BDNF in adult mouse taste cells

成年小鼠味觉细胞中的 BDNF

• 批准号: 6445946
• 负责人: CINDY L YEE
• 金额: $ 2.17万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-10 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6445946
• 关键词: brain derived neurotrophic factor; bromodeoxyuridine; cell biology; cell growth regulation; cell population study; cell type; denervation; fiber cell; ganglion cell; gene expression; gene targeting; genetically modified animals; growth /development; immunocytochemistry; innervation; laboratory mouse; mature animal; nerve; nervous system regeneration; neurosurgery; predoctoral investigator; sensory receptors; synapses; taste buds; tongue

The goals of this study are to use BDNF-LacZ transgenic "knock-in" mice to determine the role of BDNF in adult mouse taste cells. The first aim will determine which taste cell types express BDNF in the adult mouse. Specifically, whether BDNF is present in newly divided taste cells, immature taste cells, or phenotypically mature taste cells. If BDNF is present in mature taste cells, I am interested in determining which immunohistochemical and morphological classes of taste cells BDNF is present in. I will use double-label immunohistochemistry to achieve this aim. To birthdate cells I will inject mice with 5'-Bromo-2' deoxyuridine which incorporates into the DNA of dividing cells. I am also interested in determining what effect(s) loss of taste bud innervation will have on the expression of BDNF and what effect(s) subsequent regeneration and innervation of taste buds will have on the expression of BDNF. To achieve this aim I will surgically crush the gustatory nerves in BDNF-LacZ mice and let the mice live for successively longer days post nerve crush to look at the effect(s) nerve degeneration and regeneration has on BDNF expression in taste buds. For my final aim, 1 am interested in determining what effect(s) adult loss of BDNF expression in taste buds has on taste buds, gustatory nerve fibers and gustatory ganglion cells. Since I am interested in looking at the effect loss of BDNF expression has on the adult taste system and I do not want to disrupt the initial development of taste buds, I will use a tissue-specific conditional knockout mouse and knockout BDNF expression only in the tongues of adult mice. I can also denervate the taste buds in the tissue-specific conditional BDNF knockout mice to see whether nerve fibers will regenerate and reinnervate taste buds that no longer express BDNF. This aim will determine the role(s) of BDNF in adult mouse taste cells, without affecting the initial development of taste buds.

本研究的目的是利用BDNF- lacz转基因“敲入”小鼠来确定BDNF在成年小鼠味觉细胞中的作用。第一个目标是确定成年小鼠中哪些味觉细胞类型表达BDNF。具体来说，BDNF是否存在于新分裂的味觉细胞、未成熟的味觉细胞或表型成熟的味觉细胞中。如果BDNF存在于成熟的味觉细胞中，我感兴趣的是确定BDNF存在于哪些免疫组织化学和形态学类别的味觉细胞中。我将使用双标记免疫组织化学来实现这一目标。为了使细胞诞生，我将给老鼠注射5‘-溴-2’脱氧尿苷它能融入分裂细胞的DNA中。我也有兴趣确定味蕾神经支配的丧失对BDNF的表达有什么影响，以及随后的味蕾再生和神经支配对BDNF的表达有什么影响。为了实现这一目标，我将通过手术粉碎BDNF- lacz小鼠的味觉神经，并让这些小鼠在神经粉碎后连续生活更长的时间，以观察神经退化和再生对味蕾中BDNF表达的影响。对于我的最终目标，我感兴趣的是确定成人味蕾中BDNF表达的丧失对味蕾、味觉神经纤维和味觉神经节细胞有什么影响。由于我对BDNF表达缺失对成年味觉系统的影响很感兴趣，而且我不想破坏味蕾的初始发育，所以我将使用组织特异性条件敲除小鼠，只敲除成年小鼠舌头中的BDNF表达。我还可以切除组织特异性条件BDNF敲除小鼠的味蕾，看看神经纤维是否会再生，并重新支配不再表达BDNF的味蕾。这一目标将确定BDNF在成年小鼠味觉细胞中的作用，而不影响味蕾的初始发育。