Immunocytochemistry of endothelial nitric oxide synthase in the rat brain: a light and electron microscopical study using the tyramide signal amplification technique

Stanarius, Andreas; Töpel, Ingolf; Schulz, Stefan; Noack, Heiko; Wolf, Gerald

doi:10.1016/s0065-1281(97)80034-7

Cited by 85 publications

(46 citation statements)

References 72 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the present study, we also found a parallel increase in NOS activity in cerebral vessels isolated from estrogen-treated animals. Intraparenchymal vessels contain only eNOS, but pial arteries can also contain perivascular neuronal NOS (nNOS)-positive nerves 26 ; therefore, we confirmed our NOS activity results using vessels isolated from brains in which the pial vessels had been removed. Furthermore, preliminary studies in our laboratory indicate that estrogen has no effect on nNOS protein (K.J.…”

Section: Discussionsupporting

confidence: 79%

Estrogen Increases Endothelial Nitric Oxide Synthase via Estrogen Receptors in Rat Cerebral Blood Vessels

McNeill

Zhang

Stanczyk

et al. 2002

Stroke

134

View full text Add to dashboard Cite

Background and Purpose-In vivo and in vitro rat models of hormone therapy were used to test the following hypotheses:(1) estrogen acts directly on cerebrovascular estrogen receptors to increase endothelial nitric oxide synthase (eNOS); (2) increased protein correlates with higher NOS activity; and (3) effects of estrogen on eNOS are altered by concurrent treatment with either medroxyprogesterone acetate (MPA) or progesterone. Methods-Blood vessels were isolated from brains of ovariectomized female rats; some were treated for 1 month with estrogen, estrogen and progesterone, or estrogen and MPA. The latter effect persists in the presence of either progesterone or MPA. Thus, increased NO production by eNOS may contribute to the neuroprotective effects of estrogen.

show abstract

Section: Discussionsupporting

confidence: 79%

Estrogen Increases Endothelial Nitric Oxide Synthase via Estrogen Receptors in Rat Cerebral Blood Vessels

McNeill

Zhang

Stanczyk

et al. 2002

Stroke

134

View full text Add to dashboard Cite

show abstract

“…It was originally reported to be in CA1 pyramidal cells . However, subsequent studies by the same laboratory (Demas et al, 1999;Blackshaw et al, 2003) and others (Seidel et al, 1997;Stanarius et al, 1997;Topel et al, 1998) could not confirm this result and, instead, concluded that eNOS mRNA and protein were found only in endothelial cells. Hence, the tonic NO we measured is likely to come from this source.…”

Section: Discussionmentioning

confidence: 92%

Tonic and Phasic Nitric Oxide Signals in Hippocampal Long-Term Potentiation

Hopper¹,

Garthwaite²

2006

J. Neurosci.

199

150

View full text Add to dashboard Cite

Nitric oxide (NO) participates in long-term potentiation (LTP) and other forms of synaptic plasticity in many different brain areas but where it comes from and how it acts remain controversial. Using rat and mouse hippocampal slices, we tested the hypothesis that tonic and phasic NO signals are needed and that they derive from different NO synthase isoforms. NMDA increased NO production in a manner that was potently inhibited by three different neuronal NO synthase (nNOS) inhibitors. Tonic NO could be monitored after sensitizing guanylyl cyclase-coupled NO receptors, allowing the very low ambient NO concentrations to be detected by cGMP measurement. The levels were unaffected by inhibition of NMDA receptors, nNOS, or the inducible NO synthase (iNOS). iNOS was also undetectable in protein or activity assays. Tonic NO was susceptible to agents inhibiting endothelial NO synthase (eNOS) and was missing in eNOS knock-out mice. The eNOS knock-outs exhibited a deficiency in LTP resembling that seen in wild-types treated with a NO synthase inhibitor. LTP in the knock-outs could be fully restored by supplying a low level of NO exogenously. Inhibition of nNOS also caused a major loss of LTP, particularly of late-LTP. Again, exogenous NO could compensate, but higher concentrations were needed compared with those restoring LTP in the eNOS knock-outs. It is concluded that tonic and phasic NO signals are both required for hippocampal LTP and the two are generated, respectively, by eNOS and nNOS, the former in blood vessels and the latter in neurons.

show abstract

“…Soon after the technique was first described, its widespread utilization (Van Heusden et al 1997;Kaufmann et al 1998;Kressel 1998;van de Corput et al 1998;Speel et al 1998Speel et al ,1999 and further modifications in its application highlighted its usefulness and importance, particularly in paradigms involving fluorescent double labeling with antibodies derived from the same species (Hunyady et al 1996;Shindler and Roth 1996;Teramoto et al 1998). Recently, this technique has also been used for pre-and postembedding immunoelectron microscopy, an extremely labor-intensive approach (Schofer et al 1997;Stanarius et al 1997;Punnonen et al 1999), with some suggesting that the electron-dense tyramide can also be used for EM without concomitant DAB chromogen application (Mayer and Bendayan 1997).…”

Section: Discussionmentioning

confidence: 99%

Novel Application of Tyramide Signal Amplification (TSA): Ultrastructural Visualization of Double-labeled Immunofluorescent Axonal Profiles

Büki

Walker

Stone

et al. 2000

J Histochem Cytochem.

View full text Add to dashboard Cite

SUMMARY Fluorescent immunocytochemistry (FICC) allows multiple labeling approaches when enzyme-based techniques are difficult to combine, such as in double-labeling experiments targeting small-caliber axonal segments. Nevertheless, the conversion of FICC to a product visible at the electron microscopic (EM) level requires labor-intensive procedures, thus justifying the development of more user-friendly conversion methods. This study was initiated to simplify the conversion of FICC to EM by employing the unique properties of tyramide signal amplification (TSA), which allowed the simultaneous targeting of a fluorescent tag and biotin label to the same antigenic site. Briefly, one of two antigenic sites typically co-localized in damaged axonal segments was visualized by the application of a fluorescent secondary antibody, with the other tagged via a biotinylated antibody. Next, an ABC kit was used, followed by the simultaneous application of fluorophore-tyramide and biotin-tyramide. After temporary mounting for fluorescent digital photomicroscopy, sections were incubated in ABC and reacted with diaminobenzidine before EM analysis. Double-labeling fluorescent immunocytochemistry with TSA clearly delineated damaged axonal segments. In addition, these same axonal segments yielded high-quality EM images with discrete electron-dense reaction products, thereby providing a simple and reproducible means for following fluorescent analysis with EM.

show abstract

Immunocytochemistry of endothelial nitric oxide synthase in the rat brain: a light and electron microscopical study using the tyramide signal amplification technique

Cited by 85 publications

References 72 publications

Estrogen Increases Endothelial Nitric Oxide Synthase via Estrogen Receptors in Rat Cerebral Blood Vessels

Estrogen Increases Endothelial Nitric Oxide Synthase via Estrogen Receptors in Rat Cerebral Blood Vessels

Tonic and Phasic Nitric Oxide Signals in Hippocampal Long-Term Potentiation

Novel Application of Tyramide Signal Amplification (TSA): Ultrastructural Visualization of Double-labeled Immunofluorescent Axonal Profiles

Contact Info

Product

Resources

About