GENE EXPRESSION IN EMBRYOS CULTURED IN VITRO

体外培养胚胎中的基因表达

基本信息

批准号：
2889312
负责人：
MARK E WESTHUSIN
金额：
$ 26.22万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-09-01 至 2000-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2889312
关键词：
confocal scanning microscopy cooperative study cow developmental genetics early embryonic stage embryo /fetus tissue /cell culture embryogenesis gene expression growth factor receptors immunofluorescence technique in vitro fertilization insulinlike growth factor interferons messenger RNA method development polymerase chain reaction secretion sheep

项目摘要

DESCRIPTION (Adapted from applicant's description): Less than 10% of transferred human IVF embryos establish a pregnancy. In other species, the losses are not as high, but are appreciable. Some in vitro embryo culture systems are associated abnormal phenotypes including large birth weights and often embryonic, fetal or newborn death. Embryos that result in abnormally large offspring display a normal morphology when transferred. This highlights the importance of evaluating culture influences on genetic characteristics in addition to morphological parameters. Culture environments must not only sustain development and morphology, but also maintain normal patterns of gene expression. The overall goal of this proposal is to develop safe, effective, and efficient methods for mammalian embryo culture. The specific aims will evaluate the quality of bovine embryos produced by a variety of in vitro culture methods by determining the influence of culture on insulin-like growth factor (IGF) and interferon tau (bTP1) gene expression. These molecules are important targets for examining the unusual growth rates of cultured bovine embryos as altered levels of these genes are likely to reflect changes in the developmental program of early embryogenesis. The experiments will examine the following specific aims: 1: To determine the effects of culture on the expression of mRNAs encoding IGFI, IGFII and their receptors. 2: To determine the effect of culture on the cellular distribution of IGF polypeptides. 3: To determine the effects of culture on expression and secretion of bTP1. 4: To determine the influence of culture on IGF secretion. 5: To determine the effect of culture on bovine embryo development. 6: To isolate and characterize culture sensitive differentially expressed transcripts (mRNAs) in early embryos using differential display techniques (RT-DDPLR).

描述（根据申请人的描述改编）：不到10％转移的人IVF胚胎建立了妊娠。在其他物种中损失不那么高，但很明显。一些体外胚胎培养系统是相关的异常表型，包括大的出生体重和经常胚胎，胎儿或新生儿死亡。导致异常的胚胎转移时，大后代显示出正常的形态。这强调评估文化对遗传的影响的重要性除形态参数外。文化环境不仅必须维持发展和形态，还必须保持基因表达的正常模式。总体目标建议是为哺乳动物开发安全，有效和有效的方法胚胎文化。具体目的将评估牛的质量通过确定多种体外培养方法产生的胚胎培养对胰岛素样生长因子（IGF）和干扰素TAU的影响（BTP1）基因表达。这些分子是检查的重要目标培养的牛胚胎的异常增长速率随着水平的变化这些基因可能反映了发展计划的变化早期胚胎发生。实验将检查以下特定目的：1：确定培养对mRNA表达的影响编码IGFI，IGFII及其受体。 2：确定培养IGF多肽的细胞分布。 3：确定培养对BTP1表达和分泌的影响。 4：到确定培养对IGF分泌的影响。 5：确定培养对牛胚胎发育的影响。 6：孤立和表征培养敏感差异表达的转录本（mRNA）在早期的胚胎中，使用微分显示技术（RT-DDPLR）。