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Cell- and lamina-specific expression and activity-dependent regulation of type II calcium/calmodulin-dependent protein kinase isoforms in monkey visual cortex - PubMed

  • ️Thu Jan 01 1998

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Cell- and lamina-specific expression and activity-dependent regulation of type II calcium/calmodulin-dependent protein kinase isoforms in monkey visual cortex

B Tighilet et al. J Neurosci. 1998.

Abstract

In situ hybridization histochemistry and immunocytochemistry were used to study localization and activity-dependent regulation of alpha, beta, gamma, and delta isoforms of type II calcium/calmodulin-dependent protein kinase (CaMKII) and their mRNAs in areas 17 and 18 of normal and monocularly deprived adult macaques. CaMKII-alpha is expressed overall at levels three to four times higher than that of CaMKII-beta and at least 15 times higher than that of CaMKII-gamma and -delta. All isoforms are expressed primarily in pyramidal cells of both areas, especially those of layers II-III, IVA (in area 17), and VI, but are also expressed in nonpyramidal, non-GABAergic cells of layer IV of both areas and in interstitial neurons of the white matter. CaMKII-alpha and -beta are colocalized, suggesting the formation of heteromers. There was no evidence of expression in neuroglial cells. Each isoform has a unique pattern of laminar and sublaminar distribution, but cortical layers or sublayers enriched for one isoform do not correlate with layers receiving inputs only from isoform-specific layers of the lateral geniculate nucleus. CaMKII-alpha and -beta mRNA and protein levels in layer IVC of area 17 are subject to activity-dependent regulation, with brief periods of monocular deprivation caused by intraocular injections of tetrodotoxin leading to a 30% increase in CaMKII-alpha mRNA and a comparable decrease in CaMKII-beta mRNA in deprived ocular dominance columns, especially of layer IVCbeta. Expression in other layers and expression of CaMKII-gamma and delta were unaffected. Changes occurring in layer IVC may influence the formation of heteromers and protect supragranular layers from CaMKII-dependent plasticity in the adult.

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Figures

Fig. 1.
Fig. 1.

Autoradiograms of surface parallel sections near the occipital pole of the same M. fuscata brain that were hybridized to radioactive RNA probes complementary to CaMKII-α (CAM-α), CaMKII-β (CAM-β), CaMKII-γ (CAM-γ), and CaMKII-δ (CAM-δ) mRNAs. A is more superficial than B–D which are closely adjacent to one another.Arrows indicate the border between areas 17 and 18.A was exposed for 3 d, B was exposed for 5 d, and C and D were exposed for 11 d to reveal laminar densities of hybridization. For true relative differences in overall densities of expression, see Figure 2. Scale bar, 1 mm.

Fig. 2.
Fig. 2.

Integrated optical density readings (IOD) converted to measures of radioactivity (nCi/gm) by reference to standards exposed on the same sheet of film and taken in traverses of constant width across the thickness of areas 17 and 18 in autoradiograms similar to those of Figure 1 but exposed for the same time on the same sheet of film. These reveal laminar patterns of expression as well as relative levels of expression of the four CaMKII isoforms. Note that the scales of the y axes differ.Roman numerals on the x axes of thetop graphs represent positions of cortical layers in all scans, as determined from optical density readings of adjacent Nissl-stained sections.

Fig. 3.
Fig. 3.

Pairs of dark-field (left) and bright-field (right) photomicrographs from the same counterstained emulsion autoradiograms showing relative densities and laminar patterns of expression of the four CaMKII mRNAs in areas 17 (A, C, E,G) and 18 (B, D,F, H). More diffuse labeling for CaMKII-α, tending to obscure borders between layers at this magnification, reflects the high levels of the mRNA in dendrites of pyramidal cells (Benson et al., 1991a). Scale bar, 100 μm.

Fig. 4.
Fig. 4.

High-magnification photomicrographs from emulsion autoradiograms showing hybridization of cRNAs specific for each of the CaMKII mRNAs to neurons in layer VI of area 17 (A), layer V of area 18 (B), layer III of area 18 (C), and layer VI of area 17 (D). Arrows indicate nuclei of neuroglial cells which are not labeled. Scale bar, 10 μm.

Fig. 5.
Fig. 5.

Photomicrographs from pairs of adjacent sections stained immunocytochemically (left) for one of the four CaMKII isoforms or with thionin (right) and showing the laminar patterns of immunostaining in areas 17 (A,C, E, G) and 18 (B, D, F,H). Dense neuropil staining in the absence of somal staining (e.g., layers IVA and V of area 17) reflects immunostaining of axons and dendrites. Scale bar, 100 μm.

Fig. 6.
Fig. 6.

High-magnification photomicrographs showing immunostaining of cells in different layers of areas 17 or 18 for CaMKII-α (A–E) or CaMKII-β (F, G). (These are different sections from those shown in Fig. 5.) Note the staining of pyramidal cells in all layers and the staining of small round cell somata in layers IVC (area 17) and IV (area 18). WM, White matter. Scale bar, 150 μm.

