Hindbrain and Cranial Nerve Dysmorphogenesis Result from Acute Maternal Ethanol Administration

Dunty, William C.; Zucker, Robert M.; Sulik, Kathleen K.

doi:10.1159/000066748

Cited by 65 publications

(94 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have previously demonstrated that apoptosis (cell death) can be visualized in 8-9 days old mouse embryos, rat fetal limbs, and other tissues using a fluorescent dye (LysoTracker Red, LT), which stains lysosomes and acidic regions of cells/tissues (16)(17)(18)(19)(20)(21)(22)(23). The current study extends our previous work to visualize apoptosis within follicles of a whole ovary (17).…”

supporting

confidence: 66%

“…From previous studies on embryos, the appearance of LT in tissue is strongly suggestive of apoptosis (16)(17)(18)(19)(20)(21)(22)(23). The LT has a fluorescence peak at 610 nm (Leica SP1), while the AIF has a fluorescence peak around 471 nm with UV or cyan GFP excitation.…”

Section: Discussionmentioning

confidence: 92%

“…In the case of the ovary, the LT was excited with a 543 nm laser to indicate the regions of specific apoptosis that existed in the tissue (16)(17)(18)(19)(20)(21)(22)(23) while the background fluorescence consisting primarily of aldehyde autofluorescence that was optimally excited using the 488 nm laser. The combination of two lasers achieved the specificity of the LT probe for apoptotic cells and the morphology of the tissue was achieved by the Glut probe.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Confocal laser scanning microscopy of whole mouse ovaries: Excellent morphology, apoptosis detection, and spectroscopy

Zucker

Jeffay

2006

Cytometry Pt A

Self Cite

View full text Add to dashboard Cite

Background:Ovaries consist of numerous follicles, oocytes, and granulosa cells in different stages of development. Many of these follicles will undergo an apoptotic process during the lifetime of the animal. By using proper tissue preparation methods, the events within the whole ovary can be observed by using 3D confocal microscopy.Methods:Whole ovaries were stained with LysoTracker Red (LT), fixed with 4% paraformaldehyde (PF) and 1% glutaraldehyde (Glut), dehydrated with methanol (MEOH), and cleared with benzyl alcohol and benzyl benzoate (BABB). Using this tissue preparation technique, the ovary becomes relatively transparent, allowing its morphology to be observed with confocal microscopes. A spectral imaging system (PARISS) located on a conventional microscope was used to interpret the LT dye spectra and fixation products in the tissues with different excitation wavelengths.Results:Apoptosis in the follicle was detected as clusters of intensely stained granulosa cells located in close proximity to the oocytes. The fixation with Glut and PF preserved morphological details, increased tissue fluorescence, thus increased the signal to noise of the background image.Conclusions:Thick tissues can be imaged after they are properly stained, aldehyde fixed, and BABB cleared. LT intensely stained single cells or clusters of apoptotic cells in the follicles and the nucleolus. Spectral differences between LT as an indicator of apoptosis and Glut‐PF fixation was used to visualize ovarian morphology and apoptosis. The PARISS spectrophotometer revealed spectral peaks for LT at 609.6 nm and for Glut‐PF at 471.3 nm. The proper use of the spectra from these fluorescence molecules is the foundation for high quality morphological images of apoptosis. By sequentially imaging the two probes with a 488 nm laser and a 543/568 nm laser, there was a reduction in fluorescent cross talk and an increase in image quality. © 2006 International Society for Analytical Cytology

show abstract

supporting

confidence: 66%

Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Confocal laser scanning microscopy of whole mouse ovaries: Excellent morphology, apoptosis detection, and spectroscopy

Zucker

Jeffay

2006

Cytometry Pt A

Self Cite

View full text Add to dashboard Cite

show abstract

“…Four nuclei affected by ethanol exposure during gastrulation include three trigeminal nuclei (the PSN, SpVo, and MoV) and the MoVII. These changes in the brainstem occur after an exposure to ethanol that is associated with FAS-like craniofacial malformations (Sulik et al, 1981;Astley et al, 1999;Dunty et al, 2002;Da Lee et al, 2004). In addition, four nuclei (the MoV, MoVII, MoXII, and PSN) are affected by ethanol exposure during neuronal generation.…”

