, Volume 40, Issue 4, pp 295–303 | Cite as

Fos Immunoreactivity and NADPH-d Reactivity in the Brain Cortex of Rats Realizing Motivated Stereotyped Movements by the Forelimb

  • O. V. Vlasenko
  • A. I. Pilyavskii
  • V. A. Maiskii
  • A. V. Maznichenko

A comparative study of mmunoreactivity with respect to c-Fos protein in the motor (zones М1 and М2), medial prefrontal (PrL and IL), and cingular (Cg1 and Cg2) cortices allowed us to find significant differences between the intensities of expression of gene c-fos in these cortical regions in control rats (group 1) and animals trained to perform catching of food globules by the forelimb (i.e., realizing an operant food-procuring reflex, group 2). The density of distribution of Fos-immunoreactive (Fos-ir) neurons in rats of group 2 in motor and limbic cortical zones at +2.2 to +0.2 levels rostrally from the bregma were significantly lower than in control rats (Р < 0.05). In animals of group 2, we also found significantly greater numbers of Fos-ir neurons in the contralateral (with respect to the active extremity) zones of the cortex at all examined levels. These changes are probably related to functional changes in the cortex resulting from learning of motor habits in the course of training sessions for stabilization of the operant reflex. Histochemical estimation of the NADPH-diaphorase (NADPH-d) activity in the motor and limbic cortex showed that, in rats of both groups, the maximum number of labeled interneurons per slice in the М1 zone were observed in layers II/III, V, and VI (5.6 ± 0.4, 6.4 ± 0.5, and 14.0 ± 0.8, respectively, within 200 × 200 μm2 areas). In the limbic cortex, NADPH-d-reactive (NADPH-d-r) interneurons were also met in layers II/III, V, and VI. Cortical NADPHd-r neurons with the Fos-ir nuclei were not found. The presence of spatial associations of the somata or processes of NADPH-d-r neurons with intraparenchimal arterioles and microvessels was a typical feature of the distribution of NADPH-d-reactivity in the М1 and М2 zones, as well as in Cg1, Cg2, PrL, and IL. The following succession of the density of neurovascular associations was observed: Cg1 Cg2 М1 М2 > > PrL. As is supposed, NADPH-d-r neurons (i.e., cells generating NO) are involved in the control of regional blood flow in the studied cortical regions.


c-fos expression NADPH-diaphorase activity operant reflex food-related motivation motor cortex limbic cortex 


