Name: Confocal imaging of excitatory synapses in Basolateral amygdala of C57BL6/J mouse injected with PBS, synuclein monomer, or synuclein fibrils
Published: 2024

Confocal imaging of excitatory synapses in Basolateral amygdala of C57BL6/J mouse injected with PBS, synuclein monomer, or synuclein fibrils

[ DOI: https://doi.org/10.35077/g.1168 ]

Dataset Citation:

Volpicelli-Daley, Laura, Gcwensa, Nolwazi. (2024). Confocal imaging of excitatory synapses in Basolateral amygdala of C57BL6/J mouse injected with PBS, synuclein monomer, or synuclein fibrils. [ Collection / Dataset ]. Brain Image Library. https://doi.org/10.35077/g.1168

TechnicalInfo:

File naming conventions used: All components are separated by an underscore. 1. First 3 characters represent what the mouse was injected with - PBS(phosphate buffered saline), PFF (alpha-synuclein pre-formed fibrils), or MON(monomeric alpha-synuclein) 2. The first number represents the mouse ID 3. The age in weeks post injection that the mouse was perfused 4. The person performing the injection 5. The location scanned - basolateral amygdala (BLA) 6. Primary antibody used (vglut2, psyn, homer) 7. Secondary antibody used to conjugate to (488, 555, 647) 8. Primary antibody used(vglut2, psyn, homer) 9. Secondary antibody used to conjugate to (488, 555, 647) 10. Primary antibody used(vglut2, psyn, homer) 11. Secondary antibody used to conjugate to (488, 555, 647) 12. objective (60X) "z2.06" is standard within all file names 13. The next 3 numbers refer to the frame number (~10 frames captured per mouse) 14. deconvolved 40 iterations, typericharson-lucy means the deconvolution method 15. ZXX is the slice in the Z stack (typically around 20 slices total) 16. The last component is the channel ( Cy5 = alexa 647, FITC = alexa 488, TRITC = alexa 555, Dapi = Hoechst staining)

Abstract:

This dataset consists of images from a study determining if fibril-induced alpha-synuclein inclusions cause synapse morphology changes in the basolateral amygdala. Mice received bilateral injections of alpha-synuclein pre-formed fibrils into the striatum. Phosphate buffered saline or monomeric alpha-synuclein were injected as controls because neither of these produce alpha-synuclein aggregates. Six or twelve weeks later, mice were perfused and immunofluorescence was performed for pSer129-alpha-synuclein to visualize inclusions and either vGLUT1/Homer one for corticostriatal synapses or vGLUT2/Homer for thalamocortical synapses. Confocal Z-stacks were acquired, deconvolution was performed, and Imaris was used to render surfaces. A threshold was also applied to filter out any vGLUT1, vGLUT2, or Homer puncta that were not paired in a synaptic foci. The data showed no changes in excitatory synapse density at any time point after PFF injections into the striatum. Both vGLUT1/Homer and vGLUT2/Homer synaptic loci localized close to pSyn aggregates show increased volumes.

Methods:

The BLA was identified using coronal sections from Bregma levels -1.22mm and -1.94. Confocal images were collected with laser power, gain, offset and pinhole diameter kept constant. Z-stack sections were 0.125 micrometer and about 20 were collected per frame. The Z stacks were deconvolved using the richardson-Lucy algorith. Imaris was used to generate surfaces.

Funding:

National Institute of Neurological Disorders and Stroke 1-R56-NS117465-01 Alpha-synuclein aggregate induced synapse loss is a pathological event contributing to Lewy body dementias;
American Parkinson's Disease Association 977962 mGluR4 positive allosteric modulator to inhibit amygdala defects caused by alpha-synucleinopathy;
Aligning Science Across Parkinson's Disease 020616 Understanding and Manipulating Cellular and Circuit-Level Vulnerability to Neurodegeneration in Parkinson’s disease

Contributors:

Volpicelli-Daley, Laura (ProjectLeader) [ORCID: https://orcid.org/0000-0001-8934-4018 ] University of Alabama Birmingham
Gcwensa, Nolwazi (DataCollector) [ORCID: https://orcid.org/0000-0001-9708-6119 ] University of Alabama Birmingham

Related Identifiers: