Влияние LIM-киназы 1 на память и брачную песню cамцов дрозофилы зависит от типа нейронов

Сигнальный каскад ремоделирования актина, в состав которого входят LIM-киназа 1 (LIMK1) и ее субстрат кофилин, участвует в регуляции различных процессов в нейронах позвоночных и беспозвоночных животных. Drosophila melanogaster широко используется как модельный объект для изучения механизмов формирования, сохранения и воспроизведения памяти, а также забывания. Ранее активное забывание у дрозофилы исследовали с помощью классического павловского ольфакторного обучения. Было показано, что в разных формах забывания участвуют специфические дофаминергические нейроны и компоненты актинового каскада. В данной работе мы оценивали роль LIMK1 в процессах памяти и забывания у дрозофилы в парадигме условно-рефлекторного подавления ухаживания. В мозге дрозофилы уровень LIMK1 и фосфокофилина избирательно снижен в отдельных структурах нейропиля, включая лопасти грибовидных тел и центральный комплекс. В то же время LIMK1 присутствует в телах нервных клеток, таких как кластеры дофаминергических нейронов, регулирующие формирование памяти при условно-рефлекторном подавлении ухаживания. С использованием системы бинарного скрещивания GAL4 × UAS мы инициировали РНК-интерференцию limk1 в различных типах нервных клеток. У гибридных линий с интерференцией limk1 в лопастях грибовидных тел и глии наблюдалось усиление 3-часовой краткосрочной памяти, без видимого влияния на долгосрочную память. Интерференция limk1 в холинергических нейронах приводила к снижению краткосрочной памяти, в дофаминергических и серотонинергических нейронах ее результатом было также существенное нарушение способности мух к обучению. Напротив, интерференция limk1 в нейронах fruitless усиливала 15–60-минутную краткосрочную память, что указывает на возможную роль LIMK1 в процессах активного забывания. У самцов с интерференцией limk1 в  холинергических и fruitless нейронах также были отмечены разнонаправленные изменения параметров брачной песни. Таким образом, эффекты LIMK1 на память и брачную песню самцов дрозофилы определяются типом нервных клеток или структурой мозга.

1. Abe H., Nagaoka R., Obinata T. Cytoplasmic localization and nuclear transport of cofilin in cultured myotubes. Exp. Cell Res. 1993; 206(1):1-10. DOI 10.1006/excr.1993.1113.

2. Abe T., Yamazaki D., Murakami S., Hiroi M., Nitta Y., Maeyama Y., Tabata T. The NAV2 homolog Sickie regulates F-actin-mediated axonal growth in Drosophila mushroom body neurons via the noncanonical Rac-Cofilin pathway. Development. 2014;141(24):47164728. DOI 10.1242/dev.113308.

3. Albin S.D., Kaun K.R., Knapp J.M., Chung P., Heberlein U., Simpson J.H. A subset of serotonergic neurons evokes hunger in adult Drosophila. Curr. Biol. 2015;25(18):2435-2440. DOI 10.1016/j.cub.2015.08.005.

4. Aso Y., Grübel K., Busch S., Friedrich A.B., Siwanowicz I., Tanimoto H. The mushroom body of adult Drosophila characterized by GAL4 drivers. J. Neurogenet. 2009;23(1-2):156-172. DOI 10.1080/01677060802471718.

5. Aso Y., Hattori D., Yu Y., Johnston R.M., Iyer N.A., Ngo T.B., Dionne H., Abbott L.F., Axel R., Tanimoto H., Rubin G.M. The neuronal architecture of the mushroom body provides a logic for associative learning. eLife. 2014a;3:e04577. DOI 10.7554/eLife.04577.

6. Aso Y., Sitaraman D., Ichinose T., Kaun K.R., Vogt K., Belliart-Guérin G., Plaçais P., Robie A.A., Yamagata N., Schnaitmann C., Rowell W.J., Johnston R.M., Ngo T.B., Chen N., Korff W., Nitabach M.N., Heberlein U., Preat T., Branson K.M., Tanimoto H., Rubin G.M. Mushroom body output neurons encode valence and guide memory-based action selection in Drosophila. eLife. 2014b; 3:e04580. DOI 10.7554/eLife.04580.

