Intracranial landmarks and other techniques to further improve the precision of stereotaxic tracer injections


This study is meant to combine traditional aspects of tracer microinjections using bone landmarks like bregma and lambda with novel procedures in which specific parts of the brain can serve as reference points. For telencephalic and diencephalic injections, the brain surface, the interhemispheric groove and the straight sinus can be used as absolute zero points for dorso-ventral, medio-lateral and rostro-caudal coordinates, respectively. In case of brainstem targets, the surface of the rhomboid fossa, the posterior spinal artery and the obex could serve as reference points along the above-mentioned coordinates. The application of high-precision stereotaxic measurements based on intracranial landmarks and sophisticated surgical procedures can yield well-targeted, small and well circumscribed injection sites that make possible the mapping of discrete nuclear subdivisions or delicate nuclei in the brain.

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  1. Boldogköi ZS, Reichart A, Tóth EI, Sik A, Erdélyi F, Medveczky I, Llorens-Cortes C, Palkovits M, Lenkei ZS (2002) Construction of recombinant pseudorabies viruses optimized for labeling and neurochemical characterization of neural circuitry. Mol Brain Res 109:105–118

  2. Boldogköi ZS, Sik A, Dénes Á, Reichart A, Toldi J, Gerendai I, Kovács JK, Palkovits M (2004) Novel tracing paradigms-genetically engineered herpesviruses as tools for mapping functional circuits within the CNS: present status and future prospects. Prog Neurobiol 72:417–445

  3. Gerfen CR, Sawchenko PE (1984) An anterograde neuroanatomical tracing method that shows the detailed morphology of neurons, their axons and terminals: immunohistochemical localization of an axonally transported plant lectin, Phaseolus Vulgaris Leucoagglutinin (PHA-L). Brain Res 290:219–238

  4. Gildenberg LP (1988) Stereotactic surgery: present and past stereotactic neurosurgery. Williams and Wilkins, Baltimore

  5. Kelly JP (1991) Introduction and historical aspects tumor stereotaxis. W.B. Saunders Company, Philadelphia

  6. Kovács ZI, Palkovits M (2010) Microcapillary specifically designed for pressure microinjections of very low volumes. J Neurosci Methods 190(2):229–234

  7. Levy R (1992) A short history of stereotactic surgery. Cyber Mus Neurosurg. Accessed 01 March 2010

  8. Luppi PH, Fort P, Jouvet M (1990) Iontophoretic application of unconjugated cholera toxin B subunit (CTb) combined with immunohistochemistry of neurochemical substances: a method for transmitter identification of retrogradely labeled neurons. Brain Res 534:209–224

  9. Palkovits M, Kiss JZ, Beinfeld MC, Williams TH (1982) Cholecystokinin in the nucleus of the solitary tract of the rat. Brain Res 252:386–390

  10. Palkovits M, Mezey É, Eskay RL, Brownstein MJ (1986) Innervation of the nucleus of the solitary tract and the dorsal vagal nucleus by thyrotropin-releasing hormone-containing raphe neurons. Brain Res 373:246–251

  11. Paxinos G, Franklin KBJ (1997) The mouse brain in stereotaxic coordinates. Academic Press, Sydney

  12. Paxinos G, Watson C (2005) The rat brain in stereotaxic coordinates, 5th edn. Academic Press, Sydney

  13. Ritter S, Dinh TT, Zhang Y (2000) Localization of hindbrain glucoreceptive sites controlling food intake and blood glucose. Brain Res 856(1–2):37–47

  14. Swanson LW (2004). Brain maps: structure of the rat brain, 3rd edn. Elsevier, Amsterdam

  15. Veenman CL, Reiner A, Honig MG (1992) Biotinylated dextran amine as an anterograde tracer for single- and double-labeling studies. J Neurosci Methods 41:239–254

  16. Xiao J (2007) A new coordinate system for rodent brain and variability in the brain weights and dimensions of different ages in the naked mole-rat. J Neurosci Methods 162:162–170

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This research was supported by grants from the Hungarian Science Research Foundation, OTKA TS-049861. Special thanks to, Zsuzsanna E. Tóth PhD, Zoltán Buzogány D. D. and János Cifra M. D. for providing us substances (ZnCl2) and advice.

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Correspondence to Zsolt I. Kovács.

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Kovács, Z.I., Palkovits, M. Intracranial landmarks and other techniques to further improve the precision of stereotaxic tracer injections. Exp Brain Res 208, 51–60 (2011) doi:10.1007/s00221-010-2459-0

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  • Cranial landmarks
  • Intracranial landmarks
  • Straight sinus
  • Obex
  • Virus microinjections
  • Brain mapping
  • Interhemispheric groove/fissure