HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
批准号:
3318743
负责人:
WILLIAM C BUHI
金额:
$10.34万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1986
资助国家:
美国
项目状态:
已结题
起止时间:
1986-04-01 至 1993-03-31
关键词:
antibody formation complementary DNA early embryonic stage egg /ovum estrogens estrus fallopian tubes fertilization gel electrophoresis gel filtration chromatography genetic library genetic manipulation genetic transcription genetic translation genital secretion glycoproteins histopathology hormone regulation /control mechanism humoral immunity immunochemistry immunosuppression laboratory rabbit macromolecule membrane proteins messenger RNA molecular cloning monoclonal antibody nucleic acid probes ovariectomy peptide hormone progesterone protein biosynthesis protein sequence recombinant DNA secretory protein sex hormones sperm swine
中文摘要
这项研究的长期目标是增加我们的知识
输卵管功能和了解复杂的生化
以及发生在精子、卵子和
植入前胚胎与母体输卵管及其分泌物
允许受精,早期卵裂阶段的胚胎
妊娠的发展和确立。这样做的目的是
研究建议是:1)完成鉴定,
De-nevo的表征、分布和纯化
合成输卵管分泌蛋白(OSP),特别是
雌激素诱导的酸性115,000和85,000 mR蛋白;2)
测试OSP的功能;3)研究OSP的结构和
重组DNA技术对OSP的调控作用我们会
全程定量生产从头合成的OSP
玛瑙类固醇处理母猪的发情周期、妊娠早期和
分别观察壶腹和峡部的贡献。
它们合成A4e的时间跨度。我们将净化
发情前期出现的主要OSP(115Kd、85Kd、Piless Than 4)
并通过离子交换、亲和等方法从外植体培养基中发情
和/或凝胶过滤层析。多克隆生物的生产
主要OSP的抗体将允许免疫细胞化学检测
在输卵管上皮、精子、未受精卵和早期
卵裂期的胚胎,可能暗示有功能。超微结构
分析和组织学改变将与OSP相关
综合与分配。使用体外测试,我们将检查
OSP是否具有免疫抑制活性。此外,我们还将
确定单独或组合的OSP是否具有补充性
抑制活性和调节母体体液免疫
系统。我们将确定精子、卵子或胚胎
特异性结合OSP或卵子是否与早期卵裂
胚胎通过内吞作用摄取OSP。将使用体外培养
以检验输卵管是否能刺激胚胎发育。
我们将提纯主要的OSP(115Kd和
85Kd),检测有无微粒体的mRNA翻译
并构建了主要基因的cdna文库。
包括编码115K和蛋白质的那些。特异性cDNAs
将被克隆和鉴定,推导出氨基酸序列
和蛋白质数据库进行身份验证。CDNAs将是
用于探针,以确定特定OSP nRNA的量,并
研究发情周期的调节或激素的作用
治疗。这些研究可以确定和定义机制和
受精和胚胎发育的控制。
英文摘要
The long term goals of this research are to increase our knowledge
of oviductal functions and to understand the complex biochemical
and molecular interactions that occur between sperm, ova, and the
preimplantation embryo and the maternal oviduct and its secretions
that allow fertilization, early cleavage-stage embryonic
development and establishment of pregnancy. The goal of this
research proposal is: 1) To complete the identification,
characterization, distribution and purification of de novo
synthesized oviductal secretory proteins (OSP), in particular the
estrogen-induced acidic 115,000 Mr and 85,000 Mr proteins; 2) To
test for function of OSP; and 3) To study the structure and
regulation of OSP by recombinant DNA techniques. We will
quantitate production of de novo synthesized OSP throughout the
estrous cycle, early pregnancy, in onyx steroid-treated gilts and
examine the contribution of the ampulla and isthmus in each and
the time span during which they a4e synthesized. We will purify
the major OSP (115Kd, 85Kd, pIless than 4) that appear at proestrus
and estrus from explant culture medium by ion-exchange, affinity
and/or gel filtration chromatography. Production of polyclonal
antibodies to the major OSP will allow innunocytochemical detection
on oviductal epithelium, sperm, unfertilized ova, and early
cleavage-stage embryos and may suggest function. Ultrastructural
analysis and histological changes will be correlated with OSP
synthesis and distribution. Using in vitro tests we will examine
whether OSP have immunosuppressive activity. Further, we will
determine whether individual or combined OSP have complement
inhibitory activity and can regulate the maternal humoral immune
system. We will determine whether sperm, ova, or embryos
specifically bind OSP or whether ova and early cleavage-stage
embryos take up OSP by endocytosis. In vitro cultures will be used
to examine whether the oviduct can stimulate embryonic development.
