Mayerhofer D, Mayerhofer A, Bartke A
Department of Physiology, School of Medicine, Southern Illinois University, Carbondale 62901-6512.
Int J Androl. 1992 Jun;15(3):263-81. doi: 10.1111/j.1365-2605.1992.tb01347.x.
We have developed an isolation and cultivation protocol for testicular macrophages from the seasonally breeding Siberian hamster, Phodopus sungorus which allows serum-free culture. Macrophages were isolated from the active testes of adult Phodopus kept in long photoperiod (LD, 18hL:6hD), and from the inactive testes of animals with gonadal regression induced by exposure to a short photoperiod (SD, 6hL;18hD; for 10 weeks). The isolated cells were identified as macrophages by (a) electron microscopy, (b) non-specific phagocytosis, and (c) the presence of Fc-receptors. Treatment of the cultures with oFSH (at 0.5 and 1 micrograms ml-1 for 1 and 4 days) significantly stimulated lactate secretion by testicular macrophages from LD hamsters, as expected from studies in the rat. In marked contrast, oFSH did not affect lactate secretion by testicular macrophages from SD hamsters, or by peritoneal macrophages. To test possible macrophage-Leydig cell interactions, the influence of testicular macrophage-conditioned medium (TM-CM) from FSH-responsive LD macrophage cultures was assessed and compared with the effect of peritoneal macrophage-conditioned medium (PM-CM) on testosterone production by testicular parenchyma from LD hamsters. Unexpectedly, and in contrast to previous reports in rats, testosterone production by testes was inhibited significantly by TM-CM from cultures pretreated with either 0.1 or 0.5 micrograms FSH ml-1. FSH alone stimulated testosterone production (due either to LH contamination or to possible paracrine effects). PM-CM from cultures pretreated with FSH did have an inhibitory effect. It is concluded that: (i) There is indirect evidence for the presence of functional FSH receptors on macrophages from the active testes of Phodopus. (ii) Testicular macrophages from inactive testes of Phodopus are not responsive to FSH, implicating impaired macrophage function in the regressed testes of Phodopus exposed to SD. (iii) Testicular macrophages produce a factor which can inhibit testosterone production by incubated testicular tissue.
我们已经开发出一种从季节性繁殖的西伯利亚仓鼠（Phodopus sungorus）中分离和培养睾丸巨噬细胞的方案，该方案允许无血清培养。巨噬细胞从处于长光照周期（LD，18小时光照：6小时黑暗）的成年Phodopus的活跃睾丸中分离出来，以及从暴露于短光照周期（SD，6小时光照；18小时黑暗，持续10周）诱导性腺退化的动物的不活跃睾丸中分离出来。通过以下方法将分离出的细胞鉴定为巨噬细胞：（a）电子显微镜检查，（b）非特异性吞噬作用，以及（c）Fc受体的存在。如在大鼠研究中所预期的那样，用oFSH（浓度为0.5和1微克/毫升，处理1天和4天）处理培养物显著刺激了来自LD仓鼠的睾丸巨噬细胞的乳酸分泌。与之形成鲜明对比的是，oFSH对来自SD仓鼠的睾丸巨噬细胞或腹膜巨噬细胞的乳酸分泌没有影响。为了测试巨噬细胞与睾丸间质细胞之间可能的相互作用，评估了来自FSH反应性LD巨噬细胞培养物的睾丸巨噬细胞条件培养基（TM-CM）的影响，并与腹膜巨噬细胞条件培养基（PM-CM）对来自LD仓鼠的睾丸实质睾酮产生的影响进行了比较。出乎意料的是，与之前大鼠的报道相反，用0.1或0.5微克FSH/毫升预处理的培养物中的TM-CM显著抑制了睾丸的睾酮产生。单独的FSH刺激了睾酮产生（这可能是由于LH污染或可能的旁分泌作用）。用FSH预处理的培养物中的PM-CM确实有抑制作用。得出以下结论：（i）有间接证据表明来自Phodopus活跃睾丸的巨噬细胞上存在功能性FSH受体。（ii）来自Phodopus不活跃睾丸的睾丸巨噬细胞对FSH无反应，这意味着暴露于SD的Phodopus退化睾丸中的巨噬细胞功能受损。（iii）睾丸巨噬细胞产生一种可抑制孵育的睾丸组织睾酮产生的因子。
