Polymerization of Tubulin and Its Colchicine Binding Activity in Various Brain Structures in Healthy and in Schizophrenia

Introduction: abnormalities of neuronal cytoskeleton in mental disorders require to study microtubules and their proteins. Tubulin (the main protein of microtubules) has specific properties: reversibly polymerize into microtubules and bind mitotic poison colchicine in equimolar quantities.

Objective: to evaluate the process of tubulin polymerization by light scattering change and to determine the level of colchicine binding (colchicine binding activity of tubulin) in various brain structures in healthy and schizophrenia brains.

Material and methods: autopsy brain samples from patients with schizophrenia (n = 6) and from the control group (n = 9) were studied. Samples of the prefrontal (area 10), temporal (area 21), cingulate cortex (area 23/24) and thalamus were isolated (Brodmann's areas). Measurements of light scattering during tubulin polymerization and colchicine-binding activity of tubulin were determined as described earlier.

Results: tubulin polymerization was not disturbed in schizophrenia as compared to controls, except for the cingulate cortex that showed slight but significant decrease in light scattering. At the same time, the binding of colchicine in schizophrenia was reduced in all examined areas of the cortex. This decrease was not associated with age, sex, and postmortem interval since the groups were matched by these factors. The tubulin colchicine-binding activity in thalamus remained at the same level both in control and schizophrenia, but it was lower than in the areas of the cortex.

Conclusion: Decreased activity and hence decreased amount of tubulin in the cerebral cortical areas without changes of the tubulin polymerization in microtubules have been shown in schizophrenia. The results confirm the literature data on the changes in the cytoskeleton in cortical areas in schizophrenia.

1. Pchitskaya EI, Zhemkov VA, Bezprozvanny IB. Dynamic microtubules in Alzheimer's disease: association with dendritic spine pathology. Biochemistry. 2018;83(9): 1343-1350. (In Russ.). DOI: 10.1134/S0006297918090080.

2. Marchisella F, Coffey ET, Hollos P. Microtubule and microtubule associated protein anomalies in psychiatric disease. Cytoskeleton (Hoboken). 2016;73(10):596-611. DOI: 10.1002/cm.21300.

3. Behan AT, Byrne C, Dunn MJ, et al. Proteomic analysis of membrane microdomain-associated proteins in the dorsolateral prefrontal cortex in schizophrenia and bipolar disorder reveals alterations in LAMP, STXBP1 and BASP1 protein expression. Mol. Psychiatry. 2009;14(6):601-613. DOI: 10.1038/mp.2008.7

4. Jaworski J, Kapitein LC, Gouveia SM, et al. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron. 2009;61(1):85-100. DOI: 10.1016/j.neuron.2008.11.013.

5. Gu J, Zheng JQ. Microtubules in Dendritic Spine Development and Plasticity. Open Neurosci. J. 2009;3:128-133. DOI: 10.2174/1874082000903020128.

6. Martins-de-Souza D, Schmitt A, Roder R, et al. Sex-specific proteome differences in the anterior cingulate cortex of schizophrenia. J. Psychiatr. Res. 2010;44(14):989-991. DOI: 10.1016/j.jpsychires.2010.03.003.

7. Verstraelen P, Detrez JR, Verschuuren M, et al. Dysregulation of Microtubule Stability Impairs Morphofunctional Connectivity in Primary Neuronal Networks. Front Cell. Neurosci. 2017;11:173. DOI: 10.3389/fncel.2017.00173.

8. Andrieux A, Salin P, Schweitzer A, et al. Microtubule stabilizer ameliorates synaptic function and behavior in a mouse model for schizophrenia. Biol. Psychiatry. 2006;60(11):1224-1230. DOI: 10.1016/j.biopsych.2006.03.048.

9. Gardiner J, Overall R, Marc J. The microtubule cytoskeleton acts as a key downstream effector of neurotransmitter signaling. Synapse. 2011;65(3):249-256. DOI:10.1002/syn.20841.

10. Janke C. The tubulin code: Molecular components, readout mechanisms, and functions. J. Cell. Biol. 2014;l4:461-472. DOI:10,1083/jcb.201406055.

11. Androsova LV, Burbaeva GSh. Aminazin — vysokoeffektivnyj ingibitor polimerizacii tubulina. Biohimiya. 1987;52(7):1162-1167. (In Russ).

12. Shevtsov PN, Shevtsova EF, Burbaeva GSh. Effect of Aluminum, Iron, and Zinc Ions on the Assembly of Microtubules from Brain Microtubule Proteins. Bull. Exp. Biol. Med. 2016;161(4):451-455. (In Russ). DOI: 10.1007/s10517-016-3436-9.

13. Shevtsov PN, Shevtsova EF, Savushkina OK i dr. Influence of Al3+, Fe3+ и Zn2 ions on phosphorylation of tubulin and microtubulo-associated proteins of rat brain. Bull. Exp. Biol. Med. 2018;165(4):512-515. (In Russ). DOI: 10.1007/s10517-018-4206-7.

14. Bauer DE, Haroutunian V, McCullumsmith RE, Meador-Woodruff JH. Expression of four housekeeping proteins in elderly patients with schizophrenia. J. Neural. Transm. (Vienna). 2009;116(4):487-491. DOI: 10.1007/s00702-008-0143-3.

15. Sivagnanasundaram S, Crossett B, Dedova I, et al. Abnormal pathways in the genu of the corpus callosum in schizophrenia pathogenesis: a proteome study. Proteomics Clin. Appl. 2007;1(10):1291-1305. DOI: 10.1002/prca.200700230.

16. Chan MK, Tsang TM, Harris LW, et al. Evidence for disease and antipsychotic medication effects in post-mortem brain from schizophrenia patients. Mol. Psychiatry. 2011;16(12):1189-1202. DOI: 10.1038/mp.2010.100.

17. Clinton SM, Haroutunian V, Meador-Woodruff JH. Up-regulation of NMDA receptor subunit and postsynaptic density protein expression in the thalamus of elderly patients with schizophrenia. J. Neurochem. 2006;98(4):1114-1125. DOI: 10.1111/j.1471-4159.2006.03954.x.

18. Beasley CL, Pennington K, Behan A, et al. Proteomic analysis of the anterior cingulate cortex in the major psychiatric disorders: Evidence for disease-associated changes. Proteomics. 2006;6(11):3414-3425. DOI: 10.1002/pmic.200500069.

19. English JA, Dicker P, Focking M, et al. 2-D DIGE analysis implicates cytoskeletal abnormalities in psychiatric disease. Proteomics. 2009;9(12):3368 -3382. DOI: 10.1002/pmic.200900015.

20. Hayashi-Takagi A, Takaki M, Graziane N, et al. Disrupted-in-Schizophrenia 1 (DISC1) regulates spines of the glutamate synapse via Rac1. Nat. Neurosci. 2010;13(3):327-332. DOI: 10.1038/nn.2487.