Morena A R, Boitani C, de Grossi S, Stefanini M, Conti M
Institute of Histology and General Embryology, University of Rome, La Sapienza, Italy.
Endocrinology. 1995 Feb;136(2):687-95. doi: 10.1210/endo.136.2.7835302.
Four genes (ratPDE1/IVc, ratPDE2/IVa, ratPDE3/IVd, and ratPDE4/IVb) encoding different isoforms of phosphodiesterase that specifically hydrolyze the second messenger cAMP (cAMP-PDEs) are present in the rat. Previous data from our laboratory indicated that these genes are differentially expressed in the somatic and germ cells of the seminiferous epithelium of the testis. To further characterize their spatial and temporal expression in the seminiferous tubules, in situ hybridization was used to monitor the expression of the four cAMP-PDE messenger RNAs (mRNAs). The signals corresponding to ratPDE1/IVc and ratPDE2IVa mRNAs were localized in two restricted layers of the seminiferous epithelium. The ratPDE1/IVc mRNA was present in a region of the epithelium corresponding to the location of middle-late pachytene spermatocytes. Conversely, the ratPDE2/IVa signal was confined to a more adluminal area corresponding to the location of maturing round spermatids. The ratPDE3/IVd and ratPDE4/IVb mRNAs were distributed throughout the span of the seminiferous epithelium, indicating a localization in the Sertoli cell cytoplasm. Although the intensity of the signal corresponding to ratPDE4/IVb was similar in all seminiferous tubule stages, the ratPDE3/IVd signal varied in intensity in tubules at different stages of the seminiferous cycle. Maximal expression was present in tubules at stages I-V and XI-XIII of the cycle and minimal at stages VIII-IX of the cycle. The expression of the ratPDE3/IVd mRNA positively correlated with the ability of specific inhibitors of the cAMP-PDEs to potentiate the FSH-dependent cAMP accumulation in tubules at different stages of the seminiferous cycle, with maximal potentiation observed at stages II-VI of the cycle. These data demonstrate that different cAMP-PDE genes are active in different cells of the seminiferous tubules and that the ratPDE3/IVd gene is expressed in the Sertoli cell in a cyclical fashion during the seminiferous cycle.
大鼠体内存在四种编码磷酸二酯酶不同亚型的基因(大鼠磷酸二酯酶1/IVc、大鼠磷酸二酯酶2/IVa、大鼠磷酸二酯酶3/IVd和大鼠磷酸二酯酶4/IVb),这些磷酸二酯酶能特异性水解第二信使环磷酸腺苷(cAMP磷酸二酯酶)。我们实验室之前的数据表明,这些基因在睾丸生精上皮的体细胞和生殖细胞中存在差异表达。为了进一步明确它们在生精小管中的时空表达特征,采用原位杂交技术监测四种cAMP磷酸二酯酶信使核糖核酸(mRNA)的表达。与大鼠磷酸二酯酶1/IVc和大鼠磷酸二酯酶2/IVa mRNA对应的信号定位于生精上皮的两个特定层。大鼠磷酸二酯酶1/IVc mRNA存在于上皮细胞中对应于中晚期粗线期精母细胞位置的区域。相反,大鼠磷酸二酯酶2/IVa信号局限于更靠近管腔的区域,对应于成熟圆形精子细胞的位置。大鼠磷酸二酯酶3/IVd和大鼠磷酸二酯酶4/IVb mRNA分布于整个生精上皮,表明定位于支持细胞的细胞质中。尽管在所有生精小管阶段,与大鼠磷酸二酯酶4/IVb对应的信号强度相似,但大鼠磷酸二酯酶3/IVd信号在生精周期不同阶段的小管中强度有所变化。在周期的I-V期和XI-XIII期小管中表达最高,在周期的VIII-IX期最低。大鼠磷酸二酯酶3/IVd mRNA的表达与cAMP磷酸二酯酶特异性抑制剂在生精周期不同阶段增强小管中促卵泡激素(FSH)依赖性cAMP积累的能力呈正相关,在周期的II-VI期观察到最大增强作用。这些数据表明,不同的cAMP磷酸二酯酶基因在生精小管的不同细胞中具有活性,并且大鼠磷酸二酯酶3/IVd基因在生精周期中以周期性方式在支持细胞中表达。