Resolving the Detailed Structure of Cortical and Thalamic Neurons in the Adult Rat Brain with Refined Biotinylated Dextran Amine Labeling 英文参考文献.docVIP

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Resolving the Detailed Structure of Cortical and Thalamic Neurons in the Adult Rat Brain with Refined Biotinylated Dextran Amine Labeling 英文参考文献.doc

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Resolving the Detailed Structure of Cortical and Thalamic Neurons in the Adult Rat Brain with Refined Biotinylated Dextran Amine Labeling 英文参考文献

ResolvingtheDetailedStructureofCorticaland ThalamicNeuronsintheAdultRatBrainwithRefined BiotinylatedDextranAmineLabeling ChangyingLing1,MichaelL.Hendrickson2,RonaldE.Kalil3* 1DepartmentofSurgery,SchoolofMedicineandPublicHealth,UniversityofWisconsin-Madison,Madison,Wisconsin,UnitedStatesofAmerica,2W.M.KeckLaboratory for Biological Imaging, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America, 3Department of OphthalmologyandVisualSciences,SchoolofMedicineandPublicHealth,UniversityofWisconsin-Madison,Madison,Wisconsin,UnitedStatesofAmerica Abstract Biotinylateddextranamine(BDA)hasbeenusedfrequentlyforbothanterogradeandretrogradepathwaytracinginthe centralnervoussystem.Typically,BDAlabelsaxonsandcellsomasinsufficientdetailtoidentifytheirtopographicallocation accurately.However,BDAlabelingoftenhasprovedtobeinadequatetoresolvethefinestructuraldetailsofaxonarborsor the dendrites of neurons at a distance from the site of BDA injection. To overcome this limitation, we varied several experimental parameters associated with the BDA labeling of neurons in the adult rat brain in order to improve the sensitivityofthemethod.Specifically,wecomparedtheeffectonlabelingsensitivityof:(a)using3,000or10,000MWBDA; (b)injectingdifferentvolumesofBDA;(c)co-injectingBDAwithNMDA;and(d)employingvariouspost-injectionsurvival times. Following the extracellular injection of BDA into thevisual cortex, labeled cells and axons wereobserved in both corticalandthalamicareasofallanimalsstudied.However,thedetailedmorphologyofaxonarborsanddistaldendriteswas evident only under optimal conditions for BDA labeling that take into account the: molecular weight of the BDA used, concentrationandvolumeofBDAinjected,post-injectionsurvivaltime,andtoningoftheresolvedBDAwithgoldandsilver. In these instances, anterogradely labeled axons and retrogradely labeled dendrites were resolved in fine detail, approximatingthatwhichcanbeachievedwithintracellularl