Child's Nervous System

, Volume 23, Issue 10, pp 1171–1179 | Cite as

Diffuse alterations in synaptic protein expression following focal traumatic brain injury in the immature rat

  • G. T. Gobbel
  • C. Bonfield
  • E. B. Carson-Walter
  • P. D. Adelson
Original Paper



The mechanisms responsible for cognitive decline after traumatic brain injury (TBI) in pediatric patients are poorly understood. The present study examined the potential role of synaptic alterations in this process by using an animal model of immature head injury to define the impact of TBI on expression of the synaptic protein, synaptophysin.

Materials and methods

After craniotomy, TBI was induced in postnatal day 17 (PND17) rats using controlled cortical impact delivered to the left hemisphere. NeuN, a neuronal marker, and synaptophysin expression were examined 1 day, 1 week, and 1 month after injury by immunohistochemistry and immunoblotting.


There were significant decreases in both NeuN and synaptophysin after 1 day and 1 week but not 1 month after injury within the hippocampus and neocortex adjacent to the impact site compared to sham-injured controls. The decrease in synaptophysin and NeuN was also noted in the contralateral hippocampus by 1 day after injury and in the contralateral neocortex by 1 week, indicating that changes in protein expression were not solely localized to the injury site but occurred in more distant regions as well.


In conclusion, the decrease and recovery in synaptophysin parallel the cognitive changes that occur after experimental TBI in the PND17 rat, which suggests that changes in this protein may contribute to cognitive declines after injury. The results also suggest that, in spite of the focal nature of the impact, diffuse alterations in protein expression can occur after immature TBI and may contribute to the subsequent cognitive dysfunction.


