63479
Daniela Boassa
purpose
Expression of human-alpha-synuclein in transgenic mice
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512r&mpid=63479]]>
0
Serial block face scanning EM
70 nm
63479
M81_cortex_Vol1
3View
Serial Block Face SEM
SERIAL SECTION
Monica L. Berlanga
Mary Ann Yang
Masako Terada
Junru Hu
Eric A. Bushong
Minju Hwang
Eliezer Masliah
Julia M. George
and Mark H. Ellisman
Daniela Boassa
Molecular and cellular investigations of alpha-synuclein in Parkinson pathogenesis
P20094
Boassa D, Berlanga ML, Yang MA, Terada M, Hu J, Bushong EA, Hwang M, Masliah E, George JM, Ellisman MH. Mapping the subcellular distribution of alpha-synuclein in neurons using genetically encoded probes for correlated light and electron microscopy: Implications for Parkinson's disease pathogenesis. J Neurosci. 2013 Feb 6;33(6):2605-15. doi:10.1523/JNEUROSCI.2898-12.2013. PMID:23392688
Using SBEM we observed, in the neocortex of AS transgenic mice, enlarged nerve terminals massively filled with an endomembrane network formed of tubulovesicular structures.
<![CDATA[http://ccdb.ucsd.edu/sand/main?event=rd&type=512v&mpid=63479]]>
Using SBEM we observed, in the neocortex of AS transgenic mice, enlarged nerve terminals massively filled with an endomembrane network formed of tubulovesicular structures.
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MRC
Three-dimensional volume segmentation shows the enlarged terminal (green) forming a synapse (postsynaptic density is in red) with a dendritic spine (gold) compared to a normal terminal (blue) forming a synapse with an adjacent spine on the same dendrite.
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Brain
central nervous system
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.
Transgene
17 months
Adult
male
Mus
Mouse
Line D, PDGF-human alpha-synuclein
Brain, hippocampus CA3
.5 um
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.