Electron Microscopic Preembedding Double-Immunostaining Methods

Leranth, Csaba; Pickel, Virginia M.

doi:10.1007/978-1-4757-2055-6_6

Cited by 197 publications

(247 citation statements)

References 87 publications

Supporting

Mentioning

242

Contrasting

Order By: Relevance

“…The use of each antiserum at optimal high dilutions significantly reduced background labeling, enabling identification of relatively low levels and heterogeneously dispersed aggregates of immunoreactivity within the cytoplasm. The immunogold labeling method, however, is considerably less sensitive than the avidin-biotin-peroxidase method (Leranth and Pickel, 1989). The differences in sensitivity, together with the heterogeneous cytoplasmic and plasmalemmal distribution of both AT 1 receptor and gp91 phox , may have resulted in an underestimation of the number of profiles that were single or dually labeled.…”

Section: Methodological Considerationsmentioning

confidence: 99%

NADPH Oxidase Contributes to Angiotensin II Signaling in the Nucleus Tractus Solitarius

Wang¹,

Anrather²,

Huang³

et al. 2004

J. Neurosci.

157

183

View full text Add to dashboard Cite

Section: Methodological Considerationsmentioning

confidence: 99%

NADPH Oxidase Contributes to Angiotensin II Signaling in the Nucleus Tractus Solitarius

Wang¹,

Anrather²,

Huang³

et al. 2004

J. Neurosci.

157

183

View full text Add to dashboard Cite

“…They were washed in PB, dehydrated through a series of increasing alcohol concentrations, and incubated overnight in a 1:1 mixture of propylene oxide and Epon 812. They were then flat-embedded in Epon 812 (Leranth and Pickel, 1989), mounted on Epon blocks, and thin-sectioned with an LKB ultramicrotome using a diamond knife (Diatome, Fort Washington, PA). Thin sections of the shell region of the Acb were taken from the outer surface of the tissue.…”

Section: Methodsmentioning

confidence: 99%

Dual Ultrastructural Localization of μ-Opioid Receptors and NMDA-Type Glutamate Receptors in the Shell of the Rat Nucleus Accumbens

1997

Self Cite

View full text Add to dashboard Cite

The effectiveness of NMDA antagonists in modulating the motor and motivational effects of opiates is attributed, in part, to functional associations involving NMDA receptors and -opioid receptors (MORs) in the shell of the nucleus accumbens (Acb). To determine the subcellular sites for potential functional interactions between opiate ligands and NMDA receptors in this region, we examined the ultrastructural localization of antipeptide antisera against MOR and the R1 subunit of the NMDA receptor in the Acb shell of the adult rat brain. MOR-like immunoreactivity (MOR-LI) was seen primarily in dendrites, whereas NMDAR1-like immunoreactivity (NMDAR1-LI) was detected more often in axon terminals forming asymmetric synapses. In these profiles, MOR labeling was localized mainly to extrasynaptic plasma membranes, whereas NMDAR1-LI was associated with both synaptic and extrasynaptic sites. Of 307 MOR-labeled processes, 17.9% of the dendrites and 9.4% of the axon terminals also contained NMDAR1-LI. In addition, 24.7% of the dendrites containing only MOR-LI were apposed to NMDAR1-labeled axons or terminals. We conclude that in the shell of the Acb, the output of single neurons can be dually modulated by (1) activation of MOR and NMDA receptors in the same dendrites or (2) combined activation of presynaptic NMDA receptors in afferents contacting dendrites containing MOR. In addition, the colocalization of MOR and NMDAR1 in certain axon terminals in the Acb suggests their dual involvement in the presynaptic release of neurotransmitters in this region.

show abstract

“…Mouse brains were prepared for electron microscopy as previously described (Leranth and Pickel, 1989;Chan et al, 1990). Briefly, mice were deeply anesthetized with sodium pentobarbital (150 mg/kg, i.p.)…”

Section: Tissue Preparation For Electron Microscopymentioning

confidence: 99%

Conditional deletion of the NMDA-NR1 receptor subunit gene in the central nucleus of the amygdala inhibits naloxone-induced conditioned place aversion in morphine-dependent mice

Glass

Hegarty²,

Oselkin

et al. 2008

Experimental Neurology

Self Cite

View full text Add to dashboard Cite

Preclinical behavioral pharmacological and neuropharmacological evidence indicates that the NMDA receptor plays an important role in opioid dependence, however, the neural substrates subserving these actions are poorly understood. The central nucleus of the amygdala (CeA) is a critical coordinator of autonomic, behavioral, and emotional systems impacted by opioids, however there is no evidence that the essential NMDA-NR1 (NR1) subunit gene in the amygdala plays a role in opioid dependence. To determine the role of the NR1 subunit gene in the amygdala with respect to physical and psychological opioid withdrawal, a spatial-temporal deletion of this gene was produced by microinjecting a recombinant adeno-associated virus (rAAV) expressing the GFP reporter and Cre recombinase (rAAV-GFP-Cre) into the CeA of adult "floxed" NR1 mice (fNR1). Amygdala microinjection of rAAV-GFP-Cre produced a decrease in NR1 gene expression and protein immunolabeling in postsynaptic sites of neurons without signs of compromised ultrastructural neuronal morphology. Amygdala NR1 gene deletion also did not affect locomotor, somatosensory, or sensory-motor behaviors. In addition, bilateral local NR1 gene deletion did not impact somatic or visceral withdrawal symptoms precipitated by naloxone in morphine-dependent mice. However, there was a significant deficit in the expression of an opioid withdrawal-induced conditioned place aversion in mice with amygdala NR1 deletion. These results indicate that functional amygdala NMDA receptors are involved in aversive psychological Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access

show abstract

Electron Microscopic Preembedding Double-Immunostaining Methods

Cited by 197 publications

References 87 publications

NADPH Oxidase Contributes to Angiotensin II Signaling in the Nucleus Tractus Solitarius

NADPH Oxidase Contributes to Angiotensin II Signaling in the Nucleus Tractus Solitarius

Dual Ultrastructural Localization of μ-Opioid Receptors and NMDA-Type Glutamate Receptors in the Shell of the Rat Nucleus Accumbens

Conditional deletion of the NMDA-NR1 receptor subunit gene in the central nucleus of the amygdala inhibits naloxone-induced conditioned place aversion in morphine-dependent mice

Contact Info

Product

Resources

About