Selective illumination of single photoreceptors in the house fly retina: local membrane turnover and uptake of extracellular horseradish peroxidase (HRP) and Lucifer Yellow

1989 ◽  
Vol 257 (3) ◽  
pp. 565-576 ◽  
Author(s):  
Hansj�rg Wunderer ◽  
Serge Picaud ◽  
Nicolas Franceschini
Keyword(s):  
Horseradish Peroxidase ◽  
Lucifer Yellow ◽  
House Fly ◽  
Membrane Turnover

scholarly journals Phorbol esters and horseradish peroxidase stimulate pinocytosis and redirect the flow of pinocytosed fluid in macrophages.

1985 ◽  
Vol 100 (3) ◽  
pp. 851-859 ◽  
Author(s):  
J A Swanson ◽  
B D Yirinec ◽  
S C Silverstein
Keyword(s):  
Steady State ◽  
Horseradish Peroxidase ◽  
Phorbol Esters ◽  
Lucifer Yellow ◽  
Fluid Phase ◽  
Peritoneal Macrophages ◽  
Tumor Promoter ◽  
Steady State Rate ◽  
Reduced Rate ◽  
Rate Of Accumulation

Lucifer Yellow CH (LY) is an excellent probe for fluid-phase pinocytosis. It accumulates within the macrophage vacuolar system, is not degraded, and is not toxic at concentrations of 6.0 mg/ml. Its uptake is inhibited at 0 degree C. Thioglycollate-elicited mouse peritoneal macrophages were found to exhibit curvilinear uptake kinetics of LY. Upon addition of LY to the medium, there was a brief period of very rapid cellular accumulation of the dye (1,400 ng of LY/mg protein per h at 1 mg/ml LY). This rate of accumulation most closely approximates the rate of fluid influx by pinocytosis. Within 60 min, the rate of LY accumulation slowed to a steady-state rate of 250 ng/mg protein per h which then continued for up to 18 h. Pulse-chase experiments revealed that the reduced rate of accumulation under steady-state conditions was due to efflux of LY. Only 20% of LY taken into the cells was retained; the remainder was released back into the medium. Efflux has two components, rapid and slow; each can be characterized kinetically as a first-order reaction. The kinetics are similar to those described by Besterman et al. (Besterman, J. M., J. A. Airhart, R. C. Woodworth, and R. B. Low, 1981, J. Cell Biol. 91:716-727) who interpret fluid-phase pinocytosis as involving at least two compartments, one small, rapidly turning over compartment and another apparently larger one which fills and empties slowly. To search for processes that control intracellular fluid traffic, we studied pinocytosis after treatment of macrophages with horseradish peroxidase (HRP) or with the tumor promoter phorbol myristate acetate (PMA). HRP, often used as a marker for fluid-phase pinocytosis, was observed to stimulate the rate of LY accumulation in macrophages. PMA caused an immediate four- to sevenfold increase in the rate of LY accumulation. Both HRP and PMA increased LY accumulation by stimulating influx and reducing the percentage of internalized fluid that is rapidly recycled. A greater proportion of endocytosed fluid passes into the slowly emptying compartment (presumed lysosomes). These experiments demonstrate that because of the considerable efflux by cells, measurement of marker accumulation inaccurately estimates the rate of fluid pinocytosis. Moreover, pinocytic flow of water and solutes through cytoplasm is subject to regulation at points beyond the formation of pinosomes.

Mapping of neuronal contacts with intracellular injection of horseradish peroxidase and Lucifer yellow in combination

1981 ◽  
Vol 217 (1) ◽  
pp. 143-149 ◽  
Author(s):  
E.R. Macagno ◽  
K.J. Muller ◽  
W.B. Kristan ◽  
S.A. Deriemer ◽  
R. Stewart ◽  
...  
Keyword(s):  
Horseradish Peroxidase ◽  
Lucifer Yellow ◽  
Intracellular Injection

scholarly journals Combination of horseradish peroxidase and lucifer yellow staining for selective labeling of neurons at the electron microscopic level.

1991 ◽  
Vol 39 (11) ◽  
pp. 1579-1583 ◽  
Author(s):  
L Nonnotte ◽  
A Buisson ◽  
F Nagy ◽  
M Moulins
Keyword(s):  
Horseradish Peroxidase ◽  
Lucifer Yellow ◽  
Electron Microscopic Level ◽  
Reaction Products ◽  
Stomatogastric Nervous System ◽  
Electron Microscopic ◽  
Double Labeling ◽  
Selective Labeling ◽  
Experimental Neuroanatomy ◽  
Labeling Method

We have developed a new double labeling method for electron microscopy to characterize selectively two physiologically identified neurons on the same preparation. The stomatogastric nervous system of crustaceans was used to test the distinguishing staining characteristics of the two labels. Neurons were labeled on one side with horseradish peroxidase (HRP) and on the other side with Lucifer yellow (LY). After blue light irradiation of the tissue in the presence of diaminobendizine, the two labeled neurons could be easily observed and discriminated on the same section by the two different reaction products. This simple technique of double labeling is useful in experimental neuroanatomy for the detailed study of synaptic relationships.

