developmental
Agnes L Forgacs1, Qi Ding, Rosemary G Jaremba
1Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan 48824, USA.
Abstract:
Leydig cells are the primary site of androgen biosynthesis in males. Several environmental toxicants target steroidogenesis resulting in both developmental and reproductive effects including testicular dysgenesis syndrome. The aim of this study was to evaluate the effect of several structurally diverse endocrine disrupting compounds (EDCs) on steroidogenesis in a novel BLTK1 murine Leydig cell model. We demonstrate that BLTK1 cells possess a fully functional steroidogenic pathway that produces low basal levels of testosterone (T) and express all the necessary steroidogenic enzymes including Star, Cyp11a1, Cyp17a1, Hsd3b1, Hsd17b3, and Srd5a1. Recombinant human chorionic gonadotropin (rhCG) and forskolin (FSK) elicited concentration- and time-dependent induction of 3',5'-cyclic adenosine monophosphate, progesterone (P), and T, as well as the differential expression of Star, Hsd3b6, Hsd17b3, and Srd5a1 messenger RNA levels. The evaluation of several structurally diverse male reproductive toxicants including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), atrazine, prochloraz, triclosan, monoethylhexyl phthalate (MEHP), glyphosate, and RDX in BLTK1 cells suggests different modes of action perturb steroidogenesis. For example, prochloraz and triclosan antifungals reduced rhCG induction of T, consistent with published in vivo data but did not alter basal T levels. In contrast, atrazine and MEHP elicited modest induction of basal T but antagonized rhCG-mediated induction of T levels, whereas TCDD, glyphosate, and RDX had no effect on basal or rhCG induction of T in BLTK1 cells. These results suggest that BLTK1 cells maintain rhCG-inducible steroidogenesis and are a viable in vitro Leydig cell model to evaluate the effects of EDCs on steroidogenesis. This model can also be used to elucidate the different mechanisms underlying toxicant-mediated disruption of steroidogenesis.
Insights
This study introduces a new BLTK1 mouse Leydig cell model to test how environmental toxicants affect male hormone production. The model successfully identified different ways endocrine disrupting compounds disrupt testosterone synthesis.
Area of Science:
- Endocrinology
- Toxicology
- Cell Biology
Background:
- Leydig cells are crucial for male androgen biosynthesis.
- Environmental toxicants can disrupt steroidogenesis, leading to reproductive issues.
- A novel BLTK1 murine Leydig cell model was developed to study these effects.
Purpose of the Study:
- To evaluate the impact of diverse endocrine disrupting compounds (EDCs) on steroidogenesis using the BLTK1 Leydig cell model.
- To characterize the steroidogenic pathway and responsiveness to stimulation in BLTK1 cells.
- To elucidate the distinct mechanisms by which various toxicants affect Leydig cell function.
Main Methods:
- Utilized a novel BLTK1 murine Leydig cell line.
- Assessed basal and stimulated testosterone (T) and progesterone (P) production.
- Measured messenger RNA (mRNA) expression of key steroidogenic enzymes.
- Exposed cells to structurally diverse male reproductive toxicants including TCDD, atrazine, prochloraz, triclosan, MEHP, glyphosate, and RDX.
Main Results:
- BLTK1 cells possess a functional steroidogenic pathway, producing basal T and expressing necessary enzymes.
- Recombinant human chorionic gonadotropin (rhCG) and forskolin (FSK) stimulated steroidogenesis and altered mRNA levels.
- Different toxicants exhibited varied effects: prochloraz/triclosan inhibited rhCG-stimulated T, atrazine/MEHP modulated basal and stimulated T, while TCDD, glyphosate, and RDX had no effect.
- The study identified distinct modes of action for toxicant-induced disruption of steroidogenesis.
Conclusions:
- The BLTK1 cell model is a viable in vitro system for evaluating EDC effects on steroidogenesis.
- This model allows for the elucidation of mechanisms underlying toxicant-mediated disruption of Leydig cell function.
- The findings highlight the diverse impacts of environmental toxicants on male reproductive health.
