Polyclonal antibodies to LIM proteins CRP2 and CRIP2 reveal their subcellular localizations in olfactory precursor cells

Biochemistry (Mosc). 2009 Mar;74(3):336-41. doi: 10.1134/s0006297909030134.

Abstract

In this study, we describe the presence of CRP2 (cysteine- and glycine-rich protein 2) and CRIP2 (cysteine-rich intestinal protein 2), which are members of group 2 LIM proteins, in rat olfactory precursor cells by reverse transcription polymerase chain reaction. We have developed polyclonal antibodies against CRP2 and CRIP2 individually. Specificity of the antibodies was demonstrated by Western blot analysis, using CRP2 and CRIP2 transfected cells. No cross-reactivity was observed between the antibodies. Furthermore, we used the antibodies to determine the expression and localization of CRP2 and CRIP2 in olfactory precursor cells by Western blot analysis and immunofluorescence staining. Our results demonstrated that in undifferentiated olfactory precursor cells CRP2 was distributed both in the nucleus and the cytoplasm, whereas CRIP2 was predominantly localized in the cytoplasm. While the olfactory precursor cells differentiated into end cells, only the expression of CRIP2 would be detected. The function of these LIM proteins in olfactory precursor cells warrants further study.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / analysis*
  • Antibodies, Monoclonal / immunology
  • Antibody Specificity / immunology
  • Base Sequence
  • Blotting, Western
  • COS Cells
  • Carrier Proteins / genetics
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism*
  • Cell Nucleus / metabolism
  • Cells, Cultured
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cytoplasm / metabolism
  • Fluorescent Antibody Technique
  • LIM Domain Proteins
  • Muscle Proteins / genetics
  • Muscle Proteins / immunology
  • Muscle Proteins / metabolism*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / immunology
  • Nuclear Proteins / metabolism*
  • Olfactory Bulb / cytology
  • Olfactory Bulb / metabolism*
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection

Substances

  • Antibodies, Monoclonal
  • Carrier Proteins
  • Crip2 protein, rat
  • Csrp2 protein, rat
  • LIM Domain Proteins
  • Muscle Proteins
  • Nuclear Proteins