Sleep Deprivation In Hippocampal Proteomics

Objectives:Sleep is very important in human life.Sleep deprivation(SD) is due to the needs of modern life or work, the environment or other inevitable reasons thus can not guarantee normal sleep.The impact of SD on cognitive function in humans can not be ignored,including decreased awakeness,alertness and attention,reduced learning and memory,declined executive function,increased negative emotions,etc.These series cognitive function disorders and some important changes in the functional areas of the brain or its function related proteins changes caused by the lack of sleep are closely related.After SD in the hippocampus can be observed many changes(replacement,apoptosis, dendrites and synapses reconstruction of neurons),and in the prefrontal cortex and amygdala in rodents can also be observed some other adaptive change. Hippocampal neurons can be observed obvious damage after SD,and alteration of neuronal morphology directly affects their normal physiological function. SD can cause neuronal apoptosis in rat hippocampal.The hippocampus that is a part of the brain located in the temporal lobe,which forms part of the limbic system,plays important role of memory and spatial orientation.Hippocampal in patients with senile dementia is also the site of an early pathological changes.Studies suggest that SD is closely associated with many neurodegenerative disorders,and a variety of neurodegenerative disorders are found to have gathered some pathogenic proteins.In summary,the research directly proceeded on SD that induced proteome changes of the brain tissue may be more intuitive to reveal changes of the body systems especially in the nervous system which causes changes of the pathophysiological mechanisms.As everyone knows,proteins are the final executor of gene expression, they are also the embodiment of vital complexity and diversity. The technology of proteomics is the sign of which life has entering into post-genome era.It intergrally studies the feature of the proteins,containing the expression of proteins,the decoration of proteins′post-translation,interaction between proteins,and et al,in order to explain the rule between protein functions and life.While,differential proteomics is the method to integrally analyze the difference of protein expression in distinct organism at different condition,which can provide the possibility for the research of molecular mechanism in cell and tracing treat drone.In this paper,we focused on the differential proteomics of normal hippocampus tissue and unusual ones which have suffered with SD for 72 h by manufacturing model of SD in rats.In this way,we expect to identify the expression of different proteins,so after exploring the pathophysiology of SD caused a series of changes of the protein,the specific function for further screening for drug-related proteins and intervene to provide dues.Methods:1 Experimental groups: Adult male Wistar rats were divided into experimental group and control group, the experimental group of rats deprived sleep for 72 h and in the control group of rats to maintain normal sleep-wake cycle.2 After rats model of SD, rats on SD experiments were sacrificed after 72 h, the brains were removed and isolated brain hippocampus; the control group, the same parts of the brain to take appropriate, the organization put in place after the EP tube-80℃spare refrigerator.3 Hippocampal tissue proteins were extracted and digested with trypsin after desalting.4 The prepared samples labeled peptides by TMT, using buffer dissolution, strip hydration, and after TFA addification C18 zip-Zip desalination.5 After the sample was dissolved drained, centrifuged, and dried LC-LTQ-MS/MS analysis and mass spectrometry data retrieval.Results:After successfully produced the model of sleep deprivation in rats,using proteomics and TMT label technique,we found 99 kinds of differencial proteins when comparative 72 h of SD group and normal control group in hippocampal tissue,of which 65 kinds of proteins expression levels are upgrade and 34 kinds downgrade.We find more than 60 kinds of meaningful proteins using Gene Codis analysis,those proteins involve in most biological processing, including protein processing and degradation,synapse regeneration, oxidative stress,cell cycling,metabolism,apoptosis etc.Conclusion:1 Using proteomics and TMT label technique to identify proteins,we find those proteins involve in protein expression,processing and degradation,regeneration and synapse function,oxidative stress,apoptosis difference compared with the control group,the hippocampus after sleep deprivation,etc.Most of these processes are closely related to the occurrence of degenerative diseases of the nervous system especially in the pathogenesis of AD.Lack of sleep may be a potential link tips with brain aging and neurodegenerative disease.To help us further understand the nature of these diseases.2 This experiment show that SD leads to proteins changes in brain hippocampal tissue of rats,these proteins are involved in the occurrence and development of degenerative diseases of the nervous system and further indicates that SD is closed with neurodegenerative diseases.But these diseases is a chronic degenerative process and this experiment as an acute stress factor, whether the proteins will be removed after recovery sleep while the stress change is still not very clear. So the relationship between SD and degenerative diseases of the nervous system is still confirmed by further studies.