| ObjectiveTubulin is a important part of cystoskeleton which has tight relationship with the maintaining of cell morphous,substance transferring in the cell,cell movement and cell prolifemouseion.Usually depolymerize of tubulin is necessary during the process of caryocinesis to form spindle microtubule. Only with the draught of spindle microtubule chromatosome can move to double poles.After caryocinesis,those microbule renew depolymerize into tubulins.β3 tubulin specially exist in nervi cell and nutrient cell.Because it is the only isomerism pattern in early neurogenesis,β3 tubulin commonly used to mark the differentiation of stem cell.It expresses in both mature and immature olfactory sensory neurons. Through the combination of the 217-231 amino acid residue of intermediate piece and 31 amino acid residue of N-apical inβ3 tubulin, paclitaxel can increase the interaction of the three protofilament which compose microbule,stable tubulin conformation, restrain microbule disaggregation and cause apoptosis.Olfactory sensory neuron is the only direct contact with the outside environment, and life its special rebirth with divide ability. Its important meaning lies in the transplantation repair of treatment and instaumouseion and central nervous system system that makes to the sense of smell function to make possible.This research applied immunity fluorescence, deeply shoot to give or get electric shock a mirror and solid metered RT-Q-PCR method monitor tubulin and smell the expression variety of marking egg white(olfactory marker protein, OMP), inquire into paclitaxel in the process of smelling the sensory nerve dollar dividing in to the function of tubulin; Combine to pass to grow the factor(nerve growth factory, NGF) processing with the nerve, the first step inquires into it to the olfactory protection function. MethodsSPF class mouse, with the male:Is female 1:2 proportions match cage and take to look into Yin to bolt female mouse 30.Random being divided into is two sets of, experiment set and matched control are 15 for each.Experiment set the tail intravenous injection paclitaxel 1 mlthat give the E for 0.5 days since everyday, the matched control gives everyday 1 ml tail intravenous injection in physiology brine.Each one is divided into E9.5, E14.5 and mouse for each in E17.5 again.Applied dual immunity fluorescence, deeply shoot to give or get an electric shock a real-time quantitative polymerase chain reaction(RT-Q-PCR), the method monitor P 3 tubulin whites and smell the expression variety of marking egg white.The combining to apply the immunity fluorescence observation nerve growth factor Yun to expect smells smell the expression variety of marking egg white in the mucosa after the medicine to the small mouse.ResultThe immunity fluorescence result suggests that can seeβ3 particularity dyeings when the E9.5 smelt a board just start developing, the extension expect along with the Yun,βthe particularity of 3 tiny tube egg whites dyes to obviously increase.Have never seen on E9.5 the OMP masculine gender express, E14.5 it is thus clear that the OMP masculine gender expresses, expression strength smallβin same period is 3 tiny tube egg whites, the extension expect along with the Yun, the OMP particularity dyes and obviously increases.After paclitaxel handles, tubulin and OMP expression obviously weaker than and normal matched control, but as for extension that expect along with the Yun, tubulin and OMP expression are to increase gradually. Solid metered RT-Q-PCR result fluorescence dye result and immunity fluorescence result homology.After shooting to give or get an electric shock a mirror to show paclitaxel a processing deeply, the small mouse smells structure mess inside the cell, smells cilia decrease or disappearance and becomes an empty bubble kind change.The small mouse smells the mucosa immunity fluorescence dyeing show the paclitaxel processing set as a resultβ3 tubulin and OMP express decrease, while NGF give medicine after, both expression have to build up, with RT-Q-PCR result homology.DiscussionNerve dollar for middle, but the its expression strength is small theβin same period to isβ3 tubulin, hint to expect still at this time the ex-body cell of existence one fixed amount.The extension that expects along with the Yun, the OMP expression obviously increase, hints to smell a sensory nerve dollar along with the gradual growth of embryo and continuously divides maturity.After paclitaxel handles,β3 tubulin and OMP expression obviously weaker than and normal matched control, hint that the solution that the paclitaxel passes to repress tiny tube egg white gathers, it makes tiny control ability and tiny tube organization center not is mutual to link, repress the normal formation of spindle body, thus repressed the stalk syrup of nerve dollar conveyance, obstruct the growth maturity of smelling the sensory nerve dollar.But expect along with the Yun of extension,β3 tubulin and OMP expression are to increase gradually and explain that the paclitaxel can not completely see a nerve dollar to divide and develop to be held up.NGF passes growth of promoting the nervous system, the growth, survival of maintenance nerve dollar, divide, and influence the plasticity that the Tu touches.Under pathologic status, pass the expression of inducing the nerve growth factor gene, repress the death of nerve dollar, strengthen clearance of free radicals, promote the instaumouseion of nerve dollar function, attain the purpose of protecting the nerve dollar.The experiment shows that NGF gives after the medicine the paclitaxel processing setβ3 tubulin and OMP expression have increment and hint that NGF has certain protection function to the sense of smell function obstacle that the paclitaxel causes as a result.Conclusion1,(33 tubulin express in the earlier period of mouse embryonic development,and with the mature of embryo,its expression obviously enhance. 2,Olfactory sensory neuron dosen't express in the earlier period of mouse embryonic development but in the metaphase, and with the mature of embryo,its expression obviously enhance.3,Paclitaxel restrains the olfactory sensory neuron differentiation and its development byβ3 tubulin.4,Nerve growth factor has certain protection on the damage of mouse olfactory sensory neuron which caused by Paclitaxel. |
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