7. Barnstedt O., Owald D., Felsenberg J., Brain R., Moszynski J.P., Talbot C.B., Perrat P.N., Waddell S. Memory-relevant mushroom body output synapses are cholinergic. Neuron. 2016;89(6):1237-1247. DOI 10.1016/j.neuron.2016.02.015.

8. Ben Zablah Y., Zhang H., Gugustea R., Jia Z. LIM-kinases in synaptic plasticity, memory, and brain diseases. Cells. 2021;10(8):20792103. DOI 10.3390/cells10082079.

9. Berry J.A., Cervantes-Sandoval I., Nicholas E.P., Davis R.L. Dopamine is required for learning and forgetting in Drosophila. Neuron. 2012;74(3):530-542. DOI 10.1016/j.neuron.2012.04.007.

10. Berry J.A., Phan A., Davis R.L. Dopamine neurons mediate learning and forgetting through bidirectional modulation of a memory trace. Cell Rep. 2018;25(3):651-662. DOI 10.1016/j.celrep.2018.09.051.

11. Cervantes-Sandoval I., Chakraborty M., MacMullen C., Davis R.L. Scribble scaffolds a signalosome for active forgetting. Neuron. 2016;90(6):1230-1242. DOI 10.1016/j.neuron.2016.05.010.

12. Clyne J.D., Miesenböck G. Sex-specific control and tuning of the pattern generator for courtship song in Drosophila. Cell. 2008;133(2): 354-363. DOI 10.1016/j.cell.2008.01.050.

13. Cook K.R., Parks A.L., Jacobus L.M., Kaufman T.C., Matthews K.A. New research resources at the Bloomington Drosophila Stock Center. Fly (Austin). 2010;4(1):88-91. DOI 10.4161/fly.4.1.11230.

14. Cuberos H., Vallée B., Vourc’h P., Tastet J., Andres C.R., Bénédetti H. Roles of LIM kinases in central nervous system function and dysfunction. FEBS Lett. 2015;589(24 Pt.B):3795-3806. DOI 10.1016/j.febslet.2015.10.032.

15. Davis R.L., Zhong Y. The biology of forgetting – a perspective. Neuron. 2017;95(3):490-503. DOI 10.1016/j.neuron.2017.05.039.

16. Duffy J.B. GAL4 system in Drosophila: a fly geneticist’s Swiss army knife. Genesis. 2002;34(1-2):1-15. DOI 10.1002/gene.10150.

17. Gao Y., Shuai Y., Zhang X., Peng Y., Wang L., He J., Zhong Y., Li Q. Genetic dissection of active forgetting in labile and consolidated memories in Drosophila. Proc. Natl. Acad. Sci. USA. 2019;116(42): 21191-21197. DOI 10.1073/pnas.1903763116.

18. Hartenstein V. Morphological diversity and development of glia in Drosophila. Glia. 2011;59(9):1237-1252. DOI 10.1002/glia.21162.

19. Iliadi K.G., Kamyshev N.G., Popov A.V., Iliadi N.N., Rashkovetskaya E.L., Nevo E., Korol A.B. Peculiarities of the courtship song in the Drosophila melanogaster populations adapted to gradient of microecological conditions. J. Evol. Biochem. Physiol. 2009;45(5): 579-588. DOI 10.1134/S0022093009050041.

20. Jones S.G., Nixon K.C.J., Chubak M.C., Kramer J.M. Mushroom body specific transcriptome analysis reveals dynamic regulation of learning and memory genes after acquisition of long-term courtship memory in Drosophila. G3: Genes Genomes Genetics (Bethesda). 2018;8(11):3433-3446. DOI 10.1534/g3.118.200560.

21. Kahsai L., Winther A.M. Chemical neuroanatomy of the Drosophila central complex: distribution of multiple neuropeptides in relation to neurotransmitters. J. Comp. Neurol. 2011;519(2):290-315. DOI 10.1002/cne.22520.

22. Kamyshev N.G., Iliadi K.G., Bragina J.V. Drosophila conditioned courtship: two ways of testing memory. Learn. Mem. 1999;6(1): 1-20.

