Exploring The Expression Of Drp1 In The Central Nervous System Of Mice Both In The Level Of Protein And MRNA

Mitochondria are double-membrane organelles that plays a large role in cells by constantly providing energy,determining the fate of cells.Neurons are highly polarized cells with distinct axons and dendrites,and they have a high demand for energy to maintain neural circuits.Studies have shown that mitochondria support multiple aspects of neuronal development and functions by providing cellular energy,maintaining the intracellular calcium levels,generating reactive oxygen species and regulating the release of apoptogenic proteins apoptosis-inducing factor.The dynamic characteristics of mitochondria consisted of the number,quality and distribution are the main mechanism of mitochondrial functions,particularly that the abnormal process of mitochondria fission and fusion has been proved to be involved in the occurrence and development of multiple neurodegenerative diseases.The GTPase protein,dynamin-related protein 1(Drp1)is responsible for the mitochondria fission in the nervous system,and which is different with the other GTPase proteins of GTPase proteins family involved in mitochondrial fusion mechanism.However,the expression of Drp1 in the nervous system,the location of Drp1 in the subcellular(organelle)level,and the distribution of Drp1 in the neurons with different structures are still unclear.The limit in systematic research greatly blocks us clearly recognizing of mitochondrial dynamics changes in the physiological state of nervous system.ObjectiveWe researched brain and spinal cord of mice,exploring the expression of Drp1 protein and Drp1 mRNA in the central nervous system..Answer the following questions:1,Whether the expression of Drp1 is widely,and whether exists regional specificity.2,Whether Drp1 locates in neurons or glial cells,if there is a cell specificity.3,If there is organelles orientation specificity.4,If there is a structure specificity.Materials and MethodsMale C57BL/6 mice and glutamic acid decarboxylase 67-green fluorescent protein(GAD67-GFP)transgenic mice were used.In this study,We used the technologies of western blot(WB),immunofluorescence(IF),immunoelectron microscopy(IEM),fluorescence in situ hybridization(FISH)to observe the expression of Drp1 protein andDrp1 m RNA in the central nervous system.ResultsBoth Drp1 protein and Drp1 mRNA are expressed widely in the central nervous system of mice,especially Drp1 mRNA;Drp1 specifically express in neurons;Drp1 is regional specificity,there is a high level expression of Drp1 in LGd,POR,CN,VII,DR;Drp1 has organelles orientation specificity,most of which exist in the cytoplasm,only a little on the mitochondrial outer membrane.ConclusionDrp1 involves in the a variety of physiological functions through regulating mitochondria fission,which is crucial for maintaining the body's normal functions.The mechanism needs further research.Researches showed that Drp1 mostly distributes in the cytoplasm,only 3% approximately on the mitochondria,which mesns that the molecular basis of Drp1 recruitment to motochondria may be an important part of the research in the future.