Section: Timing Of the Ethanol-induced Vulnerabilitymentioning

confidence: 99%

“…One critical window of ethanol vulnerability coincides with the period of gastrulation. In mice, exposure to ethanol on gestational day (G) 7 or G8 increases the expression of markers for cell death (Dunty et al, 2002;Kilburn et al, 2006). More impressive is that mice exposed to ethanol during gastrulation (Sulik et al, 1981;Dunty et al, 2002;Da Lee et al, 2004) exhibit craniofacial malformations like those that characterize human children with fetal alcohol syndrome (FAS) (Lemoine et al, 1968;Jones and Smith, 1973).…”

Section: Introductionmentioning

confidence: 99%

Time-specific effects of ethanol exposure on cranial nerve nuclei: Gastrulation and neuronogenesis

Mooney

Miller

2007

Experimental Neurology

View full text Add to dashboard Cite

During the development of the central nervous system, neurons pass through critical periods or periods of vulnerability. We explored periods of vulnerability for cranial nerve nuclei by determining the effects of acute exposure to ethanol during development on the number of neurons in mature brainstem. Long-Evans rats were injected with 2.9 g ethanol/kg body weight on one day between gestational day (G) 7 and G13, inclusive. Two hours later, animals received a second injection of 1.45 g/kg. Controls were injected with equivalent volumes of saline. Brainstems of 31-day-old offspring were cryosectioned and stained with cresyl violet. Stereological methods were used to determine the volume and numerical density of neurons in three trigeminal sensory nuclei (the principal sensory nucleus of the trigeminal nerve, and the oral and interpolar subnuclei of the spinal trigeminal nuclear complex) and three motor nuclei (the trigeminal, facial, and hypoglossal nuclei). The numbers of neurons in most nuclei were lower following early (on G7 and/or G8) or later (on G12 and/or G13) exposure. Only the trigeminal interpolar nucleus was affected by neither early nor late ethanol exposure. Thus, prenatal exposure to ethanol affects the number of neurons in brainstem nuclei in a time-dependent manner. Windows of vulnerability coincide with gastrulation (G7/G8) and the period of neuronal generation (G12/G13).

show abstract

The implications of biologic therapy in ankylosing spondylitis: Comment on the articles by Brandt et al and Braun et al

Mascarenhas¹,

Davis²,

Robertson³

2004

Arthritis & Rheumatism

View full text Add to dashboard Cite

Background: After fluorochromes are incorporated into cells, tissues, and organisms, confocal microscopy can be used to observe three‐dimensional structures. LysoTracker Red (LT) is a paraformaldehyde fixable probe that concentrates into acidic compartments of cells and indicates regions of high lysosomal activity and phagocytosis, which both correlate to apoptosis activity. LT has been shown to be an indicator of apoptotic cell death which is correlated to other standard apoptotic assays. Methods: The mammalian samples were stained with LT, fixed with paraformaldehyde/glutaraldehyde, dehydrated with methanol (MEOH), and cleared with benzyl alcohol/benzyl benzoate (BABB). Following this treatment, the tissues were nearly transparent. Mosquitoes were fixed with MEOH and stained with propidium iodide. Next the tissues were dehydrated with MEOH and cleared with BABB. Results: Tissues as thick as 500 μm can be visualized after clearing with BABB. LT staining revealed apoptotic regions in mammalian limbs, fetuses, and embryos. Morphological observation of insect tissue consisted of combining autofluorescence with either nucleic acid staining (either propidium iodide or ethidium bromide). Conclusions: The use of BABB matches the RI of the tissue within the suspending medium. It helps in increasing the penetration of laser light in a confocal microscope by reducing the amount of light scattering artifacts and allows for the visualization of morphology in thick tissues. LT is a probe that stains the acid regions of tissues and cells and has been correlated to apoptosis. Morphological features of a tissue or organism (embryo, mosquito larvae) can be elucidated by fixation aldehydes, autofluorescence, and red‐emitting probes. This sample preparation procedure with optimization of confocal laser scanning microscopy allowed for the detection and visualization of apoptosis in fetal limbs and embryos which were ∼500‐μm thick. © 2006 International Society for Analytical Cytology

show abstract

Hindbrain and Cranial Nerve Dysmorphogenesis Result from Acute Maternal Ethanol Administration

Cited by 65 publications

References 60 publications

Confocal laser scanning microscopy of whole mouse ovaries: Excellent morphology, apoptosis detection, and spectroscopy

Confocal laser scanning microscopy of whole mouse ovaries: Excellent morphology, apoptosis detection, and spectroscopy

Time-specific effects of ethanol exposure on cranial nerve nuclei: Gastrulation and neuronogenesis

The implications of biologic therapy in ankylosing spondylitis: Comment on the articles by Brandt et al and Braun et al

Contact Info

Product

Resources

About