scholarly journals Radioimmunocytochemistry using a tritiated goat anti-rabbit second antibody.

1984 ◽  
Vol 32 (7) ◽  
pp. 778-782 ◽  
Author(s):  
E J Glazer ◽  
J Ramachandran ◽  
A I Basbaum
Keyword(s):  
Substance P ◽  
Epoxy Resin ◽  
Immunoglobulin G ◽  
Spinal Dorsal Horn ◽  
Ultrastructural Level ◽  
Microscopic Level ◽  
Light Microscopic Level ◽  
Spinal Dorsal ◽  
Double Label ◽  
Polyclonal Antisera

Affinity-purified goat anti-rabbit immunoglobulin G (GAR) was conjugated with (3H)-propionyl succinimidate and used to localize substance P (SP), enkephalin (ENK), and serotonin immunoreactive sites in the spinal dorsal horn and medulla of the rat and cat. Autoradiographic localization was demonstrated on paraffin, frozen, Vibratome, and 2 micron plastic sections. The latter were obtained from radiolabeled Vibratome sections that were embedded in epoxy resin. The distribution of SP, ENK, and serotonin demonstrated by radioimmunocytochemistry was comparable to that observed on semiadjacent sections using peroxidase-antiperoxidase (PAP) immunocytochemistry. The autoradiograms, however, were generated using primary antibody concentrations up to five times more dilute than concentrations used for the PAP procedure. Indirect radioimmunocytochemistry using a (3H) anti-immunoglobulin G second antibody can be used to localize a variety of monoclonal and polyclonal antisera. It is quantifiable at the light microscopic level and can be potentially used with peroxidase histochemistry to double label immunoreactive structures at the ultrastructural level.

scholarly journals Biotinylphallotoxins: preparation and use as actin probes.

1989 ◽  
Vol 37 (7) ◽  
pp. 1035-1045 ◽  
Author(s):  
H Faulstich ◽  
S Zobeley ◽  
U Bentrup ◽  
B M Jockusch
Keyword(s):  
Actin Filaments ◽  
Ultrastructural Level ◽  
Microscopic Level ◽  
Electron Microscopic Level ◽  
Electron Microscopic ◽  
Permeabilized Cells ◽  
High Affinity ◽  
Double Label ◽  
The One

We describe the synthesis of four phalloidin derivatives conjugated with biotin. An aminomethyldithiolane derivative of ketophalloidin was used as a reactive starter compound, and biotin residues were coupled to this molecule either directly, separated by spacer chains comprised of one or two glycyl residues, or of a 12-atom long chain constructed from succinic acid and hexamethylendiamine. Although all products still displayed a high affinity for F-actin, as seen in competition experiments with [3H]-demethylphalloidin, only the one with the longest spacer (BHPP) showed specific and high-affinity decoration of actin filaments in permeabilized cells, in conjunction with FITC-coupled avidin and fluorescence microscopy. Combined with gold-streptavidin, BHPP decorated the actin filament system at the light and electron microscopic level faithfully and with satisfactory density. Actin filaments polymerized in vitro from purified protein were not as densely labeled as had been expected. However, in all these experiments the new phalloidin probe, when combined with avidin or streptavidin, yielded clear and highly specific labeling of F-actin. Therefore, this system is useful to identify and localize actin unambiguously in microfilaments, independent of actin antibodies, and should facilitate double-label experiments on cytoskeletal components at the ultrastructural level.

Effects of substance P analogues on spinal dorsal horn neurons

Peptides ◽  
1988 ◽  
Vol 9 (3) ◽  
pp. 651-660 ◽  
Author(s):  
M. Randic ◽  
S. Jeftinija ◽  
L. Urban ◽  
C. Raspantini ◽  
K. Folkers
Keyword(s):  
Substance P ◽  
Dorsal Horn ◽  
Spinal Dorsal Horn ◽  
Dorsal Horn Neurons ◽  
Spinal Dorsal ◽  
Spinal Dorsal Horn Neurons ◽  
Substance P Analogues

Antagonism of substance P induced facilitation of central neuronal activity by 2,4-dinitrophenol and depressant agents

1978 ◽  
Vol 56 (3) ◽  
pp. 527-530 ◽  
Author(s):  
Bhagavatula R. Sastry
Keyword(s):  
Nitrous Oxide ◽  
Substance P ◽  
Dorsal Horn ◽  
Neuronal Activity ◽  
Spinal Dorsal Horn ◽  
Cortical Cells ◽  
Dorsal Horn Neurons ◽  
Spinal Dorsal ◽  
Central Neurons ◽  
Spinal Dorsal Horn Neurons

2,4-Dinitrophenol, pentobarbital, thiopental, methoxyflurane, and halothane more often antagonized the facilitatory effects of substance P than of acetylcholine on the activity of spinal dorsal horn neurons (of unanaesthetized decerebrated and spinalized cats) as well as interpeduncular and cerebral cortical cells (of either rats anaesthetized with methoxyflurane, nitrous oxide, and oxygen or 'cerveau isolé' rats). The results of the studies indicate that the excitatory effects of substance P on central neurons are extremely sensitive to anaesthesia.

scholarly journals Regulation by opioidergic interneurons of substance P release in rat spinal dorsal horn slices

1987 ◽  
Vol 43 ◽  
pp. 191
Author(s):  
Shigeki Kawabata ◽  
Ryoya Oku ◽  
Kazuhiro Okochi ◽  
Masamichi Satoh ◽  
Hiroshi Takagi
Keyword(s):  
Substance P ◽  
Dorsal Horn ◽  
Spinal Dorsal Horn ◽  
Spinal Dorsal ◽  
P Release ◽  
Substance P Release
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