The Effects Of Volatile Anesthetics On The Behavior Of Paramecium Tetraurelia And Its Mechanism Study

Partâ… The behavioral effects of different concentrations of volatileanesthetics,P485,alkanes and calmodulin antagonists on wild typeParamecium tetraureliaObjective:To examine and compare the viability effects of different concentrationsof volatile anesthetics (ether,enflurane,isoflurane,sevoflurane),isoflurane analogue P485,alkanes (n-pentane,n-decane) and calmodulin antagonists (W-7,calmidazolium chloride)on sodium acetate chemokinesis and the velocity of swimming speed of wild typeParamecium tetraurelia to explore the behavioral effects and its possible mechanism.Method T-Maze was used to investigate the effects of different concentrations of themedicines on sodium acetate chemstaxis of wild type Paramecium tetraurelia.Observingthe dynamic exercise of wild type Paramecium tetraurelia under inverted phase contrastmicroscope,the monitor software Image Pro Discovery was used to take a video record,which was analyzed to get the motion velocity of wild type Paramecium tetraurelia byusing Motion Analysis 1.2.Results:Volatile anesthetics (ether,enflurane,isoflurane,sevoflurane) and calmodulin antagonists (W-7,calmidazolium chloride) dose-dependently reduced the index of chemokinesis of wild type Paramecium tetraurelia but not the P485.Both n-pentane and n-decane changed the index of chemokinesis of wild type parameciumtetraurelia.Volatile anesthetics and calmodulin antagonists in our study temporarily anddose-dependently increased the motion velocity of paramecium and then reduced underbasal level.Both 500mM n-pentane and n-decane temporarily increased the velocity ofswimming speed of wild type Paramecium tetraurelia and then reduced.The differencewas that 500mM n-pentane reduced the motion velocity under basal level,but 500mMn-detane reduced it to zero.However,the isoflurane analogue P485 reduced the velocity ofswimming speed of wild type Paramecium tetraurelia to basal level after temporaryincrease.Conclusion:The wild type Paramecium tetraurelia has satisfactory reflects tovolatile anesthetics.Volatile anesthetics can affect the behavior of wild type Parameciumtetraurelia,and we defined this effect as"Paramecium anesthesia".The wild typeParamecium tetraurelia can be used as an biology mode to study the mechanism of volatileanesthetics.The volatile anesthetics may possiblely influence the wild type Parameciumtetraurelia by Ca²⁺/CaM system.Partâ…¡Volatile anesthetics,P485,alkanes and calmodulinantagonists change intracellular calcium concentration in wild typeParamecium tetraureliaObjective:This study examined and compared the effects of volatile anesthetics(ether,enflurane,isoflurane,sevoflurane),isoflurane analogue P485,alkanes(n-pentane,n-decane) and calmodulin antagonists (W-7,calmidazolium chloride) on wild typeParamecium tetraurelia.Methods:[Ca²⁺]_i of wild type Paramecium tetraurelia wasmeasured by using confocal laser scanning microscope.Changes of calcium concentrationsin the cytosolic space,were determined before and after exposing the cells to volatile anesthetics and calmodulin antagonists.Results:0.8 EC₅₀ volatile anesthetics and lμgcalmodulin antagonists did not change [Ca²⁺]_i of wild type Paramecium tetraureliasignificantly.1 EC₅₀ and 1.2 EC₅₀ volatile anesthetics,and the higher concentrationcalmodulin antagonists significantly increased the [Ca²⁺]_i of wild type Parameciumtetraurelia.The peak [Ca²⁺]_i of wild type Paramecium tetraurelia cell was increased whendrug's concentrations increased.The isoflurane analogue P485 dose-dependently increasedthe peak [Ca²⁺]_i.But there was significant difference between the peak [Ca²⁺]_i inducedby P485 and isoflurane in the same concentration.Conclution:These results suggested thatvolatile anesthetics and calmodulin antagonists all dose-dependently increase [Ca²⁺]_i ofwild type Paramecium tetraurelia.The changes of [Ca²⁺]_i maybe the mechanism of theeffect"motion velocity increase"on wild type Paramecium tetraurelia of these drugs.There was intimate relationship between the mechanism of volatile anesthetics and [Ca²⁺]_i.but the [Ca²⁺]_i changes is not the specificity character of volatile anesthetics.Partâ…¢The increasing of the [Ca²⁺]_i in wild type Parameciumtetraurelia induced by IP₃ uncaging and its relationship with it'smotion characterObjective:To identify the relationship between [Ca²⁺]_i and motion character wildtype Paramecium tetraurelia.Methods:Confocal laser scanning microscope and IP₃uncaging was used to observe the [Ca²⁺]_i in wild type Paramecium tetraurelia successively.The wild type Paramecium tetraurelia incubated with caged IP₃/AM and Fluo-3/AM wastemporary exposured to ultraviolet light filtered from mercury lamp to increase the [Ca²⁺]_iof wild type Paramecium tetraurelia.At the same time,the motiom video of wild type Paramecium tetraurelia was recorded.Results:IP₃ uncaging successfully increased the[Ca²⁺]_i of wild type Paramecium tetraurelia.Meanwhile,IP₃ enhanced the movement ofwild type Paramecium tetraurelia.Conclusion:These results suggest that there should beintimate relationship between [Ca²⁺]_i and wild type Paramecium tetraurelia motioncharacter.Partâ…£The effect of volatile anesthetics,P485,Alkanes andcalmodulin antagonists on affinity of Ca²⁺/CaM systemObjective:To study the effects of volatile anesthetics (ether,enflurane,isoflurane,sevoflurane),isoflurane analogue P485,alkanes(n-pentane and n-decane) and calmodulinantagonists (W-7 and calmidazolium chloride) on the affinity of Ca²⁺/CaM.Methods:Calmodulin was labeled with the amine-specific dye PyMPO-SE[1-(3 (succinimidyloxycarbonyl)benzyl)-4-(5-(4-methoxyphenyl)-oxazol-2-yl)pyridiniumbromide].The glass coverslips,derivatized with aldehyde-terminated silanes for covalentattachment of the protein via surface amines,was optically coupled to a dove prism usingimmersion oil.An argon-pumped Ti:sapphire system operating at 80 MHz with～150 fspulse duration and 0.5 W average power was employed as the fundamental source.Thepolarization of both the fundamental and second-harmonic beams was varied using waveplates.The signal was verified as the second-harmonic by determining its quadraticdependence on the fundamental intensity and measuring its characteristic spectral lineshape.Each data point was obtained by averaging the photon count over 1 s.Results:100μMcalcium ion significantly increased the SHG intensity of CaM.However,volatileanesthetics and calmodulin antagonists dose-dependently attenuated the SHG intensity of Ca²⁺/CaM system,and P485 in our study concentration had no effect in SHG intensity ofCa²⁺/CaM system.500 mM n-pentane significantly attenuated the SHG intensity of Ca²⁺/CaM system but not 500 mM n-decane.Conclusion:Volatile anestheticsdose-dependently inhibited the affinity of Ca²⁺/CaM,and their action mode was identicalto calmodulin antagonists.Anesthesiophore n-pentane also could inhibited the affinity ofCa²⁺/CaM,but not the P485 and n-decane.This study confirmed that Ca²⁺/CaM was theaction target of volatile anesthetics.