Skip to main content
PMC home page
Neuroscience Bulletin logoLink to Neuroscience Bulletin
. 2015 Sep 7;31(6):685–696. doi: 10.1007/s12264-015-1558-0

Increasing the effectiveness of intracerebral injections in adult and neonatal mice: a neurosurgical point of view

Bertrand Mathon 1,2,3,4,5,✉, Mérie Nassar 2,3,4,5, Jean Simonnet 2,3,4,5, Caroline Le Duigou 2,3,4,5, Stéphane Clemenceau 1, Richard Miles 2,3,4,5, Desdemona Fricker 2,3,4,5
PMCID: PMC5563725  PMID: 26345180

Abstract

Intracerebral injections of tracers or viral constructs in rodents are now commonly used in the neurosciences and must be executed perfectly. The purpose of this article is to update existing protocols for intracerebral injections in adult and neonatal mice. Our procedure for stereotaxic injections in adult mice allows the investigator to improve the effectiveness and safety, and save time. Furthermore, for the first time, we describe a two-handed procedure for intracerebral injections in neonatal mice that can be performed by a single operator in a very short time. Our technique using the stereotaxic arm allows a higher precision than freehand techniques previously described. Stereotaxic injections in adult mice can be performed in 20 min and have >90% efficacy in targeting the injection site. Injections in neonatal mice can be performed in 5 min. Efficacy depends on the difficulty of precisely localizing the injection sites, due to the small size of the animal. We describe an innovative, effortless, and reproducible surgical protocol for intracerebral injections in adult and neonatal mice.

Keywords: rodents, surgery, neurosciences, stereotaxic injection, newborn mice, neonatal brain

References

  • [1].Cardin JA, Carlen M, Meletis K, Knoblich U, Zhang F, Deisseroth K, et al. Targeted optogenetic stimulation and recording of neurons in vivo using cell-type-specific expression of Channelrhodopsin-2. Nat Protoc. 2010;5:247–254. doi: 10.1038/nprot.2009.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [2].Puntel M, Kroeger KM, Sanderson NS, Thomas CE, Castro MG, Lowenstein PR. Curr Protoc Neurosci. 2010. Gene transfer into rat brain using adenoviral vectors. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [3].Cetin A, Komai S, Eliava M, Seeburg PH, Osten P. Stereotaxic gene delivery in the rodent brain. Nat Protoc. 2006;1:3166–3173. doi: 10.1038/nprot.2006.450. [DOI] [PubMed] [Google Scholar]
  • [4].Davidson S, Truong H, Nakagawa Y, Giesler G Jr. A microinjection technique for targeting regions of embryonic and neonatal mouse brain in vivo. Brain Res. 2010;1307:43–52. doi: 10.1016/j.brainres.2009.10.024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [5].Li J, Daly TM. Adeno-associated virus-mediated gene transfer to the neonatal brain. Methods. 2002;28:203–207. doi: 10.1016/S1046-2023(02)00224-4. [DOI] [PubMed] [Google Scholar]
  • [6].Pilpel N, Landeck N, Klugmann M, Seeburg PH, Schwarz MK. Rapid, reproducible transduction of select forebrain regions by targeted recombinant virus injection into the neonatal mouse brain. J Neurosci Methods. 2009;182:55–63. doi: 10.1016/j.jneumeth.2009.05.020. [DOI] [PubMed] [Google Scholar]
  • [7].Shurey S, Akelina Y, Legagneux J, Malzone G, Jiga L, Ghanem AM. The rat model in microsurgery education: classical exercises and new horizons. Arch Plast Surg. 2014;41:201–208. doi: 10.5999/aps.2014.41.3.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [8].Albrecht M, Henke J, Tacke S, Markert M, Guth B. Effects of isoflurane, ketamine-xylazine and a combination of medetomidine, midazolam and fentanyl on physiological variables continuously measured by telemetry in Wistar rats. BMC Vet Res. 2014;10:198. doi: 10.1186/s12917-014-0198-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [9].Redfors B, Shao Y, Omerovic E. Influence of anesthetic agent, depth of anesthesia and body temperature on cardiovascular functional parameters in the rat. Lab Anim. 2014;48:6–14. doi: 10.1177/0023677213502015. [DOI] [PubMed] [Google Scholar]
  • [10].Hartsfield SM. Advantages and guidelines for using ketamine for induction of anesthesia. Vet Clin North Am Small Anim Pract. 1992;22:266–267. doi: 10.1016/S0195-5616(92)50604-0. [DOI] [PubMed] [Google Scholar]
  • [11].McDowell A, Fothergill JA, Khan A, Medlicott NJ. A cyclodextrin formulation to improve use of the anesthetic tribromoethanol (Avertin((R))) J Pharm Bioallied Sci. 2014;6:16–21. doi: 10.4103/0975-7406.124303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [12].Meyer RE, Fish RE. A review of tribromoethanol anesthesia for production of genetically engineered mice and rats. Lab Anim (NY) 2005;34:47–52. doi: 10.1038/laban1105-47. [DOI] [PubMed] [Google Scholar]
  • [13].Haberl MG, Viana da Silva S, Guest JM, Ginger M, Ghanem A, Mulle C, et al. Brain Struct Funct. 2014. An anterograde rabies virus vector for high-resolution large-scale reconstruction of 3D neuron morphology. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [14].Atasoy D, Aponte Y, Su HH, Sternson SM. A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long–range circuit mapping. J Neurosci. 2008;28:7025–7030. doi: 10.1523/JNEUROSCI.1954-08.2008. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Neuroscience Bulletin are provided here courtesy of Springer

RESOURCES