Kitahara S, Winters S J, Attardi B, Oshima H, Troen P
Department of Medicine, Montefiore Hospital, Pittsburgh, Pennsylvania 15213.
Endocrinology. 1990 May;126(5):2642-9. doi: 10.1210/endo-126-5-2642.
Because the role of the pituitary in the testicular control of gonadotropin secretion remains controversial, we examined the effects of castration on the release of LH and FSH under basal conditions and in response to GnRH stimulation by dispersed pituitary cells in monolayer culture as well as by cells perifused with pulses of GnRH. These effects were compared to changes in LH beta, FHS beta, and alpha-subunit mRNA levels determined by Northern blot analysis. Pituitary cells were prepared from 7-week-old intact rats and rats orchidectomized 2 weeks previously. Castration increased basal FSH secretion from monolayer cultures, interpulse FSH release from perifused pituitary cells, FSH beta mRNA concentrations and serum FSH levels each approximately 2-fold, whereas pituitary FSH contents were similar in cells from intact and castrated rats. Pituitary LH content rose 3-fold, LH beta mRNA rose 5.6-fold, and basal LH secretion increased 6-fold, but serum LH levels increased 22-fold. Thus, the change in FSH synthesis inferred from the increase in FSH beta mRNA was proportional to the increase in FSH secretion both in vitro and in vivo. Whereas the basal release of LH in vitro was also proportional to the change in LH beta mRNA, secretion of LH in vivo exceeded these changes, underscoring the importance of increased GnRH to the serum LH castration response. Castration resulted in an increase in the sum of FSH content and secretion during 10 days in culture in the absence of GnRH, indicating ongoing FSH synthesis. Total LH declined in cells from intact rats, and this decline was prevented by castration; this effect may be due to a castration-related decrease in intracellular LH degradation or increased LH synthesis in the absence of GnRH. Castration also augmented the GnRH-stimulated release of LH and FSH from monolayer cultures 4.5- and 1.8-fold, respectively, and increased the amplitude of GnRH-stimulated LH and FSH pulses 5- and 2-fold in experiments with perifused pituitary cells. The EC50 for GnRH was unaffected by castration.(ABSTRACT TRUNCATED AT 400 WORDS)
由于垂体在睾丸对促性腺激素分泌的调控中所起的作用仍存在争议,我们研究了阉割对基础条件下以及单层培养的分散垂体细胞和经GnRH脉冲灌流的细胞在GnRH刺激下LH和FSH释放的影响。这些影响与通过Northern印迹分析测定的LHβ、FSHβ和α亚基mRNA水平的变化进行了比较。垂体细胞取自7周龄的完整大鼠以及2周前进行了睾丸切除的大鼠。阉割使单层培养物中的基础FSH分泌、灌流垂体细胞的脉冲间期FSH释放、FSHβ mRNA浓度和血清FSH水平各自增加了约2倍,而完整大鼠和阉割大鼠的垂体FSH含量相似。垂体LH含量增加了3倍,LHβ mRNA增加了5.6倍,基础LH分泌增加了6倍,但血清LH水平增加了22倍。因此,从FSHβ mRNA增加推断出的FSH合成变化在体外和体内均与FSH分泌的增加成比例。虽然体外LH的基础释放也与LHβ mRNA的变化成比例,但体内LH的分泌超过了这些变化,突出了GnRH增加对血清LH阉割反应的重要性。阉割导致在无GnRH的情况下培养10天期间FSH含量和分泌的总和增加,表明FSH持续合成。完整大鼠细胞中的总LH下降,而阉割可防止这种下降;这种效应可能是由于阉割相关的细胞内LH降解减少或在无GnRH时LH合成增加。阉割还分别使单层培养物中GnRH刺激的LH和FSH释放增加了4.5倍和1.8倍,并在垂体细胞灌流实验中使GnRH刺激的LH和FSH脉冲幅度分别增加了5倍和2倍。GnRH的半数有效浓度不受阉割影响。(摘要截短至400字)