Neuronal mapping: a photooxidation reaction makes Lucifer yellow useful for electron microscopy

Science ◽  
1982 ◽  
Vol 217 (4563) ◽  
pp. 953-955 ◽  
Author(s):  
AR Maranto
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Blue Light ◽  
Lucifer Yellow ◽  
Photooxidation Reaction

Irradiation Lucifer yellow-filled neurons with intense blue light in the presence of 3,3'-diaminobenzidine produces an electron-opaque osmiophilic polymer within the injected cells. This technique is valuable when cobalt or horseradish peroxidase injections are difficult or when a second intracellular marker is needed to demonstrate neuronal contacts.

Colonic mechanosensory afferent input to neurons in the mouse superior mesenteric ganglion

1997 ◽  
Vol 272 (2) ◽  
pp. G357-G366 ◽  
Author(s):  
S. M. Miller ◽  
J. H. Szurszewski
Keyword(s):  
Horseradish Peroxidase ◽  
Synaptic Input ◽  
Lucifer Yellow ◽  
Distal Colon ◽  
Afferent Input ◽  
Evoked Responses ◽  
Mesenteric Ganglion ◽  
Colonic Distension ◽  
Synaptic Responses

Electrical activity and synaptic responses were recorded intracellularly in 415 neurons of the mouse superior mesenteric ganglion (SMG) attached to a segment of distal colon in vitro. Eighty-seven percent of neurons tested received ongoing nicotinic cholinergic fast excitatory postsynaptic potentials (fEPSPs). Colonic distension caused an initial transient followed by a sustained, slowly adapting increase in fEPSP activity. Application of hexamethonium only to the colon reduced, but did not completely abolish, distension-evoked responses, suggesting direct projection of some distension-sensitive fibers. Ongoing fEPSPs were abolished when nerve trunks connecting the SMG to the colon were transected or blocked with tetrodotoxin applied to the colon. Intracellular labeling with horseradish peroxidase or lucifer yellow revealed that about 90% of neurons receiving colonic synaptic input had a caudally projecting axon; about 60% that did not receive colonic input had a rostrally projecting axon. The latter neurons were found only in the cephalad ganglion region. These results show that mouse SMG neurons receive colonic mechanosensory afferent synaptic input and thus may participate in sympathetic intestinal reflexes.

scholarly journals The Organization and Role During Locomotion of the Proximal Musculature of the Cricket Foreleg: I. Anatomy and Innervation

1986 ◽  
Vol 123 (1) ◽  
pp. 255-283
Author(s):  
GILLES LAURENT ◽  
DANIEL RICHARD
Keyword(s):  
Horseradish Peroxidase ◽  
Lucifer Yellow ◽  
Gryllus Bimaculatus ◽  
Motor Neurones ◽  
Dorsal Unpaired Median ◽  
Anterior Posterior

Reprint requests should be sent to D. Richard at this address. The structure of the proximal segments of the cricket (Gryllus bimaculatus) foreleg, together with the associated musculature and its innervation are described. The morphology of 50 motor neurones involved in the control of this musculature has been revealed using backfilling techniques with cobalt, horseradish peroxidase and Lucifer Yellow. The ‘ball and socket’ pleurocoxal joint is moved by three sets of anatomical antagonists (promotor-remotor, abductor-adductor, anterior-posterior rotator muscles) inserted on each side of the three axes of rotation. The axial coxotrochanteral joint is moved by the intrinsic levator and the depressor muscles; these depressors are composed of an intrinsic (coxotrochanteral) and a ‘double’ (pleurotrochanteral) subgroup. The double depressors, and all the muscles inserting on the trochantin (promotors) or the anterior coxal rim (adductor, abductors, anterior rotators) are supplied by at least eighteen neurones, whose axons run in nerve 3. The muscles that insert on the posterior coxal rim (remotors, posterior rotators) are innervated by at least twelve similar neurones whose axons run in nerve 4. The intrinsic coxal muscles are supplied by branches of nerve 5 (ten motor neurones to the levators, two to the depressors). Three presumably common inhibitors, and one Dorsal Unpaired Median (DUM) neurone have also been found.

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