Home LiteratureArticle Details
PMID: 15208716 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Functional fluorescent Ca2+ indicator proteins in transgenic mice under TET control.

PLoS biology ·Vol. 2 ·No. 6 ·2004-06-00 ·Pages e163

Hasan MT, Friedrich RW, Euler T, Larkum ME, Giese G, Both M, Duebel J, Waters J, Bujard H, Griesbeck O, Tsien RY, Nagai T, Miyawaki A, Denk W

Abstract

Genetically encoded fluorescent calcium indicator proteins (FCIPs) are promising tools to study calcium dynamics in many activity-dependent molecular and cellular processes. Great hopes-for the measurement of population activity, in particular-have therefore been placed on calcium indicators derived from the green fluorescent protein and their expression in (selected) neuronal populations. Calcium transients can rise within milliseconds, making them suitable as reporters of fast neuronal activity. We here report the production of stable transgenic mouse lines with two different functional calcium indicators, inverse pericam and camgaroo-2, under the control of the tetracycline-inducible promoter. Using a variety of in vitro and in vivo assays, we find that stimuli known to increase intracellular calcium concentration (somatically triggered action potentials (APs) and synaptic and sensory stimulation) can cause substantial and rapid changes in FCIP fluorescence of inverse pericam and camgaroo-2.

MeSH Terms
Animals Brain/metabolism Brain Mapping/methods Calcium/metabolism,physiology Calcium Channels/metabolism,physiology Doxycycline Fluorescence Recovery After Photobleaching Gene Expression HeLa Cells Humans Luminescent Proteins/genetics,metabolism Mice Mice, Transgenic Microscopy, Fluorescence Olfactory Bulb/physiology Photic Stimulation Retina/metabolism Signal Transduction/physiology Transfection
Chemicals
Calcium Channels Luminescent Proteins Doxycycline Calcium
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Hasan Mazahir T
Max Planck Institute for Medical Research, Heidelberg, Germany. [email protected]
Friedrich Rainer W
Euler Thomas
Larkum Matthew E
Giese Günter
Both Matthias
Duebel Jens
Waters Jack
Bujard Hermann
Griesbeck Oliver
Tsien Roger Y
Nagai Takeharu
Miyawaki Atsushi
Denk Winfried
Conflict of Interest

The authors have declared that no conflicts of interest exist.

