Abstract
The present report describes in vivo investigations of genome activity and its role in the mechanisms forming long-term synaptic plasticity in defensive behavior command neuron LPl1 during the acquisition of nociceptive sensitization by common snails. Transcription processes were recorded using SYTO 16, a specific fluorescent indicator of DNA activity, along with in vivo computer image analysis. Studies in control snails showed that application of nociceptive stimuli to the head led to biphasic changes in the bioelectrical responses of neurons to tactile and chemical stimulation-depression of responses in the short-term stage (during the 1 h after training) and their facilitation during the long-term stage of sensitization (more than 24 h). There were marked increases in fluorescence over the nucleus of the command neuron stained with SYTO 16 at 15–20 min from the start of training, this lasting 4–5 h. Acquisition of sensitization in the presence of the RNA synthesis inhibitor actinomycin D (20 μM) to the neuron led to the complete elimination of changes in fluorescence and synaptic facilitation in the responses of LPl1 to sensory stimulation in the long-term stage of sensitization but had no effect during the short-term stage of sensitization. Actinomycin D given 30 min after the end of acquisition of sensitization (1 h after the start) had no effect on the dynamics of fluorescence or synaptic facilitation. Thus, the acquisition of nociceptive sensitization is accompanied by a rapid (within 15–20 min) activation of the DNA of neuron LPl1 and subsequent (about 1 h) display of long-term synaptic facilitation. Induction of both processes was suppressed by the RNA synthesis inhibitor over a relatively short time period-1 h from the moment at which training started.
Similar content being viewed by others
REFERENCES
K. V. Anokhin, Expression of early genes in memory mechanisms,” Vestn. Ros. Akad. Nauk., 12, 58–61 (1997).
V. D. Goncharuk, S. A. Kozyrev, and V. P. Nikitin, “Acquisition of sensitization in the common snail: morphofunctional correlates in defensive behavior command neurons,” Neirofiziologiya, 1, No.2, 150–157 (1993).
O. A. Maksimova and P. M. Balaban, The Neural Mechanisms of Behavioral Plasticity [in Russian], Nauka, Moscow (1983).
A. A. Mekhtiev, S. A. Kozyrev, V. P. Nikitin, and V. V. Sherstnev, “Selective effects of antibodies to smp69 protein on the activity of defensive behavior command neurons in the common snail,” Ros. Fiziol. Zh. im. I. M. Sechenova, 89, No.4, 389–396 (2003).
V. P. Nikitin, “The transient stage of long-term synaptic facilitation in defensive behavior command neurons in sensitized snails,” Ros. Fiziol. Zh. im. I. M. Sechenova, 85, No.1, 36–47 199
V. P. Nikitin and S. A. Kozyrev, “Selective effects of the protein kinase C Inhibitor on the synaptic plasticity of defensive behavior command neurons in sensitized snails,” Ros. Fiziol. Zh. im. I. M. Sechenova, 88, No.11, 1401–1411 (2002).
V. P. Nikitin, S. A. Kozyrev, and A. V. Shevelkin, “Selective actions of opioid peptides on excitability and the various sensory inputs of defensive behavior command neurons LPl1 and RPl1 in the common snail,” Ros. Fiziol. Zh. im. I. M. Sechenova, 88, No.1, 22–31 (2002).
V. P. Nikitin, S. A. Kozyrev, and A. V. Shevelkin, “Glutamate NMDA receptor antagonists selectively influence the synaptic mechanisms of nociceptive sensitization in snails,” Zh. Vyssh. Nerv. Deyat., 50, No.4, 686–696 (2000).
A. V. Shevelkin, V. P. Nikitin, S. A. Kozyrev, and V. V. Sherstnev, “In vivo videomicroscopic studies of DNA activity using the fluorescent dye syto 16 in neurons of the common snail Helix lucorum in sensitization, ” in: Proceedings of the 18th Congress of the I. P. Pavlov Physiological Society [in Russian], Kazan’ (2001), p. 271.
A. V. Shevelkin, “An apparatus for in vivo fluorescent microscopy with a computer image analysis system, ” VNMT (2003) (in press).
C. J. Bunthof, S. van Schalkwijk, W. Meijer, T. Abee, and J. Hugenholtz, “Fluorescent method for monitoring cheese starter permeabi-lization and lysis,” Appl. Environ. Microbiol. 67, No.9, 4264–4271 (2001).
