Anti-CALML5 monoclonal antibody, clone 3G21

Conjunctivochalasis (CCH) is a common ocular disease, especially in aged people. However, the molecular mechanism of CCH on transcriptional level has been unclear. In this study, we characterized the transcriptional landscape of human conjunctiva and compared the transcriptome between normal persons (n = 10) and CCH patients (n = 11). Illumina RNA sequencing (RNA-seq) was performed to obtain transcriptional data, and these data were analyzed using various bioinformatics methods, including read mapping, the analysis of gene expression, gene ontology (GO) analysis and Kyoto Encyclopedia of Gene and Genomes (KEGG) metabolic pathway analysis. Additionally, expression patterns of 20 dysregupated genes were validated by qRT-PCR. RNA-seq result showed that clean ratios of 21 samples were more than 95% and more than 92% of all clean reads (32-41 million reads) were mapped to human genome sequence. There were 175 up-regulated genes and 582 down-regulated genes identified in CCH compared to normal persons. Among down-regulated genes in CCH, many genes were related with cell cycle and proliferation, such as BUB1, CCNB1, CCNB2 and CENPA, which might disturb cell growth and proliferation. In addition, several down-regulated genes were associated with keratinization and differentiation of epidermal cells, such as SPRR1A, SPRR1B, and CALML5. In over-expressed genes, CALML6 might play important roles on the development of CCH. The results of qRT-PCR confirmed the accuracy and credibility of RNA-Seq analysis. This study provided a lot of valuable information about pathogenic mechanism of CCH, which could be used to better study CCH in the future.

The structural component of wool and hair fibers, such as keratin-associated proteins (KAPs), has been well described, but the genetic determinants of fiber diameter are largely unknown. Here, we have used an iTRAQ-based proteomic approach to investigate differences in protein abundance among 18 samples from sheep and goats across a diverse range of fibers. We identified proteins with different abundance and are associated with variation in fiber features. Proteins with different abundance are mainly keratin or keratin-associated proteins (KRTAP11-1, KRT6A, KRT38), or are related to hair growth (DSC2, DSG3, EEF2, CALML5, TCHH, SELENBP1) and fatty acid synthesis (FABP4, FABP5). RNA-seq further confirmed the functional importance of the DSC2 gene in the determination of woolly phenotype in goat fibers. This comprehensive analysis of fibers from major fiber producing animals is the first to provide a list of candidate proteins that are involved in fiber formation. This list will be valuable asset for future studies into the molecular mechanisms that underlie fiber diversity. Biological significance: Proteins are the basis for animal-derived hair fibers, yet proteins conferring fiber structure and characteristics in sheep and goats are largely elusive. By examining 27 fibers samples representing 9 fiber types from sheep and goats through the iTRAQ approach, we show a list of differentially abundant proteins that are important to hair structural component, or genes related to hair growth and fatty acid synthesis. RNA-seq further validated the DSC2 gene is key to the woolly/straight hair phenotype in goats.