23. Kasture A.S., Hummel T., Sucic S., Freissmuth M. Big lessons from tiny flies: Drosophila melanogaster as a model to explore dysfunction of dopaminergic and serotonergic neurotransmitter systems. Int. J. Mol. Sci. 2018;19(6):1788-1805. DOI 10.3390/ijms19061788.

24. Keleman K., Vrontou E., Krüttner S., Yu J.Y., Kurtovic-Kozaric A., Dickson B.J. Dopamine neurons modulate pheromone responses in Drosophila courtship learning. Nature. 2012;489(7414):145-149. DOI 10.1038/nature11345.

25. Krüttner S., Traunmüller L., Dag U., Jandrasits K., Stepien B., Iyer N., Fradkin L.G., Noordermeer J.N., Mensh B.D., Keleman K. Synaptic Orb2A bridges memory acquisition and late memory consolidation in Drosophila. Cell Rep. 2015;11(12):1953-1965. DOI 10.1016/j.celrep.2015.05.037.

26. Liu W., Ganguly A., Huang J., Wang Y., Ni J.D., Gurav A.S., Aguilar M.A., Montell C. Neuropeptide F regulates court-ship in Drosophila through a male-specific neuronal circuit. eLife. 2019;8:1-29. DOI 10.7554/eLife.49574.

27. Lopatina N.G., Zachepilo T.G., Chesnokova E.G., Savateeva-Popova E.V. Mutations in structural genes of tryptophan metabolic enzymes of the kynurenine pathway modulate some units of the L-glutamate receptor – actin cytoskeleton signaling cascade. Russ. J. Genet. 2007;43(10):1168-1172. DOI 10.1134/S1022795407100110.

28. Mao Z., Davis R.L. Eight different types of dopaminergic neurons innervate the Drosophila mushroom body neuropil: anatomical and physiological heterogeneity. Front. Neural Circuits. 2009;3:1-17. DOI 10.3389/neuro.04.005.2009.

29. Mazzoni D., Dannenberg R. Audacity®. Audacity Team, 2020. URL: https://audacityteam.org. Last access: 05.12.2021.

30. Medina J.H. Neural, cellular and molecular mechanisms of active forgetting. Front. Syst. Neurosci. 2018;12:1-10. DOI 10.3389/fnsys.2018.00003.

31. Medvedeva A.V., Molotkov D.A., Nikitina E.A., Popov A.V., Karagodin D.A., Baricheva E.M., Savvateeva-Popova E.V. Systemic regulation of genetic and cytogenetic processes by a signal cascade of actin remodeling: locus agnostic in Drosophila. Russ. J. Genet. 2008; 44(6):669-681. DOI 10.1134/S1022795408060069.

32. Medvedeva A.V., Tokmatcheva E.V., Kaminskaya A.N., Vasileva S.A., Nikitina E.A., Zhuravlev A.V., Zakharov G.A., Zatsepina O.G., Savvateeva-Popova E.V. Parent-of-origin effects on nuclear chromatin organization and behavior in a Drosophila model for Williams–Beuren Syndrome. Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2021;25(5):472-485. DOI 10.18699/VJ21.054.

33. Montague S.A., Baker B.S. Memory elicited by courtship conditioning requires mushroom body neuronal subsets similar to those utilized in appetitive memory. PLoS One. 2016;11(10):e0164516. DOI 10.1371/journal.pone.0164516.

34. Ng J., Luo L. Rho GTPases regulate axon growth through convergent and divergent signaling pathways. Neuron. 2004;44(5):779-793. DOI 10.1016/j.neuron.2004.11.014.

35. Nikitina E.A., Medvedeva A.V., Zakharov G.A., Savvateeva-Popova E.V. The Drosophila agnostic locus: involvement in the formation of cognitive defects in Williams syndrome. Acta Naturae. 2014; 6(2):53-61. DOI 10.32607/20758251-2014-6-2-53-61.