References (70)
70 references, click to expand
  1. Generating conditional mouse mutants via tetracycline-controlled gene expression.
    Methods Mol Biol. 2003;209:69-104 PMID: 12357963
  2. Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications.
    J Biol Chem. 2001 Aug 3;276(31):29188-94 PMID: 11387331
  3. Representation of odorants by receptor neuron input to the mouse olfactory bulb.
    Neuron. 2001 Nov 20;32(4):723-35 PMID: 11719211
  4. Genetically expressed cameleon in Drosophila melanogaster is used to visualize olfactory information in projection neurons.
    Curr Biol. 2002 Oct 29;12(21):1877-84 PMID: 12419190
  5. Two-photon laser scanning fluorescence microscopy.
    Science. 1990 Apr 6;248(4951):73-6 PMID: 2321027
  6. Differential regulation of active zone density during long-term strengthening of Drosophila neuromuscular junctions.
    J Neurosci. 2002 Nov 1;22(21):9399-409 PMID: 12417665
  7. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.
    Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547-51 PMID: 1319065
  8. Postsynaptic calcium is sufficient for potentiation of hippocampal synaptic transmission.
    Science. 1988 Oct 7;242(4875):81-4 PMID: 2845577
  9. Germ line transmission in transgenic mice.
    Prog Clin Biol Res. 1982;85 Pt B:111-24 PMID: 6181522
  10. Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector.
    Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8794-7 PMID: 1528894
  11. Spatially resolved calcium dynamics of mammalian Purkinje cells in cerebellar slice.
    Science. 1988 Nov 4;242(4879):773-7 PMID: 2847315
  12. Factors influencing frequency production of transgenic mice.
    Methods Enzymol. 1993;225:771-81 PMID: 8231885
  13. Mammalian cDNA and prokaryotic reporter sequences silence adjacent transgenes in transgenic mice.
    Nucleic Acids Res. 1997 Mar 1;25(5):1009-14 PMID: 9023112
  14. Two-photon calcium imaging reveals an odor-evoked map of activity in the fly brain.
    Cell. 2003 Jan 24;112(2):271-82 PMID: 12553914
  15. Novel fluorescent indicator proteins for monitoring free intracellular Ca2+.
    Cell Calcium. 1997 Sep;22(3):209-16 PMID: 9330791
  16. Dynamic and quantitative Ca2+ measurements using improved cameleons.
    Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2135-40 PMID: 10051607
  17. Detection of calcium transients in Drosophila mushroom body neurons with camgaroo reporters.
    J Neurosci. 2003 Jan 1;23(1):64-72 PMID: 12514202
  18. Genetically engineered mouse models of neurodegenerative diseases.
    Nat Neurosci. 2002 Jul;5(7):633-9 PMID: 12085093
  19. A GFP-equipped bidirectional expression module well suited for monitoring tetracycline-regulated gene expression in mouse.
    Nucleic Acids Res. 2001 Apr 1;29(7):E39 PMID: 11266574
  20. Photon upmanship: why multiphoton imaging is more than a gimmick.
    Neuron. 1997 Mar;18(3):351-7 PMID: 9115730
  21. Impermeant potential-sensitive oxonol dyes: III. The dependence of the absorption signal on membrane potential.
    J Membr Biol. 1988 Oct;105(1):55-64 PMID: 3225836
  22. Co-regulation of two gene activities by tetracycline via a bidirectional promoter.
    Nucleic Acids Res. 1995 Sep 11;23(17):3605-6 PMID: 7567477
  23. Synaptically activated Ca2+ waves in layer 2/3 and layer 5 rat neocortical pyramidal neurons.
    J Physiol. 2003 Jun 1;549(Pt 2):471-88 PMID: 12692172
  24. Light scattering and birefringence changes during nerve activity.
    Nature. 1968 May 4;218(5140):438-41 PMID: 5649693
  25. Role of ion flux in the control of c-fos expression.
    Nature. 1986 Aug 7-13;322(6079):552-5 PMID: 2426600
  26. Orchestrated response: a symphony of transcription factors for gene control.
    Genes Dev. 2000 Oct 15;14(20):2551-69 PMID: 11040209
  27. Genetically encoded indicators of cellular calcium dynamics based on troponin C and green fluorescent protein.
    J Biol Chem. 2004 Apr 2;279(14):14280-6 PMID: 14742421
  28. A high signal-to-noise Ca(2+) probe composed of a single green fluorescent protein.
    Nat Biotechnol. 2001 Feb;19(2):137-41 PMID: 11175727
  29. Ca2+-sensing transgenic mice: postsynaptic signaling in smooth muscle.
    J Biol Chem. 2004 May 14;279(20):21461-8 PMID: 14990564
  30. Imaging neuronal activity during zebrafish behavior with a genetically encoded calcium indicator.
    J Neurophysiol. 2003 Dec;90(6):3986-97 PMID: 12930818
  31. Directionally selective calcium signals in dendrites of starburst amacrine cells.
    Nature. 2002 Aug 22;418(6900):845-52 PMID: 12192402
  32. All-optical histology using ultrashort laser pulses.
    Neuron. 2003 Jul 3;39(1):27-41 PMID: 12848930
  33. Glutamate-induced intracellular acidification of cultured hippocampal neurons demonstrates altered energy metabolism resulting from Ca2+ loads.
    J Neurophysiol. 1994 Dec;72(6):2563-9 PMID: 7897473
  34. Long-term in vivo imaging of experience-dependent synaptic plasticity in adult cortex.
    Nature. 2002 Dec 19-26;420(6917):788-94 PMID: 12490942
  35. Optical methods for monitoring neuron activity.
    Annu Rev Neurosci. 1978;1:171-82 PMID: 386900
  36. Direct measurement of coupling between dendritic spines and shafts.
    Science. 1996 May 3;272(5262):716-9 PMID: 8614831
  37. A network of tufted layer 5 pyramidal neurons.
    Cereb Cortex. 1997 Sep;7(6):523-33 PMID: 9276177
  38. Long-term dendritic spine stability in the adult cortex.
    Nature. 2002 Dec 19-26;420(6917):812-6 PMID: 12490949