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  1. 1.
    A. Canedo, “Primary motor cortex influences on the descending and ascending systems,” Prog. Neurobiol., 51, 287–335 (1997).PubMedCrossRefGoogle Scholar
  2. 2.
    T. M. Tzschentke and W. J. Schmidt, “Functional relationship among medial prefrontal cortex, nucleus accumbens, and ventral tegmental area in locomotion and reward,” Crit. Rev. Neurobiol., 14, 131–142 (2000).PubMedGoogle Scholar
  3. 3.
    P. Hlustik, A. Solodkin, R. P. Gullapalli, et al., “Somatotopy in human primary motor and somatosensory hand representations revisited,” Cerebr. Cortex, 11, 312–321 (2001).CrossRefGoogle Scholar
  4. 4.
    H. Alkadhi, G. R. Crelier, S. H. Boendermaker, et al., “Somatotopy in the ipsilateral primary motor cortex,” NeuroReport, 13, 2065–2070 (2002).PubMedCrossRefGoogle Scholar
  5. 5.
    C. Stippich, M. Blatow, A. Durst, et al., “Global activation of primary motor cortex during voluntary movements in man,” Neuroimage, 34, 1227–1237 (2007).PubMedCrossRefGoogle Scholar
  6. 6.
    J. N. Sanes and J. P. Donoghue, “Plasticity and primary motor cortex,” Annu. Rev. Neurosci., 23, 393–415 (2000).PubMedCrossRefGoogle Scholar
  7. 7.
    D. V. Buonomano and M. M. Merzenich, “Cortical plasticity: from synapses to maps,” Annu. Rev. Neurosci., 21, 149–186 (1998).PubMedCrossRefGoogle Scholar
  8. 8.
    S. P. Hunt, A. Pini, and G. Evan, “Induction of c-foslike protein in spinal cord neurons following sensory stimulation,” Nature, 328, 632–634 (1987).PubMedCrossRefGoogle Scholar
  9. 9.
    O. V. Vlasenko, V. A. Maiskii, A. V. Maznichenko, et al., “Study of c-fos expression and NADPH-diaphorase activity in the spinal cord and brain after the development of fatigue in the neck muscles in rats,” Fiziol. Zh., 52, No. 1, 3–14 (2006).PubMedGoogle Scholar
  10. 10.
    J. A. Kleim, E. Lussnig, E. R. Schwarz, et al., “Synaptogenesis and Fos expression in the motor cortex of the adult rat after motor skill learning,” J. Neurosci., 16, 4529–4535 (1996).PubMedGoogle Scholar
  11. 11.
    M. H. Monfils, E. J. Plautz, and J. A. Kleim, “In search of the motor engram: motor map plasticity as a mechanism for encoding motor experience,” Neuroscientist, 11, 471–483 (2005).PubMedCrossRefGoogle Scholar
  12. 12.
    C. H. Liu, Y. R. Kim, J. Q. Ren, et al., “Imaging cerebral gene transcripts in live animals,” J. Neurosci., 27, 713–722 (2007).PubMedCrossRefGoogle Scholar
  13. 13.
    T. Herdegen and J. D. Leah, “Inducible and constitutive transcription factors in the mammalian nervous system: control of gene expression by Jun, Fos and Krox, and CREB/ATF proteins,” Brain Res.–Brain Res. Rev., 28, 370–490 (1998).PubMedCrossRefGoogle Scholar
  14. 14.
    R. E. Coggeshall, “Fos, nociception and the dorsal horn,” Prog. Neurobiol., 77, 299–352 (2005).PubMedGoogle Scholar
  15. 15.
    V. Bertaina-Anglade, G. Tramu, and C. Destrade, “Differential learning-stage dependent patterns of c-Fos protein expression in brain regions during the acquisition and memory consolidation of an operant task in mice,” Eur. J. Neurosci., 12, 3803–3812 (2000).PubMedCrossRefGoogle Scholar
  16. 16.
    J. L. Neisewander, D. A. Baker, R. A. Fuchs, et al., “Fos protein expression and cocaine-seeking behavior in rats after exposure to a cocaine self-administration environment,” J. Neurosci., 20, 798–805 (2000).PubMedGoogle Scholar
  17. 17.
    O. E. Svarnik, Y. I. Alexandrov, V. V. Gavrilov, et al., “Fos expression and task-related neuronal activity in rat cerebral cortex after instrumental learning,” Neuroscience, 136, 33–42 (2005).PubMedCrossRefGoogle Scholar
  18. 18.
    S.-M. Hsu, L. Raine, and H. Fanger, “Use of avidinbiotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures,” J. Histochem. Cytochem., 29, 577–580 (1981).PubMedGoogle Scholar
  19. 19.
    A. I. Pilyavskii, A. V. Maznychenko, V. A. Maisky, et al., “Capsaicin-induced effects on c-fos expression and NADPH-diaphorase activity in the feline spinal cord,” Eur. J. Pharmacol., 521, 70–78 (2005).PubMedCrossRefGoogle Scholar
  20. 20.
    G. Paxinos and C. Watson, The Rat Brain in Stereotaxic Coordinates, Academic Press, San Diego (1997).Google Scholar
  21. 21.
    S. R. Vincent and H. Kimura, “Histochemical mapping of nitric oxide synthase in the rat brain,” Neuroscience, 46, 755–784 (1992).PubMedCrossRefGoogle Scholar
  22. 22.
    C. A. Schiltz, Q. Z. Bremer, C. F. Landry, and A. E. Kelley, “Food-associated cues alter forebrain functional connectivity as assessed with immediate early gene and proenkephalin expression,” BioMed Centr. Biol., 5, 16 (2007).Google Scholar
  23. 23.
    R. D. Hall and E. P. Lindholm, “Organization of motor and somatosensory neocortex in the albino rat,” Brain Res., 66, 23–38 (1974).CrossRefGoogle Scholar
