Table 1.
Rodent models of wild type or ALS-linked mutant TDP-43.
| Investigators | Wegorzewska et al., PNAS 106, 2009 | Stallings et al., Neurobiol Dis 40, 2010 | Xu et al., J Neurosci 30, 2010 Xu et al., Mol Neurodegener 6, 2011 | |||
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| Promoter | Mouse prion protein (Prp) | Mouse prion protein (Prp) | Mouse prion protein (Prp) | |||
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| Transgene | Human flag-TDP-43A315T | Human TDP-43WT | Human TDP-43A315T | Human TDP-43WT | Human TDP-43M337V | |
| Expression levels | ~3× the endogenous murine protein level in spinal cord by non-species-specific TDP-43 antibody | Line 21: significantly higher than the endogenous murine protein level, but lower than the A315T line 23 level | Line 23: 4× the endogenous murine TDP level in spinal cord by non-species specific TDP-43 antibody | Homozygous: 2.5× the endogenous murine protein level in brain by non-species-specific TDP-43 antibody | Line 4, homozygous: 2.7× the endogenous murine protein level in brain by non-species-specific TDP-43 antibody | |
| Phenotype | 0–3 months: normal weight and appearance 4.5 months: weight loss, “swimming” gait Survival: 154±19 days |
10 founders died between 12 and 55 days. Line 21: no motor phenotype up to 11 months | 5 founders died between 14 and 73 days, others can reproduce but die at 52–75 days or have no phenotype. Line 23: declining grip strength and stride length leading to death within 150 days | Hemizygous: similar to nontransgenic. Homozygous: 14 days: lower body weight compared to nontransgenic mice 21 days: hindlimb clasp, body tremors, and “swimming” gait Survival: between 1 and 2 months |
Hemizygous: similar to nontransgenic up through 1 year. Homozygous: 21 days: hindlimb clasp, body tremors, and dragging gait. 1 month: lower body weight compared to nontransgenic mice, early lethality |
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| TDP-43 and ubiquitin localization | Diffuse nuclear TDP-43 in most neurons and glia. In cytoplasm of neurons in cortical layer 5 and ventral horn, increased diffuse ubiquitin and punctate ubiquitin aggregates. Loss of nuclear TDP-43 staining in some ubiquitin-aggregate-positive neurons | Diffuse nuclear TDP-43 in neurons and glia. Some ventral horn or brainstem neurons with increased diffuse cytoplasmic ubiquitin | Diffuse nuclear TDP-43 in neurons and glia. Colocalization of punctate ubiquitin and phosphorylated TDP (409/410) aggregates in cytoplasm and nuclei of ventral horn and brainstem neurons | Diffuse TDP-43 staining in nuclei, and in cytoplasm of some neurons. Increased diffuse nuclear and cytoplasmic ubiquitin staining. ~15% of spinal motor neurons with phospho-403/404 TDP-43 nuclear aggregates. Rarer cytoplasmic pTDP-43 aggregates | Diffuse TDP-43 staining in nuclei, and in cytoplasm of some neurons. Increased diffuse nuclear and cytoplasmic ubiquitin staining. Many spinal motor neurons with phospho-403/404 TDP-43 nuclear aggregates. Rarer cytoplasmic pTDP-43 aggregates | |
| TDP-43 fragments [antibody used] | 25 and 35 kDa fragments (detergent-soluble) in 1–2 months and older mice [Proteintech 10782-2-AP, anti-body to TDP-43N-260aa] | Some low molecular weight fragments in spinal cord homogenate and cytosolic fractions [Proteintech 10782-2-AP, antibody to TDP-43N-260aa] | Low molecular weight fragments in spinal cord homogenate and cytosolic fractions [Proteintech 10782-2-AP, antibody to TDP-43N-260aa] | 25 and 35 kDa fragments in brain and spinal cord extract from hemizygous and homozygous mice [Proteintech 12892-1-AP, antibody to TDP-43 260aa-C] | 25 and 35 kDa fragments in brain lysates from nontransgenic, hemizygous, and homozygous mice [Proteintech 12892-1-AP, antibody to TDP-43 260aa-C] | |
| Other pathology | Astrogliosis (by GFAP staining) | In cortical layer 5 | Mild, in spinal cord | In ventral horn and brainstem | In spinal cord | In spinal cord and brainstem |
