An increasing appreciation of the importance of sleep and the impact of sleep disorders on health has underlined the need for a basic understanding of sleep and sleep regulation. Our proposal uses the model organism Drosophila melanogaster to identify genes that when mutated lead to abnormal sleep. This proposal has three specific aims: 1) To screen lines of transgenic Drosophila created through piggyBac-element mediated mutagenesis for genes which, when mutated, cause an abnormal sleep phenotype;2) To confirm the assignment of the phenotype to a particular piggyBac insertion by examining the phenotype of fly lines in which the transposable element has been precisely excised;3) To perform additional in-depth phenotypic analysis of mutants confirmed in Specific Aim 2. Present techniques for behavioral analysis of Drosophila rely on infrared beam break technology, which has a high error rate in sleep identification in comparison to the video-based screening analysis we have developed. In particular, single infrared beam break technology significantly over estimates sleep bout length and hence the measure of sleep consolidation, making identification of sleep bout length mutants difficult. We propose using video-based methods to analyze the sleep phenotypes of 1,042 lines that were created using the piggyBac transposable element. These lines are unique and represent insertions into genes that are not found previously generated P-element lines. The assignment of the piggyBac insertion as the source of the phenotype of putative sleep mutants will be confirmed and we will molecularly confirm the gene(s) disrupted by the transposable element. Upon confirmation sleep mutants will be subjected to additional behavioral analyses. These analyses include feeding behavior, testing the mutant response to sleep deprivation and assessing of mutant circadian function. We see two areas of research for our future goals. If this proof of concept study is successful, we will screen the additional transposable element insertions for mutant sleep phenotypes. Concurrently, we propose to characterize the involvement of the specific genes identified in sleep/wake control by genomic techniques using a combination of molecular biological and protein analysis tools.
This proposal will address the genetic basis for sleep. Understanding the genetics of sleep is crucial to our understanding of the functions sleep fulfills and the development of novel treatments for sleep disorders, such as hypersomnia and insomnia.
| Zimmerman, John E; Chan, May T; Jackson, Nicholas et al. (2012) Genetic background has a major impact on differences in sleep resulting from environmental influences in Drosophila. Sleep 35:545-57 |
| Raizen, David M; Zimmerman, John E (2011) Non-mammalian genetic model systems in sleep research. Sleep Med Clin 6:131-139 |
| Singh, Neetu; Lorbeck, Meridith T; Zervos, Ashley et al. (2010) The histone acetyltransferase Elp3 plays in active role in the control of synaptic bouton expansion and sleep in Drosophila. J Neurochem 115:493-504 |
