LACRIMAL GLAND BIOINFORMATICS: A NEURAL CONNECTION OF DRY EYE AND AGING

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 据推测，激活传入感觉通路以影响泪腺反应的能力随着年龄的增长而减弱，并影响基因转录反应。 Fischer大鼠（4和24个月）接受单侧角膜伤口以刺激连续流泪，12小时后，取出泪腺，使用Affyphilus大鼠基因组230 2.0基因芯片进行DNA微阵列分析。 使用RMA方法标准化表达数据并在GeneSpring GX中分析。 应用年龄和创伤因素的双因素方差分析（p0.01），并确定了137个转录本与年龄显著相关，117个转录本与创伤相关，23个转录本显示出与年龄相关的创伤效应。然而，这些转录本的倍数变化幅度明显较小。对于4个月与24个月的创伤，在1.5倍截止值处发现了7种转录物，最显著的是cAMP依赖性蛋白激酶抑制剂β的表达增加。对于4个月与24个月未受伤，发现26个转录本，最显著的是纤连蛋白1和酪蛋白激酶2。 基因本体论分析的过度代表性的分子功能的基因在年龄或创伤的影响，发现转移酶和蛋白转运活性的年龄效应，和创伤的影响电机和连接酶的活性。 整合酶和异构酶活性的相互作用的影响。 12小时后，泪腺中基因反应的幅度作为年龄的函数是非常小的。 在更早的时间点，变化的幅度可能更大，更合适。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. It is hypothesized that the ability to activate the afferent, sensory pathway to affect lacrimal gland response is diminished with age and affect gene transcription response. Fischer rats (4 and 24 months) received unilateral corneal wound to stimulate continuous tearing and after 12 hours, lacrimal glands were removed for DNA microarray analysis using the Affymetrix rat genome 230 2.0 GeneChip. Expression data were normalized using the RMA method and analyzed in GeneSpring GX. The two-way ANOVA (p0.01) for age and wounding factors were applied and identified 137 transcripts that were significant with age, 117 transcripts with wounding, and 23 transcripts that showed wounding effect with age. However, the magnitude of fold change for these transcripts was notably small. Seven transcripts were found at the 1.5 fold cutoff for the 4 vs. 24 month wounded, most notably the increased expression of the cAMP-dependent protein kinase inhibitor beta. For the 4 vs. 24 month unwounded, 26 transcripts were found, most notably fibronectin 1 and casein kinase 2. Gene ontology analysis for over-representation of molecular function for genes in the age or wounding effects found transferase and protein transporter activity in the age effect, and for the wounding effect motor and ligase activity. Integrase and isomerase activity were found for the interaction effect. The magnitude of gene response after 12 hours in the lacrimal gland as a function of age was very minimal. It is probable that the magnitude of change could be greater and more appropriate at an earlier time point.