Amelkina Olga, Zschockelt Lina, Painer Johanna, Serra Rodrigo, Villaespesa Francisco, Braun Beate C, Jewgenow Katarina
Department of Reproduction Biology, Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany.
Iberian lynx captive breeding program, Centro Nacional de Reprodução de Lince Ibérico, Silves, Portugal.
PLoS One. 2015 Nov 24;10(11):e0143414. doi: 10.1371/journal.pone.0143414. eCollection 2015.
The corpus luteum (CL) is a transient gland formed in the ovary after ovulation and is the major source of progesterone. In the Iberian and Eurasian lynx, CL physiologically persist after parturition and retain their capacity to produce progesterone, thus suppressing the ovarian activity. This unique reproductive characteristic has a big impact on the success of assisted reproduction techniques in the endangered Iberian lynx. The mechanisms behind CL persistence are not yet understood and require extensive studies on potential luteotropic and luteolytic factors in felids. Because the apoptosis system has been shown to be involved in structural regression of CL in many species, we aimed to investigate the capacity of perCL to undergo apoptosis. In addition, we performed initial studies on the apoptosis system in the luteal phase of the domestic cat. No previous research on this system has been made in this species. Our factors of interest included agents of the intrinsic apoptosis pathway, i.e., pro-survival B-cell CLL/lymphoma 2 (BCL2) and pro-apoptotic BCL2-associated X protein (BAX), the executioner caspase-3 (CASP3), as well as of the extrinsic pathway, i.e., pro-apoptotic receptor FAS, and tumor necrosis factor (TNF) and its receptors (pro-apoptotic TNFRSF1A and pro-survival TNFRSF1B). We analyzed the relative mRNA levels of these factors, as well as protein localization of CASP3 and TNF during stages of pregnancy and the non-pregnant luteal phase in CL of domestic cats. The same factors were investigated in freshly ovulated CL (frCL) and perCL of Iberian and Eurasian lynx, which were histologically analyzed. All factors were present in the CL tissue of both domestic cat and lynx throughout all analyzed stages. The presence of pro-apoptotic factors BAX, CASP3, FAS and TNFRSF1A in perCL of the Eurasian and Iberian lynx might indicate the potential sensitivity of perCL to apoptotic signals. The expression of pro-survival factors BCL2 and TNFRSF1B was significantly higher in perCL compared to frCL of studied Iberian lynx, suggesting the potential involvement of these factors in the structural integrity of perCL. In both Iberian lynx and pregnant and non-pregnant domestic cats, the expression of TNFRSF1A was significantly higher in forming CL compared to other stages, suggesting the conserved involvement of this factor in the tissue reorganization during formation of the feline CL. The mRNA levels of CASP3 and TNFRSF1B were highest during regression stages of domestic cat CL. The current study provides initial results on the possible involvement of the apoptosis system in the structure and function of the feline CL and in its physiological persistence.
黄体(CL)是排卵后在卵巢中形成的临时性腺体,是孕酮的主要来源。在伊比利亚猞猁和欧亚猞猁中,黄体在分娩后仍会生理性持续存在,并保留产生孕酮的能力,从而抑制卵巢活动。这种独特的生殖特征对濒危的伊比利亚猞猁辅助生殖技术的成功有着重大影响。黄体持续存在背后的机制尚不清楚,需要对猫科动物中潜在的促黄体生成和黄体溶解因子进行广泛研究。由于凋亡系统已被证明在许多物种的黄体结构退化中起作用,我们旨在研究产后黄体(perCL)发生凋亡的能力。此外,我们对家猫黄体期的凋亡系统进行了初步研究。此前尚未对该物种的这个系统进行过研究。我们感兴趣的因子包括内源性凋亡途径的因子,即抗凋亡的B细胞淋巴瘤/白血病-2(BCL2)和促凋亡的BCL2相关X蛋白(BAX)、执行凋亡的半胱天冬酶-3(CASP3),以及外源性途径的因子,即促凋亡受体FAS、肿瘤坏死因子(TNF)及其受体(促凋亡的TNFRSF1A和抗凋亡的TNFRSF1B)。我们分析了这些因子的相对mRNA水平,以及家猫黄体在妊娠阶段和非妊娠黄体期CASP3和TNF的蛋白定位。在伊比利亚猞猁和欧亚猞猁的新鲜排卵黄体(frCL)和perCL中研究了相同的因子,并进行了组织学分析。在所有分析阶段,家猫和猞猁的黄体组织中均存在所有这些因子。欧亚猞猁和伊比利亚猞猁perCL中促凋亡因子BAX、CASP3、FAS和TNFRSF1A的存在可能表明perCL对凋亡信号具有潜在敏感性。与所研究的伊比利亚猞猁的frCL相比,perCL中抗凋亡因子BCL2和TNFRSF1B的表达显著更高,表明这些因子可能参与了perCL的结构完整性维持。在伊比利亚猞猁以及妊娠和非妊娠的家猫中,与其他阶段相比,TNFRSF1A在黄体形成过程中的表达显著更高,表明该因子在猫科动物黄体形成过程中的组织重组中持续发挥作用。家猫黄体退化阶段CASP3和TNFRSF1B的mRNA水平最高。本研究为凋亡系统可能参与猫科动物黄体的结构和功能及其生理持续性提供了初步结果。