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Neurons
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3-D rotation sequence (5 MB QuickTime file) showing
synapses on developing (P7) rat cerebellar Purkinje Cells.
Purkinje Cells were labeled with antibodies against calbindin (green),
and postsynaptic densities were labeled with antibodies against
PSD-95 (red). Note the nascent synapses (red spots) circumscribing
the cell bodies as well as the developing dendrites. See Castejon
et al., 2004.
Movie by Leah Fuller & Mike Dailey.
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Time-lapse
sequence (5 MB QuickTime file) of calcium transients showing epileptiform
activity in a rat hippocampal slice culture. Neurons and
glia in the slice were loaded with a fluorescent, intracellular calcium
indicator dye, Fluo-3. Images were collected with a confocal microscope
every 7 sec. Note the synchronized (epileptiform) neuronal activity,
which appears as horizontal flashing bands. The bright twinkling cells
are astrocytes. The flashing bands (epileptiform activity) appear
first, followed by strong astrocyte twinkling, which subsides before
the epileptiform activity reappears again. SP=stratum pyramidale,
or layer of pyramidal neuron cell bodies.
Movie by Mike Dailey. |
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Microglia
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Time-lapse sequence
(4.8 MB QuickTime file) showing motile and locomotory microglia moving
among dead cells in a rat hippocampal brain slice culture (P6 + 2
DIV). Microglia were labeled with FITC-IB4 (green) and dead
cell nuclei were labeled with To-Pro-3 (red). A confocal microscope
was used to collect images in two fluorescent channels simultaneously.
Total sequence time is 5 hr.
Movie by Mark Petersen. (See Petersen
& Dailey, 2004. Fig. 4) |
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to play
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Higher magnification
view (2.9 MB QuickTime file) from the same experiment showing dynamics
of microglial motility. Note a mitotic microglial cell (lower
left of center) near the beginning of the time sequence. Also,
near the end of the time sequence, a microglial cell sweeps in from
the upper left corner and picks up a dead cell nucleus "on the
fly".
Movie by Mark Petersen. (See Petersen
& Dailey, 2004, Fig. 4)
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Microglia rapidly
engulf cells that die spontaneously in brain slice cultures
(1.6 MB QuickTime file). Several Sytox Orange-labeled
dead cell nuclei (yellow balls) are already apparent at the
start of this movie. When a cell dies in the presence
of a dead cell marker, its nucleus becomes labeled. In
this movie, a dead cell nucleus appears spontaneously in the
bottom right hand corner of the image. Within minutes,
one of the microglial cells extended a long process directly
toward the dead cell nucleus, engulfed it, and dragged it away.
This demonstrates that activated microglia can sense and respond
very rapidly to cell death events in brain tissues. Rapid
clearance may reduce secondary tissue damage due to release
of toxic products from dead cells. Tissue is P6 + 3 DIV.
Movie by Mark Petersen. (See Petersen
& Dailey, 2004, Fig. 6) |
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Time-lapse sequence
of MG movements in anisomycin treated slices. Five IB4-labeled
MG cells, some of which are closely associated with microvessels,
are evident. Note that after ~4 hr of anisomycin treatment,
few MG are locomoting.
Movie by Dana Kurpius. (See Kurpius
et al., 2006, Fig. 5A) |
Click image to play
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Higher magnification,
time-lapse sequence of MG movements in anisomycin treated slices.
Time-lapse images of a selected MG cell (from sequence
above) showing that, despite a lack of locomotion, the cell
is highly motile, showing many lamellipodial protrusions and
arm-like extensions.
Movie by Dana Kurpius. (See Kurpius
et al., 2006, Fig. 5B) |
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Time-lapse sequence
showing mobile MG near injured pyramidal neurons in tissue slices
from b2 integrin hypomorphic (b2hypo)
mice. The field of view contains the pyramidal cell body layer
(SP) and the adjacent stratum radiatum (SR) and stratum oriens (SO)
in an acutely excised neonatal tissue slice from a b2hypo
mouse. Note that mobile MG cells accumulate in the SP (demarcated
by the dashed lines).
Movie by Dana Kurpius. (See Kurpius
et al., 2006, Fig. 8) |
Astrocytes
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Astrocytes in rat hippocampal slice
cultures develop a complex 3-D morphology. This movie (840K AVI
file) shows a three-dimensional reconstruction of a rat astrocyte
expressing LCK-GFP. A confocal microscope was used to image
the cell in a cultured hippocampal slice.
Movie by Adrienne Benediktsson and Ryan Jeffrey. |
Click image to play
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This is a black and white movie
of an astrocyte that has been transfected, and is therefore expressing
the protein GFP-GLT-1. The bright regions are sites of protein
accumulation or clusters that can be seen moving along the astrocyte's
branches. The movie will begin with a low magnification view, then
zoom up to two smaller regions. The total time represented by this
movie is approximately 5 hours.
Movie by Adrienne Benediktsson. |
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