TABLE 8.
Investigating the effect of O-GlcNAcylation in vivo using thiamet G (22 references). Bold highlights indicate the beneficial effect. Those not highlighted indicate either the detrimental effect or that no biological or functional change was reported.
| Amount used and duration | Observations | References |
|---|---|---|
| Rat (6-week-old Sprague Dawley): i.v. 2, 10, or 50 mg/kg, 4 h | ↑ protein O-GlcNAcylation at 10 and 50 mg/kg in total brain homogenate by western blot with CTD110.6, ↓ pS396 tau/Tau-5 (total) | Yuzwa et al. (2008) |
| Rat (6-week-old Sprague Dawley): i.v. 50 mg/kg, 1–16 h | ↑ protein O-GlcNAcylation in total brain homogenate starting at 1 h and peak at 10 h, by western blot with CTD110.6; ↓ pS396 tau/Tau-5 starting at 1 h and peak at 4 h and restore to normal at 16 h | |
| Rat (21-week-old Long–Evans): drinking water, 200 mg/kg/d, 1 day |
↑ protein O-GlcNAcylation in total brain homogenate by western blot with CTD110.6, ↓ pS396, pT231, and pS422/Tau-5, in total brain homogenate by western blot
↓ pS396 immunostaining in CA1 and cortex; ↓ pS231 immunostaining in the cortex |
|
| Rat (6–8-week-old Sprague Dawley): i.p. 10 mg/kg | ↑ protein O-GlcNAcylation in the hippocampal formation, CA1 pyramidal cells, inhibitory interneurons, and astrocytes 2, 8, or 24 h after injection, by CTD110.6 immunohistochemistry. No change of open-field behavior at 2 and 4 h after injection; no change of contextual fear conditioning 2 h after injection For novel object and placement recognition tests, exploration time at 2 h after injection was unchanged, while the performance in testing was significantly decreased in response to thiamet G |
Taylor et al. (2014) |
| Hemizygous female JNPL3 at 14 weeks of age: drinking water, 500 mg/kg/d, 1 week | ↑ protein O-GlcNAcylation (using CTD110.6 antibody) in the brain; ↑ O-GlcNAcylation on Ser400 of tau/total tau (using an antibody generated against O-GlcNAc-Ser400/HT7 antibody) by western blot analyses | Yuzwa et al. (2012) |
| Hemizygous female JNPL3 mice at 9–12 weeks of age: drinking water, 500 mg/kg/d, ∼30 weeks |
Alleviated body weight loss (administering 500 mg/kg/d thiamet G through drinking water for 22 weeks in wildtype mice did not change body weight, food or water consumption, organ weights, and motor neuron counts).
With the differences in body weight, rotarod and cage hang tests were inconclusive |
|
| Hemizygous female JNPL3 mice at 9–12 weeks of age: drinking water, 500 mg/kg/d, 36 weeks | Alleviated motor neuron loss. Immunohistochemistry studies demonstrated ↑ O-GlcNAcylation, ↓ pTau using AT8 antibody which detects S202 and T205 and S422 phosphorylation, ↓ S422 and Y29 sites of tau phosphorylation using pS422 and nY29 antibodies, and ↓ PHF-1 immunoreactivity | |
| RFP-GFP-LC3 mice at 4–8 weeks for 2 weeks: 500 mg/kg/d thiamet G through drinking water | ↑ red puncta, unchanged yellow puncta, and ↓ p62 autophagic substrate were found in broad brain regions, indicating increased autophagic flux | Zhu et al. (2018) |
| JNPL3 hemizygous female mice at 9–12 weeks of age for 36 weeks: 500 mg/kg/d thiamet G through drinking water | ↑ LC3 puncta, ↑ LC3II, ↓ P62, and unchanged p-MTOR/p-S6K and p-4EBP1 on western blots | |
| 3×Tg-AD mice at 36–40 weeks for 2 weeks: 500 mg/kg/d thiamet G through drinking water | ↓ LC3 puncta and p62 as measured by immunohistochemistry, LC3II was unchanged, and ↓ P62 by western blots | |
