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Reconstruction
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Tomographic reconstruction of presynaptic terminals in hippocampal area CA3 stratum lucidum from an adult transgenic mouse over-expressing human wild-type alpha-synuclein showing synaptic-vesicle filled terminals associated with convoluted membranous structures.
Full resolution image description
Tomographic reconstruction of presynaptic terminals in hippocampal area CA3 stratum lucidum from an adult transgenic mouse over-expressing human wild-type alpha-synuclein showing synaptic-vesicle filled terminals associated with convoluted membranous structures.
File format
MRC

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Attribution Only: This image is licensed under a Creative Commons Attribution License. View License Deed | View Legal Code

CCDB:73215*
Project: P20094
Project name
Molecular and cellular investigations of alpha-synuclein in Parkinson pathogenesis
Description
The goal of this project is to better understand the role of alpha-synuclein in the progression of PD by applying labeling systems developed at NCMIR to track, at multiple resolution scales, the distribution and trafficking of these proteins associated with Parkinson's disease.
Leader(s)
Daniela Boassa
Collaborator(s)
Monica L. Berlanga
Mary Ann Yang
Masako Terada
Junru Hu
Eric A. Bushong
Minju Hwang
Eliezer Masliah
Julia M. George
and Mark H. Ellisman
Start date
unspecified
End date
unspecified
 
Experiment
Experiment ID
9643
Title
Expression of human-alpha-synuclein in transgenic mice
Purpose
Investigate distribution of over-expressed alpha-synuclein in transgenice mice
Experimenter(s)
Daniela Boassa
Microscopy product
Microscopy product ID
73215
Instrument
JEOL JEM4000EX-1
Microscopy type
TEM
Product type
DOUBLE TILT
Image basename
mouse81_500nm_stratLucid_tomo2
Spatial Axis Image Size Pixel Size
X 3488px 1.04 nm/pixels
Y 3488px 1.04 nm/pixels
Subject
Species
Mouse
Scientific name
Mus
Strain
Line D, PDGF-human alpha-synuclein
Group by
Transgene
Treatment
Transgenic (Tg) mice (n = 3) overexpressing human wild-type ¿-synuclein (PDGF¿human ¿-synuclein Tg mice, line D, from Masliah et al., 2000) and their littermate wild-type controls (n = 3), 10- to 17- month-old males, were used in this study.
Age
17 months
Age class
Adult
Tissue section
Anatomical location
Brain, hippocampus CA3
Thickness
0.5 µm
Specimen description
Organ
Brain
System
central nervous system
Imaging parameters
Type
Electron microscopy product
Recording medium
4K CCD
Magification
15000
Accelerating voltage
400 KV
Specimen preparation
Protocol used
Subjects.Transgenic (Tg) mice (n = 3) overexpressing human wild-type -synuclein (PDGFhuman -synuclein Tg mice, line D, from Masliah et al., 2000) and their littermate wild-type controls (n = 3), 10- to 17- month-old males, were used in this study. The mice were housed in an accredited animal care facility (Association for Accreditation of Laboratory Animal Care International) on a 12 h light/dark cycle with food and water provided ad libitum. Animal care was in accordance with the Guide for Care and Use of Laboratory Animals (NIH publication 865-23, Bethesda, MD) and approved by the Institutional Animal Care and Use Committee.Tissue preparation.Animals were deeply anesthetized with an intraperitoneal injection of pentobarbital (10 mg/g body weight). Next, mice were perfused transcardially with oxygenated Ringer's solution at 37C (0.79% NaCl, 0.038% KCl, 0.20% MgCl26H2O, 0.018% NaHPO4, 0.125% NaCHO3, 0.03% CaCl22H2O, 0.20% dextrose, and 0.02% xylocaine) for ~30 s, followed by 0.1 m PBS, pH 7.4, containing 4% paraformaldehyde and 0.1% glutaraldehyde (37C) for 10 min. The mouse brain was removed and stored in same fixative for 2 h. The brains were divided into hemispheres, and coronal sections were collected using a Vibratome (VT1000E, Leica Microsystems). The left hemisphere was sectioned into 100-m-thick sections for conventional EM, 250-500 nm sections for electron tomography, and processing for serial block face scanning EM (SBEM). The right hemisphere was sectioned into 70-m-thick sections, and serial sections were placed in cryoprotectant for immunohistochemistry.
Imaging product type
Type
Double tilt
X min range
60 degrees
X max range
-60 degrees
X tilt increment
2 degrees
Y min range
60 degrees
Y max range
-60 degrees
Y tilt increment
2 degrees
Imaging product type
Type
Single tilt
Description
singlet_desc
Min range
0 degrees
Max range
60 degrees
Tilt increment
1.5 degrees