36. Nikitina E.A., Zhuravlev A.V., Savvateeva-Popova E.V. Effect of impaired kynurenine synthesis on memory in Drosophila. Integrativnyaya Fiziologiya = Integrative Physiology. 2021;2(1):49-60. DOI 10.33910/2687-1270-2021-2-1-49-60. (in Russian)

37. Okamoto N., Nishimura T. Signaling from glia and cholinergic neurons controls nutrient-dependent production of an insulin-like peptide for Drosophila body growth. Dev. Cell. 2015;35(3):295-310. DOI 10.1016/j.devcel.2015.10.003.

38. Popov A.V., Peresleni A.I., Savvateeva-Popova E.V., Vol’f R., Heisenberg M. The role of the mushroom bodies and of the central complex of Drosophila melanogaster brain in the organization of courtship behavior and communicative sound production. J. Evol. Biochem. Phys. 2004;40:641-652. DOI 10.1007/s10893-004-0005-z.

39. Redt-Clouet C., Trannoy S., Boulanger A., Tokmatcheva E., Savvateeva-Popova E., Parmentier M.L., Preat T., Dura J.M. Mushroom body neuronal remodelling is necessary for short-term but not for long-term courtship memory in Drosophila. Eur. J. Neurosci. 2012; 35(11):1684-1691. DOI 10.1111/j.1460-9568.2012.08103.x.

40. Ritchie M.G., Yate V.H., Kyriacou C.P. Genetic variability of the interpulse interval of courtship song among some European populations of Drosophila melanogaster. Heredity (Edinb). 1994;72(Pt.5):459464. DOI 10.1038/hdy.1994.64.

41. Salvaterra P.M., Kitamoto T. Drosophila cholinergic neurons and processes visualized with Gal4/UAS-GFP. Brain Res. Gene Expr. Patterns. 2001;1(1):73-82. DOI 10.1016/s1567-133x(01)00011-4.

42. Savvateeva-Popova E.V., Popov A.V., Grossman A., Nikitina E.A., Medvedeva A.V., Peresleni A.I., Molotkov D.A., Kamyshev N.G., Pyatkov K.I., Zatsepina O.G., Schostak N., Zelentsova E.S., Pavlova G., Panteleev D., Riederer P., Evgen’ev M.B. Non-coding RNA as a trigger of neuropathologic disorder phenotypes in transgenic Drosophila. J. Neural Transm. (Vienna). 2008;115(12):1629-1642. DOI 10.1007/s00702-008-0078-8.

43. Savvateeva-Popova E.V., Popov A.V., Heinemann T., Riederer P. Drosophila mutants of the kynurenine pathway as a model for ageing studies. Adv. Exp. Med. Biol. 2003;527:713-722. DOI 10.1007/9781-4615-0135-0_84.

44. Savvateeva-Popova E.V., Zhuravlev A.V., Brázda V., Zakharov G.A., Kaminskaya A.N., Medvedeva A.V., Nikitina E.A., Tokmatcheva E.V., Dolgaya J.F., Kulikova D.A., Zatsepina O.G., Funikov S.Y., Ryazansky S.S., Evgen’ev M.B. Drosophila model for the analysis of genesis of LIM-kinase 1-dependent Williams–Beuren syndrome cognitive phenotypes: INDELs, transposable elements of the Tc1/ mariner superfamily and microRNAs. Front. Genet. 2017;8:123. DOI 10.3389/fgene.2017.00123.

45. Shuai Y., Lu B., Hu Y., Wang L., Sun K., Zhong Y. Forgetting is regulated through Rac activity in Drosophila. Cell. 2010;140(4):579589. DOI 10.1016/j.cell.2009.12.044.

46. Shuai Y., Zhong Y. Forgetting and small G protein Rac. Protein Cell. 2010;1(6):503-506. DOI 10.1007/s13238-010-0077-z.

47. Siegel R.W., Hall J.C. Conditioned responses in courtship behavior of normal and mutant Drosophila. Proc. Natl. Acad. Sci. USA. 1979; 76(7):3430-3434. DOI 10.1073/pnas.76.7.343.

48. Strauss R., Heisenberg M. A higher control center of locomotor behavior in the Drosophila brain. J. Neurosci. 1993;13(5):1852-1861. DOI 10.1523/JNEUROSCI.13-05-01852.1993.