Fig. 7.
Fig. 7.

A–D, Bright-field photomicrographs showing immunostaining of cells for CaMKII-γ or CaMKII-δ in deeper layers of areas 17 and 18. The isolated large neuronal somata stained in layer V of area 17 are those of Meynert cells. Scale bars, 150 μm.E, F, Fluorescence micrographs from the same microscopic field showing double staining of cells in layer III of area 17 for CaMKII-α (E, rhodamine immunofluorescence) and CaMKII-β (F, fluorescein immunofluorescence). Scale bar, 10 μm.

Fig. 8.
Fig. 8.

Paired fluorescence photomicrographs from the same microscopic fields stained for CaMKII-α (CAM-α) and GABA (A–D) or for CaMKII-β (CAM-β) and GABA (E–H).A, B, E, F, From layer III of area 17 and showing CaMKII-α and -β immunoreactivity in pyramidal cells but not in GABA cells.C, D, G, H, From layer IVCβ of area 17 and showing CaMKII-α and -β immunoreactivity in small, presumably spiny stellate cells and not in GABA cells. In the CaMKII-α- or CaMKII-β-immunostained member of each pair of micrographs, the unstained GABA cells are indicated byasterisks. Scale bar, 10 μm.

Fig. 9.
Fig. 9.

Pairs (A, B;C, D) of adjacent sections from area 17 stained immunocytochemically for CaMKII-α (A) or CaMKII-β (C) or for CO (B,D). Sections are from a M. fascicularis(A, B) and a M. fuscata(C, D) monkey subjected to monocular TTX injections for 7 d. Zones of reduced CO staining in layer IVC represent ocular dominance columns related to the deprived eye. CaMKII-α immunostaining is enhanced and CaMKII-β immunostaining is reduced in regions corresponding to the deprived columns, especially in layer IVCβ. Arrows in A andB and circles in C andD indicate the same blood vessels. Scale bars:A, B, 250 μm; C,D, 500 μm.

Fig. 10.
Fig. 10.

A, Autoradiogram from a surface parallel section through area 17 of a M. fuscata monkey monocularly deprived for 7 d. The section was hybridized with a CaMKII-α riboprobe. B, An adjacent section stained for CO. C, D, Enlargements of upper parts of A and B.Circles indicate the same blood vessels. Scale bars:A, B, 2 mm; C,D, 1 mm. E, F, Optical density scans made across layer IVC in regions indicated by thelines between the arrows inC and D converted to measures of radioactivity to show enhancement of CaMKII-α mRNA levels in deprived ocular dominance columns, the positions of which can be determined by matching to the zones showing reduced CO staining and lowered optical density.

Fig. 11.
Fig. 11.

A, Autoradiogram from a surface parallel section through area 17 of a M. fuscata monkey monocularly deprived for 16 d. The section was hybridized with a CaMKII-β riboprobe. B, An adjacent section stained for CO. C, D, Enlargements of middle regions of A and B.Circles indicate the same blood vessels, andarrows indicate the same deprived ocular dominance columns in layer IVC. E, F, Optical density scans made along the lines indicated inC and D and showing decreased CaMKII-β mRNA levels in the deprived ocular dominance columns. Scale bars:A, B, 2 mm; C,D, 500 μm.

Fig. 12.
Fig. 12.

A, C, Autoradiograms of sections through area 17 of a M. fuscata monkey monocularly deprived for 7 d.A, Section hybridized to a CaMKII-γ riboprobe.C, Section hybridized to a CaMKII-δ riboprobe. Sections have been exposed for 15 d. B,D, The adjacent CO-stained sections. No deprivation effect can be detected for either mRNA. Scale bar, 2 mm.

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References

    1. Artola A, Singer W. Long-term potentiation and NMDA receptors in rat visual cortex. Nature. 1987;330:649–652. - PubMed
    1. Artola A, Brocher S, Singer W. Different voltage-dependent thresholds for inducing long-term depression and long-term potentiation in slices of rat visual cortex. Nature. 1990;347:69–72. - PubMed
    1. Bear MF, Press WA, Connors BW. Long-term potentiation in slices of kitten visual cortex and the effects of NMDA receptor blockade. J Neurophysiol. 1992;67:841–851. - PubMed
    1. Bennett MK, Kennedy MB. Deduced primary structure of the β subunit of brain type II Ca2+/calmodulin-dependent protein kinase determined by molecular cloning. Proc Natl Acad Sci USA. 1987;84:1794–1798. - PMC - PubMed
    1. Bennett MK, Erondu NE, Kennedy MB. Purification and characterization of a calmodulin-dependent protein kinase that is highly concentrated in brain. J Biol Chem. 1983;258:12735–12744. - PubMed

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