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猫科动物类固醇激素代谢与卵巢活动的比较研究,通过粪便进行非侵入性测量。

Comparative aspects of steroid hormone metabolism and ovarian activity in felids, measured noninvasively in feces.

作者信息

Brown J L, Wasser S K, Wildt D E, Graham L H

机构信息

Conservation and Research Center, Smithsonian Institution, Front Royal, Virginia 22630.

出版信息

Biol Reprod. 1994 Oct;51(4):776-86. doi: 10.1095/biolreprod51.4.776.

Abstract

Noninvasive fecal assays were used to study steroid metabolism and ovarian activity in several felid species. Using the domestic cat (Felis catus) as model, the excretory products of injected [14C]estradiol (E2) and [14C]progesterone (P4) were determined. Within 2 days, 97.0 +/- 0.6% and 96.7 +/- 0.5% of recovered E2 and P4 radioactivity, respectively, was found in feces. E2 was excreted as unconjugated estradiol and estrone (40%) and as a non-enzyme-hydrolyzable conjugate (60%). P4 was excreted primarily as non-enzyme-hydrolyzable, conjugated metabolites (78%) and as unconjugated pregnenolone epimers. A simple method for extracting fecal steroid metabolites optimized extraction efficiencies of the E2 and P4 excretion products (90.1 +/- 0.8% and 87.2 +/- 1.4%, respectively). Analysis of HPLC fractions of extracted fecal samples from the radiolabel-injected domestic cats revealed that E2 immunoreactivity coincided primarily with the unconjugated metabolized [14C]E2 peak, whereas progestogen immunoreactivity coincided with a single conjugated epimer and multiple unconjugated pregnenolone epimers. After HPLC separation, similar immunoreactive E2 and P4 metabolite profiles were observed in the leopard cat (F. bengalensis), cheetah (Acinonyx jubatus), clouded leopard (Neofelis nebulosa), and snow leopard (Panthera uncia). Longitudinal analyses demonstrated that changes in fecal E2 and P4 metabolite concentrations reflected natural or artificially induced ovarian activity. For example, severalfold increases in E2 excretion were associated with overt estrus or exogenous gonadotropin treatment, and elevated fecal P4 metabolite concentrations occurred during pregnant and nonpregnant (pseudopregnant) luteal phases. Although overall concentrations were similar, the duration of elevated fecal P4 metabolites during pseudopregnancy was approximately half that observed during pregnancy. In summary, steroid metabolism mechanisms appear to be conserved among these physically diverse, taxonomically related species. Results indicate that this hormone-monitoring approach will be extremely useful for elucidating the hormonal regulatory mechanism associated with the reproductive cycle, pregnancy, and parturition of intractable and endangered felid species.

摘要

采用非侵入性粪便检测方法研究了几种猫科动物的类固醇代谢和卵巢活性。以家猫(Felis catus)为模型,测定了注射[14C]雌二醇(E2)和[14C]孕酮(P4)后的排泄产物。在2天内,粪便中分别发现回收的E2和P4放射性的97.0±0.6%和96.7±0.5%。E2以未结合的雌二醇和雌酮(40%)以及一种非酶可水解结合物(60%)的形式排泄。P4主要以非酶可水解的结合代谢物(78%)和未结合的孕烯醇酮差向异构体的形式排泄。一种简单的粪便类固醇代谢物提取方法优化了E2和P4排泄产物的提取效率(分别为90.1±0.8%和87.2±1.4%)。对注射放射性标记的家猫提取粪便样本的高效液相色谱(HPLC)馏分分析表明,E2免疫反应性主要与未结合的代谢[14C]E2峰一致,而孕激素免疫反应性与单一结合差向异构体和多种未结合的孕烯醇酮差向异构体一致。HPLC分离后,豹猫(F. bengalensis)、猎豹(Acinonyx jubatus)、云豹(Neofelis nebulosa)和雪豹(Panthera uncia)中观察到类似的E2和P4免疫反应性代谢物谱。纵向分析表明,粪便E2和P4代谢物浓度的变化反映了自然或人工诱导的卵巢活性。例如,E2排泄量增加几倍与明显发情或外源性促性腺激素治疗有关,粪便P4代谢物浓度升高发生在怀孕和非怀孕(假孕)黄体期。虽然总体浓度相似,但假孕期间粪便P4代谢物升高的持续时间约为怀孕期间观察到的一半。总之,类固醇代谢机制在这些形态各异、分类学上相关的物种中似乎是保守的。结果表明,这种激素监测方法对于阐明与难处理和濒危猫科动物的生殖周期、怀孕和分娩相关的激素调节机制将极为有用。

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