<p>Epilepsy is comorbid
with cognitive and psychiatric dysfunctions. This pathophysiology, associated
with hippocampal synaptodendritic structural and functional changes, is
exacerbated by prolonged seizures (status epilepticus; SE). We found a
correlation between hippocampal dendritic loss and microgliosis after SE, along
with hyperactivation of the classical complement pathway (C1q-C3). These
paralleled increased seizure frequency and memory deficits in a rat model of SE
and acquired epilepsy. C1q leads to C3 cleavage into biologically active
fragments C3a and C3b. Evidence suggests that C1q and C3b contribute to
synaptic stripping by microglia in the developing brain and neurodegenerative
disorders. Thus, we hypothesized that SE-induced C3 activation may alter
hippocampal synaptic protein levels thereby promoting memory deficits. </p>
<p>To
test the hypothesis, different groups of wild type (WT) or C3 deficient (C3KO)
mice were injected with pilocarpine (350mg/kg) to induce SE or saline
(controls): WT-C, WT-SE, C3KO-C, and C3KOSE. At two weeks after SE, mice were
subjected to novel object recognition (NOR) to evaluate recognition memory, and
Barnes maze (BM) to measure hippocampal-dependent spatial learning and memory.
Following behavioral testing, mice were sacrificed and hippocampi collected at
either 2 or 5 weeks after SE to measure changes in C3 protein levels and levels
of synaptic proteins including PSD95, Vglut1, and Vgat. As a method of
verifying our findings, we used a second model of pilocarpine-induced SE in
male Sprague Dawley rats. Starting at 7 days after SE, rats were treated with
cobra venom factor (CVF; 100ng/g, i.p.) or vehicle (veh) every third day. On
days 14-15 rats were subjected to open field and NOR to measure anxiety and
recognition memory. On day 16, rats were sacrificed and hippocampi collected
for western blotting.</p>
<p>WT
and C3KO mice were able to reach stage 4.5-6 seizures after pilocarpine
injections. In NOR trial 1, exploration time for both objects was similar in
all groups (<i>p</i> > .05). In trial 2,
WT-C and C3KO-C mice spent more time exploring the novel object than the
familiar one (<i>p</i> < .05) while WT-SE
mice explored both objects equally (<i>p</i>
> .05). Interestingly, C3KO-SE mice spent more time with the novel object
similar to controls (<i>p</i> > .05),
suggesting that the deficit in object recognition memory induced by SE was
attenuated in C3KO mice. Similarly, veh- and CVF-treated control rats spent
more time exploring the novel object during trial 2 (<i>p</i> < .05). The veh-treated SE rats did not show significant
preference for the novel object versus familiar (<i>p</i> > .05), whereas the CVF-treated SE rats explored the novel
object significantly more than the familiar (p < .05). These findings
support that C3 inhibition after SE prevents recognition memory deficits.
Furthermore, there was a reduction in synaptic proteins PSD95 and Vgat in the
SE-veh group compared to the C-veh group. This difference was not observed in
the C-CVF and SE-CVF groups, suggesting that blocking C3 after SE is
neuroprotective against hippocampal synaptic loss.</p>
<p>Taken together, these findings are the first to show an
association between C3 activation and hippocampal and cognitive deficits in two
rodent models of SE and acquired TLE. We found that depletion of C3 is
sufficient to attenuate SE-induced deficits in NOR-evaluated recognition memory
and changes in the levels of an inhibitory synaptic protein. In conclusion, our
data suggest that SE-induced complement C3 activation contributes to
hippocampal synaptic remodeling and impairments in recognition memory, and that
the complement C3 may be a potential therapeutic target for the memory
comorbidities associated with SE. Future studies will determine the effect of
C3 inhibition on spontaneous recurrent seizures, and whether C3-guided and
microglial-dependent phagocytosis is an underlying mechanism for the SE-induced
epileptogenic synaptic remodeling.</p>
| Identifer | oai:union.ndltd.org:purdue.edu/oai:figshare.com:article/8023367 |
| Date | 15 May 2019 |
| Creators | Nicole D Schartz (6620009) |
| Source Sets | Purdue University |
| Detected Language | English |
| Type | Text, Thesis |
| Rights | CC BY 4.0 |
| Relation | https://figshare.com/articles/THE_ROLE_OF_COMPLEMENT_C3_IN_THE_HIPPOCAMPAL_PATHOLOGY_OF_STATUS_EPILEPTICUS/8023367 |
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