ABSTRACT
BACKGROUND: Over the past five years, interest in and use of DNA array technology has increased dramatically, and there has been a surge in demand for different types of arrays. Although manufacturers offer a number of pre-made arrays, these are generally of utilitarian design and often cannot accommodate the specific requirements of focused research, such as a particular set of genes from a particular tissue. We found that suppliers did not provide an array to suit our particular interest in testicular toxicology, and therefore elected to design and produce our own. RESULTS: We describe the procedures used by members of the US Environmental Protection Agency MicroArray Consortium (EPAMAC) to produce a mouse testis expression array on both filter and glass-slide formats. The approaches used in the selection and assembly of a pertinent, nonredundant list of testis-expressed genes are detailed. Hybridization of the filter arrays with normal and bromochloroacetic acid-treated mouse testicular RNAs demonstrated that all the selected genes on the array were expressed in mouse testes. CONCLUSION: We have assembled two lists of mouse (950) and human (960) genes expressed in the mouse and/or human adult testis, essentially all of which are available as sequence-verified clones from public sources. Of these, 764 are homologous and will therefore enable close comparison of gene expression between murine models and human clinical testicular samples.
Subject(s)
Gene Expression Profiling , Oligonucleotide Array Sequence Analysis/methods , Testis/metabolism , Adult , Animals , Hot Temperature , Humans , Male , Mice , Mice, Inbred C57BL , Middle Aged , RNA/genetics , RNA/metabolism , Transcription, GeneticABSTRACT
Advanced glycation end-product(s) (AGE) are formed in biological systems when reducing sugars react with amino groups on proteins. Long-lived proteins such as collagen and lens crystallins are known to be susceptible to AGE modification and may play a major role in the development of diabetes and other age-related pathologies. It has been previously suggested that AGE formation might affect the lifespan of experimental animals. Our study is the first to examine the effect of AGE accumulation on the life span of an organism, Drosophila melanogaster. We found that Drosophila melanogaster maintained at 24 degrees C accumulate significant AGE over their lifespan. Young flies (10 days old) had 44% less AGE than senescent flies (75 days old). We were able to reduce AGE accumulation in Drosophila melanogaster by raising the flies on a medium containing a known AGE inhibitor, aminoguanidine HCl. Reduction of AGE in flies failed to increase their mean lifespan, and high concentrations (40 mM) reduced the mean life span, which suggests that aminoguanidine is toxic at levels near those required for inhibition of AGE formation. However, the common fruit fly, Drosophila melanogaster, provides a simple model system to study the age-dependent accumulation of glycated proteins and their inhibition by novel compounds.