| Tau441 largest isoform wildtype human tau (175 µg/mouse, correlating to 700 µM in the brain) i.c.v. at 6 months |
↑ protein O-GlcNAcylation in brain homogenates at 4.5–24 h after injection as demonstrated by western blot using RL2
↓ tau phosphorylation at T181, T212, S214, S262/S356 S404, and S409, ↑ Tau phosphorylation at S199, S202, S396, and S422, and unchanged at T217 by western blot using site-specific antibodies ↓ pS473-AKT, total GSKβ, and pS9-GSK3β were also evident by western blot |
Yu et al. (2012b) |
| Female Thy-1-tau P301L, 4–8 h thiamet G up to 500 mg/kg | ↑ protein O-GlcNAcylation in brain homogenates by western blot using CTD110.6 No change of total or p-tau (Tau5, pS396, pS404, AD2, AT180, or 22E8) |
Borghgraef et al. (2013) |
| Female Thy-1-tau P301L, 3 days drinking water at 2.5 mg/ml | Ameliorated upper-airway breathing defects of these mice | |
| Female Thy-1-tau P301L, 2.5 months drinking water at 2.5 mg/ml | Improved clasping score, body weight, and morbidity | |
| Female double-transgenic APPSwe-Tau mice at 10–23 weeks of age drinking water at 200 or 500 mg/kg/d based on ∼4 ml/d/animal for 34 weeks |
After 20 weeks, the 500 mg/kg/d group exhibited better performance in a Morris water maze probe trial. After 34 weeks, ↑ global protein O-GlcNAcylation in brain homogenates by western blot using either RL2 or CTD110.6, in both 200 and 500 mg/kg/d groups, ↑ immunohistochemical signals in the 500 mg/kg/d group in the hippocampus, cerebellum, pons, and amygdala using RL2 and CTD110.6; total tau or p-tau were unchanged (92e, pS199/pS202 pT205, pT212, pT214, pT217 pT231, pS262/pS356 (12E8), and pS396/pS404 (PHF-1) for western blot; pS396, pS396/pS404 (PHF-1), pT231 (AT180), and pS262/pS356 (12E8) for immunostaining)
↓ Aβ42 in the 500 mg/kg/d group using ELISA assay ↓ plaques in in the cortical and hippocampal regions in the 500 mg/kg/d group and decreased plaque in the cortex in the 200 mg/kg/d group using immunohistochemistry with the 6E10 antibody |
Yuzwa et al. (2014) |
| rTg(tauP301L)4510 mice with acute (1day), subchronic (14days), and chronic (4 months) thiamet G in water administered p.o. at 500 mg/kg/d beginning at 2 months of age |
↑ protein O-GlcNAcylation in brain homogenates both at 1 day and 14 days as measured by RL2 western blots
O-tau at S400 was ↑ at 14 days but not 1 d. pS202, pS396, pS356, and pS262 were ↓ at 1 day but not 14 d. pS400 was unchanged Chronic thiamet G ↓ pS202, pS396, pS356, pS262, and HT7 tau at 64 kD after high-speed spin (the pellet of 110,000 g × 15 min spin) but not at 50–60 kD (supernatant of the spin) Chronic thiamet G ↓ pS202/205 and AT8 tau immunoreactivity |
Graham et al. (2014) |
| Thiamet G at 10 and 500 mg/kg a single oral dose of water, 6 h | The brain-to-plasma ratio of thiamet G was shown to be < 0.1 as measured by LC-MS, ↑ 1.7× and 4 × of protein O-GlcNAcylation as measured by the quantitative sandwich immunoassay with both wheat germ agglutinin and the RL2 antibody | Hastings et al. (2017) |
| rTg4510 mice fed thiamet G at either 8 weeks or 12 weeks of age for 8 or 4 weeks duration at 3.3 mg/g in chow, achieving ∼500 mg/kg/d |
In the 8-week but not the 4-week duration group:
↓ PHF6, p-Thr Tau, and Tau aggregation without changing total Tau, as assessed by an AlphaLISA-based immunoassays in the brain insoluble fractions. ↓ CSF total Tau ↑ PNGase-resistant and β-elimination-sensitive O-tau, as assessed by click chemistry, but not RL2 or immunoprecipitation of anti-O-tau antibody 3,925 |
