Thyroid hormone regulates the cell cycle inhibitor p27Kip1 in postnatal murine sertoli cells

Abstract

Thyroid hormone regulates early postnatal Sertoli cell proliferation. Transient neonatal hypothyroidism allows prolonged postnatal Sertoli cell mitogenesis and doubles adult Sertoli cell numbers, testis weight, and sperm production. The mechanism of this effect is unknown. Cell proliferation is stimulated by cyclins and cyclin-dependent kinases and inhibited by cyclin-dependent kinase inhibitors (CDKIs). T3 regulates the CDKI p27Kip1 in other cell types, and mice lacking p27Kip1 have increased testis size. To test the hypothesis that T3 regulates Sertoli cell mitogenesis by acting through p27Kip1, we compared expression of p27Kip1 in Sertoli cells of testes from euthyroid, hypothyroid, and hyperthyroid mice. At postnatal d 5-25, testes were collected and immunostained for p27Kip1 expression, or Sertoli cells were isolated enzymatically and used for p27Kip1 Western blotting, p27Kip1 immunostaining was low in rapidly proliferating 5-d-old Sertoli cells but had increased strongly in nonproliferating 25-d-old Sertoli cells, p27Kip1 immunostaining was reduced in Sertoli cells from hypothyroid mice compared with euthyroid controls at 10 and 16 d, consistent with increased Sertoli cell proliferation in these mice. Western blotting corroborated the p27Kip1 immunostaining, and p27Kip1 expression was greater in Sertoli cells from control compared with hypothyroid mice at postnatal d 10 and 16, but p27Kip1 expression was comparable by d 25. Hyperthyroidism increased p27Kip1 immunostaining relative to controls, and Western analysis indicated that Sertoli cells from 10-d-old hyperthyroid mice expressed more p27Kip1 than control mice. These results indicate that thyroid hormone status affects p27Kip1 expression in neonatal Sertoll cells, suggesting that T3 effects on Sertoli cell proliferation may be mediated through this CDKI.