IBMX和激素对大鼠支持细胞基因表达的影响

Indrashis Bhattacharya , Mukkesh Gautam , Subeer S. Majumdar
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引用次数: 1

摘要

支持细胞(Sc)在卵泡刺激素(FSH)和睾酮(T)的控制下调节精子发生。Sc在青春期发育过程中支持精子发生的功能成熟是男性生育能力的先决条件。然而,激素驱动Sc成熟变化的作用尚不清楚。在本研究中,我们比较了从新生大鼠(9日龄)和青春期前大鼠(18日龄)睾丸中分离的未成熟Sc和成熟Sc的激素诱导基因表达,以探讨Sc在出生后成熟过程中激素反应性的发育差异以及3-异丁基-1-甲基黄嘌呤(IBMX)的影响。结果和结论我们的研究结果表明,与9日龄的Sc相比,18日龄大鼠获得的Sc在乳酸、cAMP和基因转录方面的FSH反应性更为突出。我们的结果还表明,尽管在Sc原代培养中使用IBMX在FSH诱导的cAMP反应方面产生了更好的结果,但这种药理学剂的存在缓和了FSH刺激的基因表达谱。我们的数据进一步表明,如果持续补充FSH和T(联合)培养,未成熟的Sc能够在体外分化。综上所述,我们还得出结论,为了准确评估激素对基因表达的调节作用,应避免在Sc原代培养中使用IBMX。
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The effect of IBMX and hormones on gene expression by rat Sertoli cells

Background

Sertoli cells (Sc) regulate spermatogenesis under the control of FSH and testosterone (T). Functional maturation of Sc for supporting the spermatogenic onset during pubertal development is prerequisite for male fertility. However, the effect of hormone driven maturational changes in Sc is not well known.

Objectives and experimental model

In this present study we have compared hormone induced gene expression of immature and mature Sc isolated from neonatal (9-days old) and prepubertal (18-days-old) rat testes, respectively, to investigate the developmental difference of hormone responsiveness of Sc during postnatal maturation as well as influence of 3-isobutyl-1-methylxanthine (IBMX), a nonspecific inhibitor of phosphodiesterase in primary culture of Sc.

Results and conclusion

Our results suggested that FSH responsiveness of Sc obtained from 18-days-old rats were more prominent in terms of augmentation of lactate, cAMP and gene transcription as compared to Sc from 9-days of age. Our result also indicated that although the use of IBMX in primary culture of Sc generates a better readout in terms of FSH induced cAMP response, the presence of such pharmacological agent mellows down FSH stimulated gene expression profile. Our data indicated further that immature Sc are capable of differentiating in vitro if cultured with continuous supplementation of FSH and T (in combination). Taken together, we also concluded that for accurate evaluation of the modulation of gene expression by hormones, use of IBMX should be avoided in primary cultures of Sc.

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