Taekwan Lee1, Taeyi You2,3, Geun Ho Im3, Seong-Gi Kim2,3,4, Sungkwon Chung5, and Jung Hee Lee2,3,6
1Korea Brain Research Institute, Daegu, Korea, Republic of, 2Biomedical Engineering, Sungkyungkwan University, Suwon, Korea, Republic of, 3Center for Neuroscience Imaging Research, Institute for Basic Science, Suwon, Korea, Republic of, 4Intelligent Precision Healthcare Convergence, Sungkyungkwan University, Suwon, Korea, Republic of, 5Physiology, Sungkyunkwan University, Suwon, Korea, Republic of, 6Radiology, Sungkyungkwan University, Suwon, Korea, Republic of
Synopsis
It is well known environmental factors can
affect brain plasticity in humans, yet finding strong correlative factors is
difficult due to the long development and complexity of human research. Mouse
enrichment studies allows for better controlled research and by combining it
with fMRI, makes mapping brain-wide plasticity changes possible. Here, we
treated mice into three groups of enrichment, standard caging, and isolated
caging to see how their brain responds to multiple-sensory stimulations. We
found the enrichment group responded stronger in multimodal midbrain and thalamic areas. The isolated group responded less suggesting
mouse fMRI is viable in detecting plasticity changes.
Introduction
Functional MRI has become an increasingly popular tool to investigate
brain function for translational neuroscience. Studies investigating the
functional changes due to disease or functional role of certain nuclei within
the brain have been well explored with fMRI in humans and rodents. Various
brain imaging studies have shown functional changes in humans, particularly
children, who grew up in different environmental conditions. This suggests fMRI
is capable in detecting subtle brain plasticity changes caused by environmental
factors, yet whether these changes are detectable with rodent fMRI have yet to
be shown. Rodent studies utilizing an “enriched” environment have shown
cellular, molecular, and behavioral improvements in both wild-type and diseased
models1. An “enriched” environment offers novel stimuli that can promote
somatosensory, motor, visual, and cognitive plasticity compared to the standard
caging mice are kept. As environmental modulation is a potential therapeutic
treatment option for many brain disorders, obtaining in-vivo brain-wide
functional changes with mice fMRI can help supplement its progress and
understanding on brain plasticity. To investigate if such changes are
observable, we utilized a multi-sensory stimulation approach where we attempt
to see whisker-pad, forepaw, visual, and olfactory response in mice that have
been exposed to an enriched environment, standard, or isolated cagings. Methods
Thirty-two C57BL/6 mice were used to investigate the effects of the
housing environment via fMRI. Twelve
mice (in groups of 4) were housed in an “enriched” environment (EE) in which
mice had free access to a 30cm long tunnel, an igloo nest and two running
wheels in an enlarged 50x50cm space. Twelve mice (in groups of 4) were kept in
standard laboratory cages (SC) of 30x22cm. Eight mice were isolated into
individual mice cages (IC) of 10x30cm. All mice were put into their respective
environments post-weaning at 4-weeks of age and kept there for 6 weeks before
fMRI experimentation. Multi-sensory stimulation fMRI experiments were conducted
at 15.2T/11cm Bruker Biospec System under continuous i.v. ketamine/xylazine
anesthesia described previously2. Left forepaw (FP) was electrically
stimulated subcutaneously with 0.6mA at 4Hz, and right whisker-pad (WP) was
electrically stimulated with 0.4mA at 4Hz. Binocular visual stimulation (VIS)
was conducted with white LED lights at 4Hz. Olfactory stimulation (OLF) was
conducted with 5% amyl acetate mixed in mineral oil delivered through the O2/air
gas mixture. Mice were stimulated in block design of 40s baseline, 20s
stimulation, 60s interstimulus interval, 20s stimulation, and 60s recovery.
Stimulation types were randomly done until BOLD response in the primary
cortical area of each respective modality was acquired. Images were
preprocessed and analyzed via GLM.
BOLD amplitude from area-under-curve (AUC) was calculated by averaging the area
of the two response peaks. Results
All stimulation modalities resulted in expected activation in the
respective sensory axis2,3. OLF stimulation resulted in activation in the
main olfactory bulb (MOB) and piriform cortex (PIR) along with downstream
regions such as prefrontal cortex, medial dorsal thalamus (MD), and hippocampal
formation (Figure 1). Olfactory stimulation did not have any significant
differences among EE, SC, and IC groups. With WP stimulation, significantly
higher response in the midbrain reticular nucleus (MRN) and substantia nigra
pars reticulata (SNr) in the midbrain, and ventral posterior (VP), posterior
(POm), subparafasicular area (SPA), and ventral anterolateral (VAL) in the
thalamus were observed in EE compared to SC and IC (Figure 2). FP stimulation
resulted in significantly higher response in the ventral retrosplenial cortex
(RSCv) of the EE group (Figure 3). VIS stimulation resulted in significantly
lower activation in the IC group in lateral dorsal (LD) and lateral posterior
(LP) thalamus, posterior parietal cortex (PPCa), and primary and secondary
visual cortices compared to EE and SC (Figure 4). Discussion
EE group presented with higher BOLD activity in midbrain (MRN, SNr) and
thalamic (VP, POm, VAL, SPA) areas from WP stimulation compared to SC and IC
groups. MRN contains high connectivity to the thalamus and cortex and modulates
ascending sensory signals. SNr is involved in motor execution. The enhanced
thalamic responses were found to be involved in cortico-thalamo-cortical
modulation of the sensorimotor pathway4. Similarly, FP stimulation resulted
in higher activity in the RSCv in the EE group which is known to be a hub for
integrating sensorimotor signals5. The enhanced activity in EE suggests
functional plasticity changes within the sensorimotor pathway due to the
increased movement and exploration. Contrarily, IC group presented with lower
BOLD activity from VIS stimulation within the visual cortex (V1/V2) and
thalamus (LD/LP), along with reduced activity in the PPCa which is involved in
coordinating action from multimodal sensory inputs. This may be due to the lack
of visual and sensory input for the IC group
which may have resulted in reduced function in these cortical areas. The lack
of difference from OLF stimulation may be due to not providing novel olfactory
stimuli for our EE group.Conclusion
We have shown the feasibility in detecting functional plasticity
changes brought upon by environmental conditions in which mice are housed with
fMRI. We see a possible gain-of-function in midbrain and thalamic nuclei in the
EE group related to sensorimotor pathways. A loss-of-function was observed in
the IC group in response to VIS stimulation. These nuclei may be potential
targets for future studies regarding enrichment plasticity.Acknowledgements
This research was supported by the National Research Foundation of Korea (NRF) funded by the Korean government (NRF-2020R1A2C2012416).References
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