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@ARTICLE{Cremer:7022,
      author       = {Cremer, C.M. and Cremer, M. and Lopez Escobar, J. and
                      Speckmann, E.J. and Zilles, K.},
      title        = {{F}ast, quantitative in situ hybridization of rare m{RNA}s
                      using 14{C}-standards and phosphorus imaging},
      journal      = {Journal of neuroscience methods},
      volume       = {185},
      issn         = {0165-0270},
      address      = {Amsterdam [u.a.]},
      publisher    = {Elsevier Science},
      reportid     = {PreJuSER-7022},
      pages        = {56 - 61},
      year         = {2009},
      note         = {Record converted from VDB: 12.11.2012},
      abstract     = {The use of radiolabelled probes for in situ hybridization
                      (ISH) bears the advantage of high sensitivity and
                      quantifiability. The crucial disadvantages are laborious
                      hybridization protocols, exposition of hybridized sections
                      to film for up to several weeks and the time consuming need
                      to prepare tissue standards with relatively short-lived
                      isotopes like (33)P or (35)S for each experiment. The
                      quantification of rare mRNAs like those encoding for
                      subunits of neurotransmitter receptors is therefore a
                      challenge in ISH. Here, we describe a method for fast,
                      quantitative in situ hybridization (qISH) of mRNAs using
                      (33)P-labelled oligonucleotides together with (14)C-polymer
                      standards (Microscales, Amersham Biosciences) and a
                      phosphorus imaging system (BAS 5000 BioImage Analyzer,
                      Raytest-Fuji). It enables a complete analysis of rare mRNAs
                      by ISH. The preparation of short-lived (33)P-standards for
                      each experiment was replaced by co-exposition and
                      calibration of long-lived (14)C-standards together with
                      (33)P-labelled brain paste standards. The use of a
                      phosphorus imaging system allowed a reduction of exposition
                      time following hybridization from several weeks to a few
                      hours or days. We used this approach as an example for
                      applications to quantify the expression of GluR1 and GluR2
                      subunit mRNAs of the
                      alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
                      (AMPA) receptor in the hippocampus of untreated rats, and
                      after intraperitoneal application of the organo-arsenic
                      compound dimethyl arsenic acid.},
      keywords     = {Animals / Cacodylic Acid: toxicity / Carbon Radioisotopes:
                      chemistry / Hippocampus: drug effects / Hippocampus:
                      metabolism / Hippocampus: physiopathology / Image
                      Processing, Computer-Assisted: instrumentation / Image
                      Processing, Computer-Assisted: methods / In Situ
                      Hybridization: methods / Male / Neurochemistry: methods /
                      Oligonucleotides: chemistry / Phosphorus Radioisotopes:
                      chemistry / RNA, Messenger: analysis / RNA, Messenger:
                      metabolism / Rats / Rats, Wistar / Receptors, AMPA: genetics
                      / Reference Standards / Time Factors / Carbon Radioisotopes
                      (NLM Chemicals) / Oligonucleotides (NLM Chemicals) /
                      Phosphorus Radioisotopes (NLM Chemicals) / RNA, Messenger
                      (NLM Chemicals) / Receptors, AMPA (NLM Chemicals) /
                      glutamate receptor ionotropic, AMPA 1 (NLM Chemicals) /
                      glutamate receptor ionotropic, AMPA 2 (NLM Chemicals) /
                      Cacodylic Acid (NLM Chemicals) / J (WoSType)},
      cin          = {INM-2 / JARA-BRAIN},
      ddc          = {610},
      cid          = {I:(DE-Juel1)INM-2-20090406 / $I:(DE-82)080010_20140620$},
      pnm          = {Funktion und Dysfunktion des Nervensystems},
      pid          = {G:(DE-Juel1)FUEK409},
      shelfmark    = {Biochemical Research Methods / Neurosciences},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:19761793},
      UT           = {WOS:000272473000008},
      doi          = {10.1016/j.jneumeth.2009.09.010},
      url          = {https://juser.fz-juelich.de/record/7022},
}