  24. 24.
    U. Ziemann, T. V. Ilić, C. Pauli, et al., “Learning modifies subsequent induction of long-term potentiation-like and long-term depression-like plasticity in human motor cortex,” J. Neurosci., 24, 1666–1672 (2004).PubMedCrossRefGoogle Scholar
  25. 25.
    G. Q. Bi and M. M. Poo, “Synaptic modifications in cultured hippocampal neurons: dependence on spike timing, synaptic strength, and postsynaptic cell type,” J. Neurosci., 18, 10464–10472 (1998).PubMedGoogle Scholar
  26. 26.
    D. L. Adkins, J. Boychuk, M. S. Remple, and J. A. Kleim, “Motor training induces experience-specific patterns of plasticity across motor cortex and spinal cord,” J. Appl. Physiol., 101, 1776–1782 (2006).PubMedCrossRefGoogle Scholar
  27. 27.
    D. A. Gusnard, E. Akbudak, G. L. Shulman, and M. E. Raichle, “Medial prefrontal cortex and self-referential mental activity: relation to a default mode of brain function,” Proc. Natl. Acad. Sci. USA, 98, 4259–4264 (2001).PubMedCrossRefGoogle Scholar
  28. 28.
    C. Herry and N. Mons, “Resistance to extinction is associated with impaired immediate early gene induction in medial prefrontal cortex and amygdale,” Eur. J. Neurosci., 20, 781–790 (2004).PubMedCrossRefGoogle Scholar
  29. 29.
    M. S. Rioult-Pedotti, D. Friedman, G. Hess, and J. P. Donoghue, “Strengthening of horizontal cortical connections following skill learning,” Nat. Neurosci., 1, 230–234 (1998).PubMedCrossRefGoogle Scholar
  30. 30.
    G. S. Withers and W. T. Greenough, “Reach training selectively alters dendritic branching in subpopulations of layer II-III pyramids in rat motor-somatosensory forelimb cortex,” Neuropsychologia, 27, 61–69 (1989).PubMedCrossRefGoogle Scholar
  31. 31.
    C. Pavlides, E. Miyashita, and H. Asanuma, “Projection from the sensory to the motor cortex is important in learning motor skills in the monkey,” J. Neurophysiol., 70, 733–741 (1993).PubMedGoogle Scholar
  32. 32.
    L. L. Porter and K. Sakamoto, “Organization and synaptic relationships of the projection from the primary sensory to the primary motor cortex in the cat,” J. Comp. Neurol., 271, 387–396 (1988).PubMedCrossRefGoogle Scholar
  33. 33.
    H. Wenk, “The nucleus basalis magnocellularis Meynert (NbmM) complex – a central integrator of coded ‘limbic signals’ linked to neocortical modular operation? A proposed (heuristic) model of function,” J. Hirnforsch., 30, 127–151 (1989).PubMedGoogle Scholar
  34. 34.
    O. V. Vlasenko, V. A. Maisky, A. V. Maznychenko, and A. I. Pilyavskii, “NADPH-diaphorase activity and neurovascular coupling in the rat cerebral cortex,” Fiziol. Zh., 54, No. 1, 45–53 (2008).PubMedGoogle Scholar
  35. 35.
    J. M. Moscarello, O. Ben-Shahar, and A. Ettenberg, “Dynamic interaction between medial prefrontal cortex and nucleus accumbens as a function of both motivational state and reinforced magnitude: a c-Fos immunocytochemistry study,” Brain Res., 1169, 69–76 (2007).PubMedCrossRefGoogle Scholar
  36. 36.
    A. K. Majewska and M. Sur, “Plasticity and specificity of cortical processing networks,” Trends Neurosci., 29, 323–329 (2006).PubMedCrossRefGoogle Scholar
  37. 37.
    A. R. Luft, M. M. Buitrago, T. Ringer, et al., “Motor skill learning depends on protein synthesis in motor cortex after training,” J. Neurosci., 24, 6515–6520 (2004).PubMedCrossRefGoogle Scholar
  38. 38.
    H. Girouard and C. Iadecola, “Neurovascular coupling in the normal brain and in hypertension, stroke, and Alzheimer disease,” J. Appl. Physiol., 100, 328–335 (2006).PubMedCrossRefGoogle Scholar
  39. 39.
    B. Cauli, X.-K. Tong, A. Rancillac, et al., “Cortical GABA interneurons in neurovascular coupling: relays for subcortical vasoactive pathways,” J. Neurosci., 24, 8940–8949 (2004).PubMedCrossRefGoogle Scholar
  40. 40.
    X.-K. Tong and E. Hamel, “Regional cholinergic denervation of cortical microvessels and nitric oxide synthase-containing neurons in Alzheimer’s disease,” Neuroscience, 92, 163–175 (1999).PubMedCrossRefGoogle Scholar
  41. 41.
    R. A. Swain, A. B. Harris, E. C. Wiener, et al., “Prolonged exercise induces angiogenesis and increases cerebral blood volume in primary motor cortex of the rat,” Neuroscience, 117, 1037–1046 (2003).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2008

Authors and Affiliations

  • O. V. Vlasenko
    • 1
  • A. I. Pilyavskii
    • 2
  • V. A. Maiskii
    • 2
  • A. V. Maznichenko
    • 2
  1. 1.Pirogov Vinnitsa National Medical University, Ministry of Public HealthVinnitsaUkraine
  2. 2.Bogomolets Institute of PhysiologyNational Academy of Sciences of UkraineKyivUkraine

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