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Evidence that MMP-2 and TIMP-2 are at play in the FSH-induced changes in Sertoli cells
Juliette Longin1, Brigitte Le Magueresse-Battistoni
1Inserm U329, Hopital Debrousse, 29 rue soeur Bouvier, 69322, Cedex 05, Lyon, France.
Abstract:
FSH has been shown to elicit in vitro changes in the Sertoli cell cytoskeleton through involving proteases, and cytochalasin-D mimics FSH. Testis extracts were screened (RT-PCR) for various metalloproteinases (MMPs), 20-day-old rat Sertoli cells were purified, cultured and treated with FSH, cytochalasin D and TNFalpha (to antagonize FSH action). Cell shape (phase-contrast microscopy) and levels for MMP-2 (gelatin zymography) and its inhibitor TIMP-2 (Northern and Western blot) were monitored. TNFalpha-treated cells spread readily and grew larger than FSH-treated cells. Cytochalasin-D mimicked FSH, and MMP-2 production and TIMP-2 gene expression were augmented. Interestingly, TNFalpha reversed FSH- and cytochalasin D-induced effects both on cell shape and on MMP-2 and TIMP-2. These effects occurred during the first 48 h of culture, when Sertoli cells migrated from the freshly dispersed aggregates, but not once cells were organized in monolayers. MMP-2 and TIMP-2 are likely involved in the FSH-induced changes in Sertoli cells.
Insights
Follicle-stimulating hormone (FSH) influences Sertoli cell shape and migration via matrix metalloproteinase-2 (MMP-2) and its inhibitor TIMP-2. Tumor necrosis factor-alpha (TNFα) antagonizes these FSH-induced effects.
Area of Science:
- Reproductive Biology
- Cell Biology
- Molecular Endocrinology
Background:
- Follicle-stimulating hormone (FSH) is known to induce in vitro changes in the Sertoli cell cytoskeleton.
- Cytochalasin-D, a known cytoskeletal disruptor, mimics FSH effects.
- Proteases are implicated in FSH-mediated cytoskeletal alterations.
Purpose of the Study:
- To investigate the role of matrix metalloproteinases (MMPs) in FSH-induced Sertoli cell changes.
- To examine the expression of MMP-2 and its inhibitor TIMP-2 in response to FSH.
- To determine the effect of TNFα on FSH-mediated Sertoli cell morphology and MMP/TIMP levels.
Main Methods:
- Screening of testis extracts for MMPs using RT-PCR.
- Purification and culture of 20-day-old rat Sertoli cells.
- Treatment of Sertoli cells with FSH, cytochalasin D, and TNFα.
- Monitoring of cell shape via phase-contrast microscopy.
- Quantification of MMP-2 levels using gelatin zymography.
- Assessment of TIMP-2 gene and protein expression via Northern and Western blot.
Main Results:
- TNFα treatment led to increased Sertoli cell spreading and size compared to FSH treatment.
- Cytochalasin-D mimicked FSH effects, augmenting MMP-2 production and TIMP-2 gene expression.
- TNFα reversed the effects of FSH and cytochalasin-D on cell shape, MMP-2, and TIMP-2.
- These dynamic changes were observed during initial Sertoli cell migration (first 48 hours) but not in established monolayers.
Conclusions:
- Matrix metalloproteinase-2 (MMP-2) and its inhibitor TIMP-2 are likely involved in FSH-induced alterations of Sertoli cell cytoskeleton and migration.
- FSH action on Sertoli cell morphology and MMP/TIMP expression can be modulated by TNFα.
- The dynamic interplay between FSH, MMP-2, and TIMP-2 is crucial during the early migratory phase of Sertoli cells.
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