| Motor neuron loss | Decreased number of neurons in cortical layer V. 20% loss of motor neurons in ventral horn | Not quantified | Not quantified | Not quantified | Not quantified | |
| Other aggregates | Unidentified cytoplasmic aggregates displacing nuclei in some neurons | – | – | ~10% of spinal motor neurons contain cytoplasmic mitochondrial clusters. | Mitochondrial clusters in many spinal motor neurons | |
| Investigators | Wils et al., PNAS 107, 2010 | Shan et al., PNAS 107, 2010 | ||||
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| Promoter | Murine Thy1.2 | Murine Thy1.2 | ||||
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| Transgene | Human TDP-43WT | Human TDP-43WT | ||||
| Expression levels | Line TAR4 (hemizygous): human TDP transcript level 1× that of endogenous mouse TDP transcript level in brain | Line TAR6/6 (homozygous): human TDP transcript level 1.2× that of endogenous mouse TDP transcript level in brain | Line TAR4/4 (homozygous): human TDP transcript level 2× that of endogenous mouse TDP transcript level in brain | W3 Line, males: 4.6× endogenous murine TDP level in spinal cord at 4 weeks by non-species-specific TDP-43 antibody | W3 Line, females: 2.3× endogenous murine TDP level in spinal cord by non-species-specific TDP-43 antibody | |
| Phenotype | 14 months: hindlimb clasp 15 months: impaired rotarod performance |
2 months: abnormal hindlimb clasp. 4 months: impaired rotarod performance Survival: avg. 6.7 months |
14–26 days: abnormal hindlimb clasp, decreased stride, impaired rotarod performance, rapid onset spastic paralysis, and death. Survival: <30 days |
14–18 days: hindlimb clasp, tremor, gait abnormalities 4 weeks: ~20% lower body weight compared to nontransgenic mice |
0–3 months: normal weight and appearance 3 months: fine tremor |
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| Animals characterized | Line TAR 4/4 | W3 males | |
|---|---|---|---|
| TDP-43 and ubiquitin localization | Intranuclear and cytoplasmic ubiquitin aggregates in neurons in cortical layer V and spinal cord. Diffuse nuclear TDP-43 in neurons, with some clearing in neurons with ubiquitin aggregates. Intranuclear ubiquitin aggregates costain for human TDP-43 and phospho-409/410 TDP-43. Some cytoplasmic ubiquitin aggregates costain for phospho-409/410 TDP-43. | Diffuse nuclear TDP-43 staining in spinal cord neurons. Intranuclear aggregates of human TDP-43 colocalized with FUS/TLS and SC35. Diffuse cytoplasmic ubiquitin staining. | |
| TDP-43 fragments [antibody used] | 35 kDa C-terminal fragments in cytoplasmic and nuclear fractions of brain lysates. 25 kDa fragment in nuclear fraction. [Rabbit anti-C-terminal TDP-43 and Proteintech rabbit anti-TDP-43] | No 25 or 35 kDa fragments observed | |
| Other pathology | Astrogliosis (by GFAP staining) | In cortical layer V and spinal cord | In lumbar spinal cord |
| Motor neuron loss | ~30% cortical layer V neuron loss, ~25% neuron loss in lumbosacral region of spinal cord | Not quantified | |
| Other aggregates | – | Mitochondrial cytoplasmic aggregates displacing nuclei in some motor neurons |
| Investigators | Tsai et al., J Exp Med 207, 2010 | Igaz et al., J Clin Invest 121, 2011 | Swarup et al., Brain 134, 2011 | ||
|---|---|---|---|---|---|
| Promoter | Ca2+/calmodulin-dependent kinase II (CaMKII) | CaMKIIa-tTA×tetOff | CaMKIIa-tTA×tetOff | 18-kb full length gene fragment from human bacterial artificial chromosome | |
| Transgene | Mouse TDP-43 cDNA | TetOff human TDP-43WT induced at 28 days | TetOff human TDP-43ΔNLS induced at 28 days | Human TDP-43WT Human TDP-43A315T Human TDP-43G348C |
|