49. Sun B., Xu P., Salvaterra P.M. Dynamic visualization of nervous system in live Drosophila. Proc. Natl. Acad. Sci. USA. 1999;96(18): 10438-10443. DOI 10.1073/pnas.96.18.10438.

50. Thapa A., Sullivan S.M., Nguyen M.Q., Buckley D., Ngo V.T., Dada A.O., Blankinship E., Cloud V., Mohan R.D. Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system. Biotechniques. 2019;67(6):299-305. DOI 10.2144/btn-2018-0067.

51. Tully T., Preat T., Boynton S.C., Vecchio M.D. Genetic dissection of consolidated memory in Drosophila. Cell. 1994;79(1):35-47. DOI 10.1016/0092-8674(94)90398-0.

52. Virtual Fly Brain. URL: https://v2.virtualflybrain.org. Last access: 01.12.2021.

53. von Philipsborn A.C., Liu T., Yu J.Y., Masser C., Bidaye S.S., Dickson B.J. Neuronal control of Drosophila courtship song. Neuron. 2011;69(3):509-522. DOI 10.1016/j.neuron.2011.01.011.

54. Winbush A., Reed D., Chang P.L., Nuzhdin S.V., Lyons L.C., Arbeitman M.N. Identification of gene expression changes associated with long-term memory of courtship rejection in Drosophila males. G3: Genes Genomes Genetics (Bethesda). 2012;2(11):1437-1445. DOI 10.1534/g3.112.004119.

55. Xiao C., Qiu S., Robertson R.M. The white gene controls copulation success in Drosophila melanogaster. Sci. Rep. 2017;7(1):77127725. DOI 10.1038/s41598-017-08155-y.

56. Yasuyama K., Meinertzhagen I.A., Schürmann F.W. Synaptic organization of the mushroom body calyx in Drosophila melanogaster. J. Comp. Neurol. 2002;445(3):211-226. DOI 10.1002/cne.10155.

57. Yi W., Zhang Y., Tian Y., Guo J., Li Y., Guo A. A subset of cholinergic mushroom body neurons requires Go signaling to regulate sleep in Drosophila. Sleep. 2013;36:1809-1821. DOI 10.5665/sleep.3206.

58. Yu J.Y., Kanai M.I., Demir E., Jefferis G.S., Dickson B.J. Cellular organization of the neural circuit that drives Drosophila courtship behavior. Curr. Biol. 2010;20(18):1602-1614. DOI 10.1016/j.cub.2010.08.025.

59. Zalomaeva E.S., Falina V.S., Medvedeva A.V., Nikitina E.A., Savvateeva-Popova E.V. Learning and forgetting in Drosophila melanogaster in limk1 gene polymorphism. Integrativnyaya Fiziologiya = Integrative Physiology. 2021;2(3):318-327. DOI 10.33910/26871270-2021-2-3-318-327. (in Russian)

60. Zhang X., Li Q., Wang L., Liu Z.J., Zhong Y. Cdc42-dependent forgetting regulates repetition effect in prolonging memory retention. Cell Rep. 2016;16(3):817-825. DOI 10.1016/j.celrep.2016.06.041.

61. Zhao X., Lenek D., Dag U., Dickson B.J., Keleman K. Persistent activity in a recurrent circuit underlies courtship memory in Drosophila. eLife. 2018;7:1-16. DOI 10.7554/eLife.31425.

62. Zhuravlev A.V., Shchegolev B.F., Zakharov G.A., Ivanova P.N., Nikitina E.A., Savvateeva-Popova E.V. 3-Hydroxykynurenine as a potential ligand for Hsp70 proteins and its effects on Drosophila memory after heat shock. Mol. Neurobiol. 2022;59(3):1862-1871. DOI 10.1007/s12035-021-02704-3.

63. Zhuravlev A.V., Vetrovoy O.V., Ivanova P.N., Savvateeva-Popova E.V. 3-Hydroxykynurenine in regulation of Drosophila behavior: the novel mechanisms for cardinal phenotype manifestations. Front. Physiol. 2020;11:971. DOI 10.3389/fphys.2020.00971.