Abstract
The glucose transporters (GLUTs) gene encode glycoproteins responsible for facilitating transfer of glucose across plasma membrane. In testis, different members of this family are present. In particular the main GLUT mRNA expression within the adult testis is the type 8, while type 1 is more expressed in prepubertal testis. Thyroid hormone, which receptors and function have been characterized in the testis, plays a crucial role in the cellular energetic metabolism. In fact, in the immature Sertoli cells, GLUT1 is up regulated by l-triiodothyronine (T(3)). The aim of this paper is to investigate the expression profile of GLUT1 and GLUT8 in the testis during development and in adulthood and analyse the role of T(3) on their expression. To analyse the expression of GLUT8 and GLUT1 we performed Northern blot and RT-PCR experiments in the whole testis and in Sertoli cells from rats of different ages. Treatments in vivo and in vitro with T(3) were used to study the effect of thyroid hormones on GLUT1 and GLUT8 expression. The activity of the rat GLUT1 promoter and its regulation by T(3) was studied with transient transfections in gonadal and non-gonadal cell lines and in primary Sertoli cell cultures. GLUT8 is expressed at a low level in the prepubertal testis and Sertoli cells and does not appear to be under T(3) control. GLUT1 is the predominant form in immature Sertoli cells. The effect of T(3) on its mRNA accumulation was quantified and confirmed by RT-PCR (control: 0.65 +/- 0.17; T(3): 1.23 +/- 0.04, arbitrary units, p < 0.05). However, transfection experiments showed that T(3) does not directly regulate GLUT1 promoter in any cell line tested. This is confirmed by the evidence that, upon extensive analysis, the rat GLUT1 promoter and the first intron sequence do not shows any thyroid responsive elements. Our data demonstrate that GLUT1 and GLUT8 are both expressed in prepubertal testis, but only GLUT1 is regulated by T(3). In addition, we found that the effect of T(3) cannot be attributed to its action on GLUT1 promoter.
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