|
|
|
|
|
CNS Diseases |
|
|
|
The most common central nervous system (CNS) diseases include stroke, traumatic brain injury, drug addiction, Alzheimer¡¯s disease, Parkinson¡¯s disease, anxiety, depression and etc. This group of diseases constitutes one of the biggest social, healthy and financial problems for human being. To fight against these diseases is currently an extensive focus in the fields. By combining experience/expertise from both pharmaceutical industry and academia, we provide valuable services to accelerate the drug discovery for the treatment of these devastating disorders.
|
|
|
Animal models of CNS diseases |
|
|
|
Alzheimer¡¯s disease (AD): |
|
|
|
Genetically modeling in the mouse: Expression of a human mutant gene or a BAC |
|
fragment that is associated with a familial type of AD |
|
|
Pharmacologically/neurosurgically modeling in the rat: Ventricular infusion of β- |
|
amyloid with an Osmotic mini pump |
|
|
|
|
| |
Fig. 1. An AD model in the mouse: A. Wild-type mice. B. AD mice without β-amyloid deposition.
C. AD mice with β-amyloid deposition. D and E. Double staining of β-amyloid deposition. F. I.
and L. Wild-type mice. G and H. Neurofibrillary tangle-like structure in AD mice. J and K. tau
pathology in AD mice. M. Neuronal degeneration and N. tau pathology in AD model mice. |
|
|
|
|
Age-dependent neurodegeneration: |
|
|
|
Conditional double knockout PS1/PS2 in the mouse: which shows age-dependent |
|
neuronal loss and many other AD-like pathologies such as tau |
|
hyperphosphorylation, neurofibrillary tangle-like structure, and dementia |
|
|
|
Parkinson¡¯s Disease (PD): |
|
|
|
Genetic model in the mouse: Expression of a mutant gene that is associated with a |
|
familial autosomal-dominant mutation, or knockout of a gene that is associated |
|
with a familial autosomal-recessive mutation for PD |
|
|
Pharmacological/neurosurgical model in the rat: Unilateral intra-striatal injection |
|
of 6-OHDA to lesion nigro-striatal dopamine neurons |
|
|
Pharmacological/neurosurgical model in the monkey: Systemic administration of |
|
MTPT to lesion dopamine neurons (under development) |
| |
|
 |
|
Fig. 2. PD modeling. A.
Neurosurgery for
microinjection of 6-OHDA in
the rat. B. Forelimb deficits
of contralateral forelimb at 6
weeks after surgery C.
Quantitative analysis of
rotation number after an
Apomorphine challenge.
D. Quantitative analysis of
motor function with a rotarod
test. Data are expressed as
mean ¡À SD.
|
|
|
| |
|
| |
|
Depression: |
|
|
|
Restraint stress in the mouse |
|
|
Tail suspension in the mouse |
|
|
|
Anxiety: |
|
|
|
Genetic model in the mouse: Overexpression of the cholecystokinin receptor-2 |
|
(CCKR-2) in the forebrain |
|
|
Various stress paradigms: early-life stress, chronic stress, or acute stress |
|
|
PTSD model in the mouse: combination of genetic manipulation and |
|
environmental stress |
|
|
|
Schizophrenia/bipolar disorder: |
|
|
|
PCP model in the mouse and rat |
| |
|
Sleep-deprivation in the rat |
|
|
|
Drug addiction: |
|
|
|
Behavioral sensitization |
|
|
Drug of abuse-induced conditioned place preference (CPP) |
|
|
Reinstatement of extinguished CPP |
|
|
|
Amnesic model: |
|
|
|
Various compounds such as ketamine-, scopolamine-induced amnesia in the rat |
|
and mouse |
|
|
|
Mental retardation model |
|
|
|
BDNF knockdown in the mouse |
|
|
|
Stroke/ischemia model: |
|
|
|
Middle cerebral artery occlusion (MCAO) in the rat |
|
|
Forebrain ischemia model in the gerbil |
|
|
|
Traumatic brain injury model |
|
|
|
Weight-dropping in the mouse |
|
|
|
Huntington¡¯s disease model |
|
|
|
Genetic model in the mouse: Expression of a mutant Huntington gene |
|
|
Pharmacological/neurosurgical model in the rat: Striatal microinjection of |
|
quinolinic acid or 3-NY in the rat |
|
|
|
Pain model: |
|
|
|
