Tracing axons and axon collaterals of spinal neurons using intracellular injection of horseradish peroxidase

Science ◽  
1976 ◽  
Vol 191 (4224) ◽  
pp. 312-313 ◽  
Author(s):  
P. Snow ◽  
P. Rose ◽  
A. Brown
Keyword(s):  
Horseradish Peroxidase ◽  
Spinal Neurons ◽  
Intracellular Injection ◽  
Axon Collaterals

The shapes of sensory and motor neurones and the distribution of their synapses in ganglia of the leech: a study using intracellular injection of horseradish peroxidase

1976 ◽  
Vol 194 (1117) ◽  
pp. 481-499 ◽  
Keyword(s):  
Horseradish Peroxidase ◽  
Sensory Cells ◽  
Main Process ◽  
Mechanical Stimuli ◽  
Sensory Neurones ◽  
Intracellular Injection ◽  
Blue Stain ◽  
Motor Neurones ◽  
Segmental Ganglion ◽  
Pass Through

Three types of sensory neurones and two kinds of motor neurones in the segmental ganglion of the leech were examined with the light and electron microscope after intracellular injection of horseradish peroxidase (HRP) for a histological marker. The aim was to develop a method for identifying the synapses of specific cells in the ganglion’s complex neuropil and to form a picture of their distribution and structure. Reaction of HRP with different benzidine derivatives produces opaque and electron dense deposits. For light microscopy a blue stain is formed that makes processes visible in whole mounts millimeters away from the injection site at the soma. The reaction product for electron microscopy is distributed throughout the cytoplasm, yet ultrastructural details are preserved. The sensory neurones that respond specifically to touch or pressure or noxious mechanical stimuli to the skin share in their branching pattern a number of common features. A single process arising from each cell body forms large primary branches that pass through the neuropil and leave the ganglion by the ipsilateral connectives and roots. Within the neuropil these branches give rise to numerous smaller secondary processes. In contrast, the annulus erector and large longitudinal motoneurones send their main process across the ganglion to bifurcate and enter the contralateral roots. Secondary processes of the motoneurones are highly branched and more numerous than those of the sensory cells. Each type of sensory and motor cell is distinguished by the shape, length and distribution of its secondary processes. Secondary processes of sensory neurones exhibit numerous swellings and irregularly shaped fingers. Electron micrographs show that the sensory neurones make synapses at these specializations, each of which contacts several postsynaptic processes. The sensory neurones receive inputs at the same fingers and swellings, an arrangement suggesting that regions within a cell’s arborization may function semi-autonomously. The main process and large branches of the two motor neurones are studded with spines a few micrometres long and a fraction of a micrometre in diameter. Vesicle-containing varicosities from other cells make synaptic contact primarily with the spines, which themselves have few vesicles. These two motor neurones are largely, if not entirely, postsynaptic to other neurones within the leech nervous system.

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
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