Hastings et al. (2017) |
|
Young mice i.p. 30 mg/kg 18 h before tMCAO
Young or old mice i.p. 30 mg/kg 30 min after onset of pMCAO |
↓ infarct | Jiang et al. (2017) |
| Young mice i.p. 20 mg/kg/d for 3 days before tMCAO or 30 min after onset of tMCAO daily till day 3 |
Neurobehavioral performance was improved
↓ infarct ↑ IL10, ↓ IL6, G-CSF, TNFα, and IL1β in both groups ↓ microglia activation as measured by Iba1+, CD16/32+, Cox-2+, iNOS+ cells |
He et al. (2017) |
| In rat spinal cord injury (SCI) model, i.v. injected within 1 h after injury at 10 mg/kg/d x 3 days |
↓ Cleaved caspase 3 at 24 h after SCI, ↓ CD68+ microglia at 7days after SCI improved neurological score at 5–20 days following SCI
↓ histological alterations of the injured spinal cord at 24 h and 21 days after SCI |
Liang et al. (2019) |
| Rats that have been injected kainate to induce seizure | Progressively↓ seizure severity after consecutive daily thiamet G injection (10 mg/kg i.p.) | Sanchez et al. (2019) |
| Single i.p. injection at 0, 10, 20, 100, 200, or 500 mg/kg | ↑ protein O-GlcNAcylation at all these doses and peaked at 20 mg/kg, 8 h after injection in the brain, liver, and knee using western blot with RL2; no change in the muscle at any of these doses | Andres-Bergos et al. (2012) |
| Male C57BL/6 mice at 23 days i.p. 20 mg/kg/d × 15 days | ↑ protein O-GlcNAcylation in the brain, liver, and muscle ↑ growth plate height and hypertrophic zone height in the endochondral plate of the tibias, suggesting a stimulation of growth plate chondrocyte differentiation |
|
| In a mouse tibialis anterior muscle injury model: i.p. 40 mg/kg 1 day after injury for 3 days | ↓ myogenin levels in tibialis anterior muscle | Kim et al., (2020) |
| STZ was injected i.p. at 50 mg/kg for 5 days, thiamet G was injected i.v. 20 mg/kg/wk 1 week after STZ for 8 weeks | ↑ vascular calcification in STZ-treated mice | Heath et al. (2014) |
| C57BL/6 mice at 8 weeks of age: i.p. 20 mg/kg/d × 15 days | ↑ protein O-GlcNAcylation in the colon as assessed by western blots using RL2 The increase of protein O-GlcNAcylation i.p. injected with thiamet G (20 mg/kg/d) for 15 days ↑ OGA and ↓ OGT at both protein and mRNA level in the colon |
Olivier-Van Stichelen et al., (2014), Decorcelle et al. (2020) |
| Single oral administration of MK-8719 to SD rats | ↑ Brain protein O-GlcNAcylation at 0.3 mg/kg; ↑ PBMC protein O-GlcNAcylation at 10 mg/kg, using a sandwich immunoassay with wheat germ agglutinin and RL2 ↑ protein O-GlcNAcylation at 30 mg/kg, 8–24 h in both brain and PBMC, with brain greater ↑ at 12 h MK-8719 levels peaked at 2 h in both the brain and plasma |
Wang et al. (2020) |
| rTg4510 mice at 8 weeks of age: Chronic diet dosing of MK-8719 at 10, 30, and 100 mg/kg | No overt adverse effects from 8 to 16 weeks of age ↓ aggregated tau after 30 mg/kg, ↓ AT8 tau after 10 mg/kg, and ↓ PHF6 tau after 30 mg/kg in insoluble tau fraction using AlphaLISA-based immunoassays |
|
| rTg4510 mice at 8 weeks of age: chronic diet dosing of MK-8719 at 100 mg/kg |
↓ brain NFT pathology using AT8 immunostaining; ↓ CSF total tau after 100 mg/kg from 8 to 20 weeks
↓ spontaneous locomotor activity in terms of distance traveled 8 weeks after of MK-8719 in these mice compared to age-matched mice with vehicle ↑ cortex and hippocampal volume were also seen 16 and 24 weeks after MK-8719 |