Pediatric Rat NeuN Synaptophysin Traumatic brain injury 


  1. 1.
    Adelson PD, Dixon CE, Davis DS, Rodriguez, AG, Tran MP, Jenkins LW, Kochanek PM (2001) Differential age-at-injury effect of NMDA blockade on outcome following controlled cortical impact in immature rats. J Neurotrauma 18:1143CrossRefGoogle Scholar
  2. 2.
    Andrews RJ (1991) Transhemispheric diaschisis. A review and comment. Stroke 22:943–949PubMedGoogle Scholar
  3. 3.
    Bayer SA (1980) Development of the hippocampal region in the rat. II. Morphogenesis during embryonic and early postnatal life. J Comp Neurol 190:115–134PubMedCrossRefGoogle Scholar
  4. 4.
    Bayer, SA, Altman J, Russo RJ, Zhang X (1993) Timetables of neurogenesis in the human brain based on experimentally determined patterns in the rat. Neurotoxicology 14:83–144PubMedGoogle Scholar
  5. 5.
    Beers SR (1992) Cognitive effects of mild head injury in children and adolescents. Neuropsychol Rev 3:281–320PubMedCrossRefGoogle Scholar
  6. 6.
    Bourgeois JP, Goldman-Rakic PS, Rakic P (1994) Synaptogenesis in the prefrontal cortex of rhesus monkeys. Cereb Cortex 4:78–96PubMedCrossRefGoogle Scholar
  7. 7.
    Bruce DA, Schut L, Bruno LA, Wood JH, Sutton LN (1978) Outcome following severe head injuries in children. J Neurosurg 48:679–688PubMedGoogle Scholar
  8. 8.
    Buckley K, Kelly RB (1985) Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells. J Cell Biol 100:1284–1294PubMedCrossRefGoogle Scholar
  9. 9.
    Calhoun ME, Jucker M, Martin LJ, Thinakaran G, Price DL, Mouton PR (1996) Comparative evaluation of synaptophysin-based methods for quantification of synapses. J Neurocytol 25:821–828PubMedCrossRefGoogle Scholar
  10. 10.
    Calhoun ME, Kurth D, Phinney AL, Long JM, Hengemihle J, Mouton PR, Ingram DK, Jucker M (1998) Hippocampal neuron and synaptophysin-positive bouton number in aging C57BL/6 mice. Neurobiol Aging 19:599–606PubMedCrossRefGoogle Scholar
  11. 11.
    Chen KS, Masliah E, Mallory M, Gage FH (1995) Synaptic loss in cognitively impaired aged rats is ameliorated by chronic human nerve growth factor infusion. Neuroscience 68:19–27PubMedCrossRefGoogle Scholar
  12. 12.
    Deweer B, Pillon B, Pochon JB, Dubois B (2001) Is the HM story only a ‘remote memory’? Some facts about hippocampus and memory in humans. Behav Brain Res 127:209–224PubMedCrossRefGoogle Scholar
  13. 13.
    Fletcher JM, Ewing-Cobbs L, Francis DJ, Levin HS (1995) Variability in outcomes after traumatic brain injury in children: a developmental perspective. In: Broman SH, Michel ME (eds) Traumatic head injury in children. Oxford University Press, New York, pp 3–21Google Scholar
  14. 14.
    Hagberg H, Bona E, Gilland E, Puka-Sundvall M (1997) Hypoxia-ischaemia model in the 7-day-old rat: possibilities and shortcomings. Acta Paediatr Suppl 422:85–88PubMedGoogle Scholar
  15. 15.
    Hunter SF, Leavitt JA, Rodriguez M (1997) Direct observation of myelination in vivo in the mature human central nervous system. A model for the behaviour of oligodendrocyte progenitors and their progeny. Brain 120(Pt 11):2071–2082PubMedCrossRefGoogle Scholar
  16. 16.
    Ikonomidou C, Bosch F, Miksa M, Bittigau P, Vockler J, Dikranian K, Tenkova TI, Stefovska V, Turski L, Olney JW (1999) Blockade of NMDA receptors and apoptotic neurodegeneration in the developing brain. Science 283:70–74PubMedCrossRefGoogle Scholar
  17. 17.
    Iwasaki N, Hamano K, Okada Y, Horigome Y, Nakayama J, Takeya T, Takita H, Nose T (1997) Volumetric quantification of brain development using MRI. Neuroradiology 39:841–846PubMedCrossRefGoogle Scholar
  18. 18.
    Jacobson M (1991) Formation of dendrites and development of synaptic connections. In: Jacobson M (ed) Developmental neurobiology. Plenum, New York, pp 223–284Google Scholar
  19. 19.
    Jahn R, Schiebler W, Ouimet C, Greengard P (1985) A 38,000-dalton membrane protein (p38) present in synaptic vesicles. Proc Natl Acad Sci USA 82:4137–4141PubMedCrossRefGoogle Scholar
  20. 20.
    Kraus JF, Fife D, Conroy C (1987) Pediatric brain injuries: the nature, clinical course, and early outcomes in a defined United States’ population. Pediatrics 79:501–507PubMedGoogle Scholar
  21. 21.
    Levin HS (1985) General considerations and neurobehavioral recovery. Part II. Neurobehavioral recovery. In: Becker DP, Povlishock JT (eds) Central nervous suystem status report. NINCDS and NIH, Washington, D.C., pp 281–299Google Scholar
  22. 22.
    Levin HS, Ewing-Cobbs, Eisenberg HM (1995) Neurobehavioral outcome of pediatric closed head injury. In: Broman SH, Michel ME (eds) Traumatic head injury in children. Oxford University Press, New York, pp 70–94Google Scholar