  39. Transcriptional activation by tetracyclines in mammalian cells.
    Science. 1995 Jun 23;268(5218):1766-9 PMID: 7792603
  40. In vivo two-photon calcium imaging of neuronal networks.
    Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7319-24 PMID: 12777621
  41. Studying gene function in eukaryotes by conditional gene inactivation.
    Annu Rev Genet. 2002;36:153-73 PMID: 12429690
  42. Circularly permuted green fluorescent proteins engineered to sense Ca2+.
    Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3197-202 PMID: 11248055
  43. Requirement for hippocampal CA3 NMDA receptors in associative memory recall.
    Science. 2002 Jul 12;297(5579):211-8 PMID: 12040087
  44. Patch-clamp recordings from the soma and dendrites of neurons in brain slices using infrared video microscopy.
    Pflugers Arch. 1993 Jun;423(5-6):511-8 PMID: 8351200
  45. Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity.
    Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):7963-8 PMID: 10859354
  46. DNA triplet repeats mediate heterochromatin-protein-1-sensitive variegated gene silencing.
    Nature. 2003 Apr 24;422(6934):909-13 PMID: 12712207
  47. Multi-neuronal signals from the retina: acquisition and analysis.
    J Neurosci Methods. 1994 Jan;51(1):95-106 PMID: 8189755
  48. Selective induction of LTP and LTD by postsynaptic [Ca2+]i elevation.
    J Neurophysiol. 1999 Feb;81(2):781-7 PMID: 10036277
  49. Changes in the intracellular concentration of free calcium ions in a pace-maker neurone, measured with the metallochromic indicator dye arsenazo III.
    J Physiol. 1978 Feb;275:357-76 PMID: 633127
  50. Intracellular signal transduction in four dimensions: from molecular design to physiology.
    Am J Physiol. 1992 Oct;263(4 Pt 1):C723-8 PMID: 1329539
  51. A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications.
    Nat Biotechnol. 2002 Jan;20(1):87-90 PMID: 11753368
  52. Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin.
    Nature. 1997 Aug 28;388(6645):882-7 PMID: 9278050
  53. Identification and function of thermosensory neurons in Drosophila larvae.
    Nat Neurosci. 2003 Mar;6(3):267-73 PMID: 12563263
  54. Long-term, noninvasive imaging of regulated gene expression in living mice.
    Genesis. 2001 Mar;29(3):116-22 PMID: 11252052
  55. Position-effect variegation and the genetic dissection of chromatin regulation in Drosophila.
    Semin Cell Dev Biol. 2003 Feb;14(1):67-75 PMID: 12524009
  56. Optical imaging of calcium transients in neurons and pharyngeal muscle of C. elegans.
    Neuron. 2000 Jun;26(3):583-94 PMID: 10896155
  57. A miniature head-mounted two-photon microscope. high-resolution brain imaging in freely moving animals.
    Neuron. 2001 Sep 27;31(6):903-12 PMID: 11580892
  58. Real-time imaging of calcium influx in mammalian cerebellar Purkinje cells in vitro.
    Proc Natl Acad Sci U S A. 1990 Jul;87(13):5084-8 PMID: 1973300
  59. Spatio-temporal dynamics of odor representations in the mammalian olfactory bulb.
    Neuron. 2002 Apr 11;34(2):301-15 PMID: 11970871
  60. Position-dependent variegation of globin transgene expression in mice.
    Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5371-5 PMID: 7777514
  61. Supralinear Ca2+ influx into dendritic tufts of layer 2/3 neocortical pyramidal neurons in vitro and in vivo.
    J Neurosci. 2003 Sep 17;23(24):8558-67 PMID: 13679425
  62. GABA- and glutamate-activated channels in green fluorescent protein-tagged gonadotropin-releasing hormone neurons in transgenic mice.
    J Neurosci. 1999 Mar 15;19(6):2037-50 PMID: 10066257
  63. Physiology and anatomy of synaptic connections between thick tufted pyramidal neurones in the developing rat neocortex.
    J Physiol. 1997 Apr 15;500 ( Pt 2):409-40 PMID: 9147328
  64. Molecular dissection of hippocampal theta-burst pairing potentiation.
    Proc Natl Acad Sci U S A. 2002 May 28;99(11):7740-5 PMID: 12032353
  65. Spatial and temporal analysis of calcium-dependent electrical activity in guinea pig Purkinje cell dendrites.
    Proc R Soc Lond B Biol Sci. 1990 May 22;240(1297):173-85 PMID: 1972990
  66. In vivo imaging of C. elegans mechanosensory neurons demonstrates a specific role for the MEC-4 channel in the process of gentle touch sensation.
    Neuron. 2003 Sep 11;39(6):1005-17 PMID: 12971899
  67. Circular permutation and receptor insertion within green fluorescent proteins.
    Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11241-6 PMID: 10500161
  68. Morphological changes in dendritic spines associated with long-term synaptic plasticity.
    Annu Rev Neurosci. 2001;24:1071-89 PMID: 11520928
  69. Control of memory formation through regulated expression of a CaMKII transgene.
    Science. 1996 Dec 6;274(5293):1678-83 PMID: 8939850
  70. CaMKII regulates the frequency-response function of hippocampal synapses for the production of both LTD and LTP.
    Cell. 1995 Jun 16;81(6):891-904 PMID: 7781066
Article Info
Journal
PLoS biology
Abbr.
PLoS Biol
ISSN
1545-7885
DOI
10.1371/journal.pbio.0020163
Email
[email protected]
Published
2004-06-00
Epub
2004-00-15
Pages
e163
Language
English
Region
United States
NLM ID
101183755
PMCID
PMC423138
Subset
IM
Grants
NINDS NIH HHS · R01 NS027177 · United States
NINDS NIH HHS · R37 NS027177 · United States
NINDS NIH HHS · NS27177 · United States
External Links
PubMed source DOI: 10.1371/journal.pbio.0020163 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2