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 研究中心，而研究中心不一定是研究者所在的机构。 年龄相关的功能和结构的泪液产生泪腺的变化被认为是由副交感神经支配的逐步减少。目标是确定控制泪腺泪液产生和分泌所必需的靶基因，并阐明随年龄变化的细胞网络和显示神经特异性调节的细胞网络。 使用GeneChip Rat Genome 230 2.0阵列生成了正常衰老、神经消融和泪腺反应过度刺激下年轻和老年大鼠泪腺的全基因组基因表达谱。使用ANOVA（p1.5倍）过滤表达数据。MetaCore系统生物学平台用于识别不同年龄和神经操作下的共同和独特基因。基于先验知识生成表达数据子网络和转录调控子网络，并使用基因本体富集进行解释。通过真实的时间RT-PCR或Western印迹分析确认的显著基因被用作“参考靶标”以评估不同的网络。 随着年龄的增长和神经消融，下调基因（83个转录本）几乎是上调基因（33个转录本）的三倍。下调基因的变化幅度更大。还鉴定了几种可用作评估泪腺功能和推定泪液蛋白的生物标志物的细胞靶点。 由于微阵列数据的局限性，将需要进一步的实验（即ChIP-PCR），以验证涉及泪腺基因对不同神经操作的反应的细胞和转录网络。

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. Age related changes in the function and structure of the tears producing lacrimal gland is postulated to be contributed by progressive reduction in parasympathetic innervation. The goal is to identify target genes that are essential in controlling lacrimal gland tear production and secretion, and to elucidate cellular networks that are altered with age and those that displayed neural-specific regulation. Genome-wide gene expression profile in young and aged rat lacrimal gland under normal aging, neural ablation, and overstimulation of the lacrimal gland response were generated using GeneChip Rat Genome 230 2.0 arrays. Expression data were filtered using ANOVA (p1.5 fold). The MetaCore systems biology platform was used to identify common and unique genes under different ages and neural manipulations. Sub-networks of expression data and sub-networks of transcription regulation were generated based on a priori knowledge and interpreted using gene ontology enrichment. Significant genes confirmed by real time RT-PCR or Western blot analysis were used as "reference targets" to evaluate the different networks. With age and neural ablation, downregulated genes (83 transcripts) were almost three times greater than for upregulated genes (33 transcripts). The magnitude of change was greater for the downregulated genes. Several cellular targets that may serve as biomarkers to assess lacrimal gland function and putative tear proteins were also identified. Due to the limitations of microarray data, further experiments (ie.ChIP-PCR) will be needed in order to validate the cellular and transcription networks that are implicated in lacrimal gland gene response to different neural manipulations.