Skip to main content
SpringerLink
Log in

Measurement by ELISA of Complement Factor 4 (C4) in the Rat Brain: Necessity for Removal of Cerebrovascular Proteins

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Assessment of complement 4 (C4) levels in experimental animals is used as a marker for activation of the classical complement pathway. The objective of this study was to develop a method for measuring C4 concentrations in the rat brain. An ELISA (sensitivity = 0.5 ng C4/ml) was used to measure C4 in regional brain homogenates from Fischer rats cardiac-perfused with phosphate buffered saline to remove cerebrovascular contents, and from sham-perfused rats. Ventral midbrain C4 levels were increased (p < 0.001) versus frontal cortex and striatum in sham-perfused rats, whereas after perfusion there were no differences between brain regions. Removal of cerebrovascular contents decreased C4 by 43% in striatum, 52% in frontal cortex, and 69% in ventral midbrain (all p < 0.01 versus sham-perfused means). These results indicate that C4 in the rat brain can be measured quantitatively by ELISA provided that cerebrovascular proteins are removed by perfusion.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Morgan BP, Harris CL (2003) Complement therapeutics; history and current progress. Mol Immunol 40:159–170

    Article  PubMed  CAS  Google Scholar 

  2. Kirschfink M (2001) Targeting complement in therapy. Immunol Rev 180:177–189

    Article  PubMed  CAS  Google Scholar 

  3. D’Ambrosio AL, Pinsky DJ, Connolly ES (2001) The role of the complement cascade in ischemia/reperfusion injury: implications for neuroprotection. Mol Med 7:367–382

    PubMed  CAS  Google Scholar 

  4. McGeer PL, Akiyama H, Itagaki S, (1989) Activation of the classical complement pathway in brain tissue of Alzheimer patients. Neurosci Lett 107:341–346

    Article  PubMed  CAS  Google Scholar 

  5. Yamada T, McGeer PL, McGeer EG (1992) Lewy bodies in Parkinson’s disease are recognized by antibodies to complement proteins. Acta Neuropathol (Berl) 84:100–104

    Article  CAS  Google Scholar 

  6. Yamada T, Moroo I, Koguchi Y et al (1994) Increased concentration of C4d complement protein in the cerebrospinal fluids in progressive supranuclear palsy. Acta Neurol Scand 89:42–46

    Article  PubMed  CAS  Google Scholar 

  7. Matsuoka Y, Picciano M, Malester B et al (2001) Inflammatory responses to amyloidosis in a transgenic mouse model of Alzheimer’s disease. Am J Pathol 158:1345–1354

    PubMed  CAS  Google Scholar 

  8. Sinigaglia-Coimbra R, Cavalheiro EA, Coimbra CG (2002) Postischemic hyperthermia induces Alzheimer-like pathology in the rat brain. Acta Neuropathol (Berl) 103:444–452

    Article  CAS  Google Scholar 

  9. Fonseca MI, Zhou J, Botto M et al (2004) Absence of C1q leads to less neuropathology in transgenic mouse models of Alzheimer’s disease. J Neurosci 24:6457–6465

    Article  PubMed  CAS  Google Scholar 

  10. Spielman L, Winger D, Ho L et al (2002) Induction of the complement component C1qB in brain of transgenic mice with neuronal overexpression of human cyclooxygenase-2. Acta Neuropathol (Berl) 103:157–162

    Article  CAS  Google Scholar 

  11. Cowell RM, Plane JM, Silverstein FS (2003) Complement activation contributes to hypoxic–ischemic brain injury in neonatal rats. J Neurosci 23:9459–9468

    PubMed  Google Scholar 

  12. Loeffler DA, Linder MC, Zamany M et al (1999) Measurement of acute phase proteins in the rat brain: contribution of vascular contents. Neurochem Res 24:1313–1317

    Article  PubMed  CAS  Google Scholar 

  13. Bos IG, van Mierlo GJ, Bleeker WK et al (2001) The potentiation of human C1-inhibitor by dextran sulphate is transient in vivo: studies in a rat model. Int Immunopharmacol 1:1583–1595

    Article  PubMed  CAS  Google Scholar 

  14. Roos A, Nauta AJ, Broers D et al (2001) Specific inhibition of the classical complement pathway by C1q-binding peptides. J Immunol 167:7052–7059

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Thanks are expressed to Donna Selenich, Elizabeth Debeliso, Diane Farrah, and Stephanie Conant, Ph.D., for technical assistance. This investigation was funded by a contribution from Mrs. Martha Loeffler in memory of Drs. Erwin and Harold Loeffler.

Author information

Authors and Affiliations

  1. Division of Neurology, William Beaumont Hospital Research Institute, 3811 West Thirteen Mile Rd., Royal Oak, MI, 48073, USA

    David A. Loeffler & Dianne M. Camp

Authors
  1. David A. Loeffler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dianne M. Camp
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to David A. Loeffler.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Loeffler, D.A., Camp, D.M. Measurement by ELISA of Complement Factor 4 (C4) in the Rat Brain: Necessity for Removal of Cerebrovascular Proteins . Neurochem Res 31, 999–1002 (2006). https://doi.org/10.1007/s11064-006-9105-z

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-006-9105-z

Keywords

Search

Navigation