Nociceptive pain produced by peripheral injection of formalin or capsaicin |
|
|
Inflammatory pain induced by carrageenan |
|
|
Neuropathic pain induced by sciatic nerve ligation, spinal nerve ligation, or |
| |
streptozotocin-induced diabetic neuropathy |
|
|
Postoperative pain induced by a plantar incision |
|
|
|
Animal models for studies of the blood-brain barrier (BBB) |
|
|
|
|
Experimental approaches |
| |
|
|
|
Behavioral studies |
|
|
|
|
Learning and memory: Morris water maze (spatial |
|
|
learning and memory, working memory, reference |
|
|
memory, reversal learning, hippocampus-dependent |
|
|
memory); Barnes maze (spatial learning and |
|
|
contextual conditioning (hippocampus-dependent), |
|
|
cued conditioning (hippocampus-independent), |
|
|
long-term memory, short-term memory, amygdala- |
|
|
dependent memory, fear response |
Fig. 3. Morris water maze |
|
| |
|
|
|
Motor function: Rotarod test, open-field test, walking-beam test, treadmill test |
|
(under development). |
| |
|
Anxiety-like behavior: Elevated-plus maze, fear-conditioning test, open-field |
|
behavior, startle response, and social interaction. |
| |
|
Depression-like behavior: Novelty-induced hypophagia, tail suspension test, |
|
forced swim test, open-field behavior, social interaction, and learned helplessness. |
| |
|
Schizophrenia/bipolar-like behavior: Pre-pulse inhibition (PPI; under |
|
development), working memory, locomotion/motor activity, social interaction, |
|
aggressive/mania-like behavior, forced swim test, and nesting behavior. |
| |
|
Drug addictive behavior: Drug-seeking behavior (self-administration; under |
|
development), behavioral sensitization, conditioned place preference (CPP), and |
|
CPP extinction. |
| |
|
Pain-related behavior: hot-plate, tail flick£¨infrared heat/pressure£© |
| |
|
Homecage behavior and environmental enrichment |
|
|
| |
|
Pharmacological/molecular/biochemical assay |
|
|
| |
|
|
Distributional and quantitative analysis of gene expression at the mRNA and |
|
protein levels: real-time RT-PCR, microarray, in situ hybridization, Western blot, |
|
immunostaining, and ELISA. |
| |
|
in vivo and in vitro isotope-labeled ligand binding/incorporation assay: receptor |
|
binding assay, autoradiography |
|
|
Brain regional drug delivery (stereotaxic microinjection) and chronic brain |
|
regional drug delivery (Osmotic mini-pump micro-infusion) |
|
|
Spatial and temporal epigenetic analysis: histone modification, DNA methylation, |
|
and genome-wide DNA methylation profile. |
|
|
| |
|
Microdialysis (under development) |
|
|
|
|
|
Histological/morphological studies |
|
|
|
|
|
Gross brain morphometry |
|
|
|
|
Colorimetric staining: Nissl |
|
| |
staining (neuron), LFB staining |
|
(glial myelin), Golgi staining |
|
(dendritic spine), Schiff staining, |
|
Fig. 3. Immunostaining. A and B. NeuN
staining shows neuronal loss in a knockout
mouse (B), compared to wild-type mouse (A).
C and D. GFAP staining in the same mice. |
HE staining etc. |
|
|
|
|
mmunostaining, confocal |
|
|
microscope-based double/triple |
|
|
immunostaining (collaboration with local institutes) |
|
|
|
|
Adult neurogenesis; |
|
|
|
|
|
|
Dendritic spine/synapse morphology |
|
|
|
|
|
|
Subcellular fraction |
|
|
|
|
 |
|
|
| |
|
Fig. 4. Confocal analysis of double immunostaining.
A-F. Double immunostaining of NeuN (red) and
DCX (blue) shows cell proliferation. G-I. Double immunostaining of NeuN (red) and BrdU (green)
shows adult neurogenesis in the dentate gyrus. |
Fig. 5. Synaptogenesis/spinogenesis. A-C.
Neurabin-based immunostaining of cerebellar Purkinje¡¯s cell in the mouse. D-G. Golgi impregnation staining of the hippocampus in
the mouse. |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
---------------------- |
|
|
|
---------------------- |
|
|
|
|
|
|
|
|
|
|
