A two focal plane method for digital quantification of nuclear immunoreactivity in large brain areas using NIH-image software

Brown, Harold E.; Garcia, Meredith M.; Harlan, Richard E.

doi:10.1016/s1385-299x(98)00003-8

Cited by 40 publications

(33 citation statements)

References 17 publications

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“…1.61). 38 Surgery and microdialysis Mice were anesthetized with pentobarbital (50 mg kg -1 , Somnopentyl, Schering Plough Animal Health, Kenilworth, NJ, USA) and mounted on a stereotaxic frame. Stainless guide cannula (AG-4, Eicom, Kyoto, Japan) with a dummy probe (AD-4, Eicom) was placed in the medial prefrontal cortex (mpFC, equivalent to prelimbic cortex) (coordinates: anterior þ 2.0 mm, lateral þ 0.5 mm, ventral À6.0 mm relative to the bregma).…”

Section: Methamphetamine Challengementioning

confidence: 99%

Transient exposure of neonatal mice to neuregulin-1 results in hyperdopaminergic states in adulthood: implication in neurodevelopmental hypothesis for schizophrenia

et al. 2010

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Neuregulin-1 (NRG1) is implicated in the etiology or pathology of schizophrenia, although its biological roles in this illness are not fully understood. Human midbrain dopaminergic neurons highly express NRG1 receptors (ErbB4). To test its neuropathological role in the neurodevelopmental hypothesis of schizophrenia, we administered type-1 NRG1 protein to neonatal mice and evaluated the immediate and subsequent effects on dopaminergic neurons and their associated behaviors. Peripheral NRG1 administration activated midbrain ErbB4 and elevated the expression, phosphorylation and enzyme activity of tyrosine hydroxylase (TH), which ultimately increased dopamine levels. The hyperdopaminergic state was sustained in the medial prefrontal cortex after puberty. There were marked increases in dopaminergic terminals and TH levels. In agreement, higher amounts of dopamine were released from this brain region of NRG1-treated mice following high potassium stimulation. Furthermore, NRG1-treated mice exhibited behavioral impairments in prepulse inhibition, latent inhibition, social behaviors and hypersensitivity to methamphetamine. However, there were no gross abnormalities in brain structures or other phenotypic features of neurons and glial cells. Collectively, our findings provide novel insights into neurotrophic contribution of NRG1 to dopaminergic maldevelopment and schizophrenia pathogenesis.

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Section: Methamphetamine Challengementioning

confidence: 99%

Transient exposure of neonatal mice to neuregulin-1 results in hyperdopaminergic states in adulthood: implication in neurodevelopmental hypothesis for schizophrenia

et al. 2010

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“…Of particular interest is the fact that color separation is effectively performed at both low ( Figure 2) and high magnifications (Figure 2, insets). In contrast to RGB thresholding techniques, which demonstrate improved separation capacities at higher magnifications (Brown et al 1998), Photoshop-based image analysis uses the color information contained in each individual pixel (color hue, saturation, luminosity) and thus allows separation of colors at the levels of pixel resolution. This also makes it possible to completely separate chromogens with closely related spectral characteristics, which is particularly relevant to the separation of brown and blue for the separation of the immunoperoxidase signal from the hematoxylin counterstain.…”

Section: Discussionmentioning

confidence: 99%

Complete Chromogen Separation and Analysis in Double Immunohistochemical Stains Using Photoshop-based Image Analysis

Lehr

Loss

Teeling

et al. 1999

J Histochem Cytochem.

203

153

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SUMMARYSimultaneous detection of two different antigens on paraffin-embedded and frozen tissues can be accomplished by double immunohistochemistry. However, many double chromogen systems suffer from signal overlap, precluding definite signal quantification. To separate and quantitatively analyze the different chromogens, we imported images into a Macintosh computer using a CCD camera attached to a diagnostic microscope and used Photoshop software for the recognition, selection, and separation of colors. We show here that Photoshop-based image analysis allows complete separation of chromogens not only on the basis of their RGB spectral characteristics, but also on the basis of information concerning saturation, hue, and luminosity intrinsic to the digitized images. We demonstrate that Photoshop-based image analysis provides superior results compared to color separation using bandpass filters. Quantification of the individual chromogens is then provided by Photoshop using the Histogram command, which supplies information on the luminosity (corresponding to gray levels of black-and-white images) and on the number of pixels as a measure of spatial distribution. (J Histochem Cytochem 47:119-125, 1999)

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“…Quantification of immunoreactive cells was based on methods previously described (Brown et al 1998;Countryman and Gold 2007). Sections were imaged using a Leica DM 6000B/CTR6000 light microscope and a Leica DFC350 FX video camera, which was interfaced to an EP PC computer.…”

Section: Quantification Of Creb Pcreb and C-fos Immunoreactive Cellsmentioning

confidence: 99%

Intra-amygdala injections of CREB antisense impair inhibitory avoidance memory: Role of norepinephrine and acetylcholine

2008

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Infusions of CREB antisense into the amygdala prior to training impair memory for aversive tasks, suggesting that the antisense may interfere with CRE-mediated gene transcription and protein synthesis important for the formation of new memories within the amygdala. However, the amygdala also appears to modulate memory formation in distributed brain sites, through mechanisms that include the release of norepinephrine and acetylcholine within the amygdala. Thus, CREB antisense injections may affect memory by interfering with mechanisms of modulation, rather than storage, of memory. In the present experiment, rats received bilateral intra-amygdala infusions of CREB antisense (2 nmol/1 µL) 6 h prior to inhibitory avoidance training. In vivo microdialysis samples were collected from the right amygdala before, during, and following training. CREB antisense produced amnesia tested at 48 h after training. In addition, CREB antisense infusions dampened the training-related release of norepinephrine, and to a lesser extent of acetylcholine, in the amygdala. Furthermore, intra-amygdala infusions of the ␤-adrenergic receptor agonist clenbuterol administered immediately after training attenuated memory impairments induced by intra-amygdala injections of CREB antisense. These findings suggest that intra-amygdala treatment with CREB antisense may affect processes involved in modulation of memory in part through interference with norepinephrine and acetylcholine neurotransmission in the amygdala.

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A two focal plane method for digital quantification of nuclear immunoreactivity in large brain areas using NIH-image software

Cited by 40 publications

References 17 publications

Transient exposure of neonatal mice to neuregulin-1 results in hyperdopaminergic states in adulthood: implication in neurodevelopmental hypothesis for schizophrenia

Transient exposure of neonatal mice to neuregulin-1 results in hyperdopaminergic states in adulthood: implication in neurodevelopmental hypothesis for schizophrenia

Complete Chromogen Separation and Analysis in Double Immunohistochemical Stains Using Photoshop-based Image Analysis

Intra-amygdala injections of CREB antisense impair inhibitory avoidance memory: Role of norepinephrine and acetylcholine

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