We will purify polyadenylated mRNA for the major OSP (115Kd and
85Kd), examine mRNA translation with and without microsomal
membranes, and construct a cDNA library of major mRNA species
including those encoding the 115K and proteins. Specific cDNA's
will be cloned and characterized, the amino acid sequences derived
and protein data base examined for identity. The cDNA's will be
used for probes to determine amounts of specific OSP nRNA and to
study regulation during the estrous cycle or with hormone
treatment. These studies may identify and define mechanisms and
controls for fertilization and embryonic development.
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Purification and characterization of a uterine retinol-binding protein in the bitch.
母犬子宫视黄醇结合蛋白的纯化和表征。
DOI:
10.1042/bj3110407
发表时间:
1995
期刊:
The Biochemical journal
影响因子:
--
作者:
[Buhi,WC, Alvarez,IM, Shille,VM, Thatcher,MJ, Harney,JP, Cotton,M]
通讯作者:
Cotton,M
Synthesis and secretion of proteins by postpartum human oviductal tissue in culture.
培养中的产后人类输卵管组织蛋白质的合成和分泌。
DOI:
10.1016/s0015-0282(16)60432-6
发表时间:
1989
期刊:
Fertility and sterility
影响因子:
6.7
作者:
[Buhi,WC, VanWert,JW, Alvarez,IM, Dones-Smith,MM, Bernhisel,MA]
通讯作者:
Bernhisel,MA
Identification and immunolocalization of proteins synthesized by dog endometrium and membranes.
狗子宫内膜和膜合成的蛋白质的鉴定和免疫定位。
DOI:
--
发表时间:
1993
期刊:
Journal of reproduction and fertility. Supplement
影响因子:
--
作者:
[Buhi,WC, Shille,VM, Thatcher,MJ, Alvarez,IM, Qiu,YX]
通讯作者:
Qiu,YX
Distribution of lectin binding sites in the oviducts of cycling and hormone-treated pigs.
循环和激素处理猪输卵管中凝集素结合位点的分布。
DOI:
10.1002/jez.1402650608
发表时间:
1993
期刊:
The Journal of experimental zoology
影响因子:
--
作者:
[Raychoudhury,SS, Suarez,SS, Buhi,WC]
通讯作者:
Buhi,WC
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
-
批准号:3448231
-
项目类别:
-
资助金额:$4.49万
-
财政年份:1986
-
负责人:WILLIAM C BUHI
-
依托单位:
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
-
批准号:3448230
-
项目类别:
-
资助金额:$4.67万
-
财政年份:1986
-
负责人:WILLIAM C BUHI
-
依托单位:
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
-
批准号:3318739
-
项目类别:
-
资助金额:$10.77万
-
财政年份:1986
-
负责人:WILLIAM C BUHI
-
依托单位:
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
-
批准号:3448229
-
项目类别:
-
资助金额:$5.01万
-
财政年份:1986
-
负责人:WILLIAM C BUHI
-
依托单位:
HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS
-
批准号:3318742
-
项目类别:
-
资助金额:$9.95万
-
财政年份:1986
-
负责人:WILLIAM C BUHI
-
依托单位:
海外基金