| Expression levels | Homozygous: 2× endogenous murine TDP level in hippocampus and cortex by non-species-specific TDP-43 antibody | Line 12: 0.8× endogenous murine TDP level in cortex by non-species-specific TDP-43 antibody | Line 4: 7.9× endogenous murine TDP level in cortex by non-species-specific TDP-43 antibody | All lines: human TDP transcript level ~3× that of mouse TDP in spinal cord at 3 months | |
| Phenotype | 2 months: impaired performance in Morris water maze and fear conditioning test 6 months: abnormal hind limb clasp, impaired rotarod test Mean survival: 495 days |
1–3 month post-induction: limb clasping | 1 week post-induction: limb clasping | All lines: deficits in passive avoidance test, Barnes maze test, and rotarod at 7–10 months | |
| TDP-43 and ubiquitin localization | Cytoplasmic accumulation, ubiquitin colocalization, and nuclear clearing of TDP-43 in 15–20% of cortical neurons. Diffuse nuclear TDP-43 in other neurons | Mainly nuclear staining in cortical neurons in a mosaic pattern. <0.1% of cortical neurons have phospho-409/410 TDP aggregates. Neurons expressing human TDP have decreased mouse TDP-43 expression. | Mainly cytoplasmic with some nuclear staining in cortical neurons. <1% cortical neurons have cytoplasmic phospho-409/410 TDP-43 aggregates colocalizing with ubiquitin. Neurons expressing human TDP have decreased mouse TDP-43 expression. | Mutant lines, 10 months: diffuse and some punctate cytoplasmic staining of human TDP-43 with ubiquitin in mutant spinal cord, cortex, and hippocampus. Not found in WT mice or in mutant mice at 3 months. | |
| TDP-43 fragments [antibody used] | In urea-soluble brain extract: increased high molecular weight, 25 kDa, and 35 kDa fragments in 6 month vs. 2 month mice [Protein-tech 10782-2-AP, antibody to TDP-43N-260aa] | No C-terminal fragments detected | No C-terminal fragments detected | Mutant lines: 35 kDa and 25 kDa fragments in spinal cord increased at 10 months vs. 3 months. [not stated] | |
| Other pathology | Astrogliosis (by GFAP staining) | In hippocampus and cortex | In hippocampus and cortex | In hippocampus and cortex | Using a luciferase reporter, GFAP promoter activity elevated by 20 weeks in brain and ~30 weeks in spinal cord |
| Motor neuron loss | 24% neuron loss in cortex | ~20% neuron loss in dentate gyrus 1 month post-induction, and ~75% loss 3 month post-induction | ~50% neuron loss in dentate gyrus 1 month post-induction | No difference in number of axons in L5 ventral root | |
| Investigators | Rat model: Zhou et al., PLoS Genet 6, 2010 | ||
|---|---|---|---|
| Promoter | 22 kb minimal human TDP gene from bacterial artificial chromosome | CAG-tTA×tetOff | |
| Transgene | Human TDP-43WT Human TDP-43M337V |
TetOff Human TDP-43M337V | |
| Expression levels | Not quantified, but WT and M337V mice have comparable levels in spinal cord by human-specific TDP-43 antibody. | Not quantified, but has levels comparable to WT mice in spinal cord by human-specific TDP-43 antibody. | |
| Phenotype | WT: no paralysis within 200 days. M337V: founders became paralyzed before 30 days. | Line 16, no Dox treatment: paralysis and death by P20. Line 16, induced at 4 days before delivery: paralysis by P35. Line 7, induced at 4 days before delivery: weakness by P40, paralysis and death by P55, males more severe than females | |
| Animals characterized | WT line 4 | Line 7 | |
| TDP-43 and ubiquitin localization | Diffuse nuclear and cytoplasmic staining of phospho-409/410-TDP. Diffuse ubiquitin staining | Diffuse nuclear and cytoplasmic staining of phospho-409/410-TDP, rare TDP aggregates in the cortex but not spinal cord. Diffuse ubiquitin staining | |
| TDP-43 fragments | Low molecular weight fragments (35 and 15 kDa) [in-house human-specific TDP-43 antibody] | Low molecular weight fragments (35 and 15 kDa) [in-house human-specific TDP-43 antibody] | |
| Other pathology | Astrogliosis (by GFAP staining) | Not observed | Around spinal motor neurons |
| Motor neuron loss | No loss of motor neurons in L3 ventral horn | ~10–15% loss of motor neurons in L3 ventral horn | |