Higgins S J, Young P, Brody J R, Cunha G R
Department of Biochemistry, University of Leeds, UK.
Development. 1989 Jun;106(2):219-34. doi: 10.1242/dev.106.2.219.
Functional cytodifferentiation of seminal vesicle epithelium was investigated in tissue recombinants. Neonatal rat and mouse seminal vesicles were separated into epithelium and mesenchyme using trypsin. Epithelium and mesenchyme were then recombined in vitro to form interspecific rat/mouse homotypic recombinants. Growth as renal grafts in adult male athymic mice resulted in seminal vesicle morphogenesis in 70% of the recombinants (the remaining 30% failed to grow). Functional cytodifferentiation was judged by the expression of the major androgen-dependent secretory proteins characteristic of the seminal vesicles of adult rats and mice. Antibodies specific for each of these proteins were used to screen tissue sections by immunocytochemistry and to probe protein extracts by immunoblotting techniques. The heterospecific recombinants synthesized the full range of seminal vesicle secretory proteins that typifies the species providing the epithelium of the recombinant, not the mesenchyme. There was little functional variation between individual recombinants. The time course of development corresponded to that of intact neonatal seminal vesicles grown under the same conditions. Morphogenesis and functional cytodifferentiation were not evident after one week, but were well advanced after two weeks. Seminal vesicle recombinants grown for three weeks were indistinguishable morphologically and functionally from normal adult seminal vesicles. In addition, the ability of adult seminal vesicle epithelium to be induced to proliferate was examined. In association with neonatal seminal vesicle mesenchyme, the epithelium of the adult seminal vesicle proliferated and retained its normal functional activity. Thus, seminal vesicle functional cytodifferentiation can be faithfully reproduced in homotypic tissue recombinants. The methods used in this study will be used to investigate seminal vesicle development in instructive inductions of heterotypic epithelia.
在组织重组体中研究了精囊上皮的功能性细胞分化。使用胰蛋白酶将新生大鼠和小鼠的精囊分离为上皮和间充质。然后将上皮和间充质在体外重组以形成种间大鼠/小鼠同型重组体。在成年雄性无胸腺小鼠中作为肾移植物生长导致70%的重组体发生精囊形态发生(其余30%未能生长)。通过成年大鼠和小鼠精囊中主要雄激素依赖性分泌蛋白的表达来判断功能性细胞分化。针对这些蛋白质中的每一种的特异性抗体用于通过免疫细胞化学筛选组织切片并通过免疫印迹技术探测蛋白质提取物。异种特异性重组体合成了代表提供重组体上皮而非间充质的物种的全套精囊分泌蛋白。各个重组体之间几乎没有功能差异。发育的时间进程与在相同条件下生长的完整新生精囊的时间进程一致。一周后形态发生和功能性细胞分化不明显,但两周后进展良好。生长三周的精囊重组体在形态和功能上与正常成年精囊无法区分。此外,还研究了成年精囊上皮被诱导增殖的能力。与新生精囊间充质相关,成年精囊上皮增殖并保留其正常功能活性。因此,精囊功能性细胞分化可以在同型组织重组体中如实地再现。本研究中使用的方法将用于研究异型上皮的诱导性诱导中精囊的发育。