  23. 23.
    Levin HS, Aldrich EF, Saydjari C, Eisenberg HM, Foulkes MA, Bellefleur M, Luerssen TG, Jane JA, Marmarou A, Marshall LF, Al E (1992) Severe head injury in children: experience of the traumatic coma data bank. Neurosurgery 31:435–443, discussion 443–434PubMedCrossRefGoogle Scholar
  24. 24.
    Lind D, Franken S, Kappler J, Jankowski J, Schilling K (2005) Characterization of the neuronal marker NeuN as a multiply phosphorylated antigen with discrete subcellular localization. J Neurosci Res 79:295–302PubMedCrossRefGoogle Scholar
  25. 25.
    Liu HX, Zhang JJ, Zheng P, Zhang Y (2005) Altered expression of MAP-2, GAP-43, and synaptophysin in the hippocampus of rats with chronic cerebral hypoperfusion correlates with cognitive impairment. Brain Res Mol Brain Res 139:169–177PubMedCrossRefGoogle Scholar
  26. 26.
    Lohmann SM, Ueda T, Greengard P (1978) Ontogeny of synaptic phosphoproteins in brain. Proc Natl Acad Sci USA 75:4037–4041PubMedCrossRefGoogle Scholar
  27. 27.
    Luerssen TG, Klauber MR, Marshall LF (1988) Outcome from head injury related to patient’s age. A longitudinal prospective study of adult and pediatric head injury. J Neurosurg 68:409–416PubMedCrossRefGoogle Scholar
  28. 28.
    Masliah E, Fagan AM, Terry RD, Deteresa R, Mallory M, Gage FH (1991) Reactive synaptogenesis assessed by synaptophysin immunoreactivity is associated with GAP-43 in the dentate gyrus of the adult rat. Exp Neurol 113:131–142PubMedCrossRefGoogle Scholar
  29. 29.
    Mcphail LT, McBride CB, Mcgraw J, Steeves JD, Tetzlaff W (2004) Axotomy abolishes NeuN expression in facial but not rubrospinal neurons. Exp Neurol 185:182–190PubMedCrossRefGoogle Scholar
  30. 30.
    Pohl D, Bittigau P, Ishimaru MJ, Stadthaus D, Hubner C, Olney JW, Turski L, Ikonomidou C (1999) N-Methyl-d-aspartate antagonists and apoptotic cell death triggered by head trauma in developing rat brain. Proc Natl Acad Sci USA 96:2508–2513PubMedCrossRefGoogle Scholar
  31. 31.
    Rakic P, Bourgeois JP, Eckenhoff MF, Zecevic N, Goldman-Rakic PS (1986) Concurrent overproduction of synapses in diverse regions of the primate cerebral cortex. Science 232:232–235PubMedCrossRefGoogle Scholar
  32. 32.
    Rice D, Barone S Jr (2000) Critical periods of vulnerability for the developing nervous system: evidence from humans and animal models. Environ Health Perspect 108(Suppl 3):511–533PubMedCrossRefGoogle Scholar
  33. 33.
    Shojo H, Kibayashi K (2006) Changes in localization of synaptophysin following fluid percussion injury in the rat brain. Brain Res 1078:198–211PubMedCrossRefGoogle Scholar
  34. 34.
    Smith ML (1989) Memory disorders associated with temporal lobe lesions. In: Boller F, Grafman J (eds) Handbook of neuropsychology. Elsevier, pp 91–106Google Scholar
  35. 35.
    Smith TD, Adams MM, Gallagher M, Morrison JH, Rapp PR (2000) Circuit-specific alterations in hippocampal synaptophysin immunoreactivity predict spatial learning impairment in aged rats. J Neurosci 20:6587–6593PubMedGoogle Scholar
  36. 36.
    Stevenson, KL, Skinner JC, Davis DS, Tran MP, Dixon CE, Kochanek PM, Jenkins LW, Adelson PD (2000) Behavioral dysfunction in immature rats after controlled cortical impact (CCI). J Neurotrauma 17:944Google Scholar
  37. 37.
    Tran MP, Rodriguez AG, Dixon CE, Kochanek PM, Davis DS, Stevenson KL, Jenkins LW, Adelson PD (2001) Histologic effects of acute NMDA blockade following controlled cortical impact in immature rats. J Neurotrauma 18:1141Google Scholar
  38. 38.
    Unal-Cevik I, Kilinc M, Gursoy-Ozdemir Y, Gurer G, Dalkara T (2004) Loss of NeuN immunoreactivity after cerebral ischemia does not indicate neuronal cell loss: a cautionary note. Brain Res 1015:169–174PubMedCrossRefGoogle Scholar
  39. 39.
    Uylings HB, Van Eden CG (1990) Qualitative and quantitative comparison of the prefrontal cortex in rat and in primates, including humans. Prog Brain Res 85:31–62PubMedCrossRefGoogle Scholar
  40. 40.
    Voigt T, De Lima AD, Beckmann M (1993) Synaptophysin immunohistochemistry reveals inside-out pattern of early synaptogenesis in ferret cerebral cortex. J Comp Neurol 330:48–64PubMedCrossRefGoogle Scholar
  41. 41.
    Wiedenmann B, Franke WW (1985) Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynaptic vesicles. Cell 41:1017–1028PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • G. T. Gobbel
    • 1
  • C. Bonfield
    • 1
    • 2
  • E. B. Carson-Walter
    • 1
  • P. D. Adelson
    • 1
  1. 1.Department of Neurological SurgeryUniversity of PittsburghPittsburghUSA
  2. 2.PittsburghUSA

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