H. P. Davis and L. R. Squire, “Protein synthesis and memory. A review,” Psychol. Bull., 9, No.3, 518–559 (1984).
J. Demmer, M. Dragunow, S. E. Mason, J. D. Leah, W. C. Abraham, and W. P. Tate, “Differential expression of immediate early genes after hippocampal long-term potentiation in awake rats,” Mol. Brain Res., 17, No.3–4, 279–286 (1993).
U. Frey, S. Frey, F. Schollmeier, and M. Krug, “Influence of actinomycin D, a RNA synthesis inhibitor, on long-term potentiation in rat hippocampal neurons in vivo and in vitro,” J. Physiol. (London), 490, No.3, 703–711 (1996).
M. Ghirardi, P. G. Montarolo, and E. R. Kandel, “A novel intermediate stage in the transition between short-and long-term facilitation in the sensory to motor neuron synapse of Aplysia,” Neuron, 14, No.2, 413–420 (1995).
P. Goelet, V. F. Castellucci, S. Schacher, and E. Kandel, “The long and the short of long-term memory — a molecular framework,” Nature, 322, No.6078, 419–422 (1986).
Y. Y. Huang, X. C. Li, and E. R. Kandel, “cAMP contributes to mossy fiber LTP by initiating both a covalently mediated early phase and macromolecular synthesis-dependent late phase,” Cell, 79, No.1, 69–79 (1994).
R. P. Hougland, Handbook of Fluorescent Probes and Research Chemicals, Molecular Probes Inc. (2001), 8th Edition.
R. P. Hougland, Personal communication (2002).
B. S. Kauderer and E. R. Kandel, “Capture of a protein synthesis-dependent component of long-term depression,” Proc. Natl. Acad. Sci. USA, 97, No.5, 2253–2258 (2000).
B. Khoobehi and G. A. Peyman, “Fluorescent labeling of blood cells for evaluation of retinal and choroidal circulation,” Ophthalmic Surg. Lasers, 30, No.2, 140–145 (1999).
D. Manahan-Vaughan, A. Kulla, and J. U. Frey, “Requirement of translation but not transcription for the maintenance of long-term depression in the CA1 region of freely moving rats,” J. Neurosci., 20, No.22, 8572–8576 (2000).
B. Milner, L. R. Squire, and E. R. Kandel, “Cognitive neuroscience and the study of memory,” Neuron, 20, No.3, 445–468 (1998).
P. G. Montarolo, P. Goelet, V. F. Castellucci, J. Morgan, E. R. Kandel, and S. A. Schacher, “A critical period for macromolecular synthesis in long-term heterosynaptic facilitation in Aplysiaa,” Science, 234, No.4781, 1249–1254 (1986).
P. V. Nguyen, T. Abel, and E. R. Kandel, “Requirement of a critical period of transcription for induction of a late phase of LTP,” Science, 265, No.5175, 1104–1107 (1994).
T. J. Nelson and D. L. Alkon, “Protein changes underlying long-term facilitation in Aplysia, ” Bioassays, 11, No.4, 106–108 (1989).
F. Noel, K. P. Scholz, A. Eskin, and J. H. Byrne, “Common set of proteins in Aplysia sensory neurons affected by an in vitro analogue of long-term sensitization training, 5-HT, and cAMP,” Brain Res., 568, No.1–2, 67–75 (1991).
S. Otani, C. J. Marchall, W. P. Tate, G. V. Goddard, and W. C. Abraham, “Maintenance of long-term potentiation in rat dentate gyrus requires protein synthesis but not messenger RNA synthesis immediately post-tetanization,” Neurosci., 28, No.3, 519–526 (1989).
O. Steward and P. Worley, “A cellular mechanism for targeting newly synthesized mRNAs to synaptic sites on dendrites,” Proc. Natl. Acad. Sci. USA, 98, No.13, 7062–7068 (2001).
Author information
Authors and Affiliations
Additional information
Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 90, No. 2, pp. 157–168, February, 2004.
Rights and permissions
About this article
Cite this article
Shevelkin, A.V., Kozyrev, S.A., Nikitin, V.P. et al. In Vivo Investigation of Genome Activity and Synaptic Plasticity of Neurons in Snails During Learning. Neurosci Behav Physiol 35, 595–603 (2005). https://doi.org/10.1007/s11055-005-0099-9
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s11055-005-0099-9