DeSouza M M, Murray M K
Department of Anatomy and Cellular Biology, Tufts University, School of Veterinary Medicine, Boston, Massachusetts 02111, USA.
Endocrinology. 1995 Jun;136(6):2485-96. doi: 10.1210/endo.136.6.7750470.
An estrogen (E2)-dependent 90,000-92,000 M(r) protein is synthesized and released by the sheep oviduct in a temporally and regionally specific manner during the first few days of pregnancy, where it associates with the embryo. The present study was undertaken to further define the nature of this protein and its regulation at a time when fertilization and early embryonic cleavage occur in the oviduct. The complete complementary DNA (cDNA) sequence for the 90,000-92,000 M(r) protein was determined, and steady state messenger RNA (mRNA) levels were measured during early pregnancy (estrus and days 1, 2, 3, 4, 6, and 16). The composite cDNA possessed an open reading frame of 1633 bases with a single potential N-glycosylation site. The inferred amino acid (aa) sequence predicted a prepolypeptide of 539 aa (69,151 M(r)) and a mature polypeptide of 518 aa (66,477 M(r)). Nucleotide and deduced aa sequence shared identity with translated E2-dependent cow, human, and partially sequenced baboon oviduct protein cDNAs; a human articular cartilage protein, gp 39; and chitinases. Northern blot hybridization revealed a single RNA species (2.2 kilobases) in the fimbria and ampulla, which was not detected in the isthmus or other reproductive and nonreproductive tract tissue RNAs. Steady state levels of mRNA encoding the 90,000-92,000 M(r) were highest in the fimbria and ampulla at estrus and on day 1 of pregnancy, when gamete transport and fertilization occur in the E2-dominated fallopian tube. Oviduct protein mRNA levels declined significantly (P < 0.05) on day 2 and underwent a further significant reduction on day 3 of pregnancy coincident with transport of the embryo from the oviduct to the uterus, a reproductive stage associated with rising progesterone levels. By day 16 of pregnancy, the mRNA encoding the E2-dependent oviduct protein was virtually undetectable. The temporally and regionally specific regulation of gene expression for the 90,000-92,000 M(r) E2-dependent protein correlates with protein synthesis data and emphasizes the precise regulation of its synthesis at a time when fertilization and embryonic cleavage are taking place in the oviduct. Identity with a growing class of steroid-regulated oviduct secretory proteins and chitinase enzymes suggests a possible structural and/or functional relationship, which may be important in mediating these latter reproductive events.
在怀孕的头几天,绵羊输卵管以时间和区域特异性的方式合成并释放一种雌激素(E2)依赖性的90,000 - 92,000 M(r)蛋白,该蛋白与胚胎相关联。本研究旨在进一步确定这种蛋白的性质及其在输卵管发生受精和早期胚胎分裂时的调控机制。测定了90,000 - 92,000 M(r)蛋白的完整互补DNA(cDNA)序列,并在怀孕早期(发情期以及第1、2、3、4、6和16天)测量了稳态信使RNA(mRNA)水平。复合cDNA拥有一个1633个碱基的开放阅读框,带有一个潜在的N - 糖基化位点。推断的氨基酸(aa)序列预测出一个539个aa(69,151 M(r))的前体多肽和一个518个aa(66,477 M(r))的成熟多肽。核苷酸和推导的aa序列与已翻译的E2依赖性牛、人类以及部分测序的狒狒输卵管蛋白cDNA、一种人类关节软骨蛋白gp 39和几丁质酶具有同源性。Northern印迹杂交显示在伞部和壶腹部有单一的RNA种类(2.2千碱基),在峡部或其他生殖和非生殖管道组织RNA中未检测到。编码90,000 - 92,000 M(r)蛋白的mRNA稳态水平在发情期和怀孕第1天的伞部和壶腹部最高,此时配子运输和受精发生在以E2为主的输卵管中。输卵管蛋白mRNA水平在怀孕第2天显著下降(P < 0.05),并在怀孕第3天随着胚胎从输卵管向子宫的运输进一步显著降低,这一生殖阶段与孕酮水平升高相关。到怀孕第16天,编码E2依赖性输卵管蛋白的mRNA几乎检测不到。90,000 - 92,000 M(r)E2依赖性蛋白基因表达的时间和区域特异性调控与蛋白质合成数据相关,并强调了在输卵管发生受精和胚胎分裂时其合成的精确调控。与越来越多的类固醇调节的输卵管分泌蛋白和几丁质酶的同源性表明可能存在结构和/或功能关系,这可能在介导这些后期生殖事件中很重要。