A new specific, sensitive and non-carcinogenic reagent for the demonstration of horseradish peroxidase

Hanker, Jacob S.; Yates, Peggy E.; Metz, Charles B.; Rustioni, Aldo

doi:10.1007/bf01003075

Cited by 940 publications

(208 citation statements)

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“…The dorsal column was then embedded in agar and cut longitudinally into sections 200 m thick using a vibratome. The HRP was visualized by exposure of the sections to a preincubation solution containing Hanker-Yates reagent (Hanker et al, 1977) (Sigma, Poole, Dorset, UK ; 0.5 mg /ml in 0.1 M cacodylate buffer) for 15 min, followed by a 15 min exposure to the same solution with the addition of 0.015% H 2 O 2 . If a stained axon was observed within the lesion on light microscopic examination, the lesion was excised, osmicated, dehydrated through ascending ethanols, passed through propylene oxide, and embedded in Epon 812 (Polysciences, Warrington, PA) using standard techniques.…”

Section: Methodsmentioning

confidence: 99%

Conduction in Segmentally Demyelinated Mammalian Central Axons

1997

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The prominent symptoms associated with central demyelinating diseases such as multiple sclerosis (MS) are primarily caused by conduction deficits in affected axons. The symptoms may go into remission, but the mechanisms underlying remissions are uncertain. One factor that could be important is the restoration of conduction to affected axons, but it is not known whether demyelinated central axons resemble their peripheral counterparts in being able to conduct in the absence of repair by remyelination. In the present study we have made intraaxonal recordings from central axons affected by a demyelinating lesion, and then the axons have been labeled ionophoretically to permit their subsequent identification. Ultrastructural examination of 23 labeled preparations has established that some segmentally demyelinated central axons can conduct, and that they can do so over continuous lengths of demyelination exceeding several internodes (2500 m). Such segmentally demyelinated central axons were found to conduct with the anticipated reduction in velocity and a refractory period of transmission (RPT) as much as 34 times the value obtained from the nondemyelinated portion of the same axon; the RPT was typically prolonged to 2-5 times the normal value. We conclude that some segmentally demyelinated central axons can conduct, and we propose that the restoration of conduction to such axons is likely to contribute to the remissions commonly observed in diseases such as MS. Key words: demyelination; multiple sclerosis; axon; conduction properties; glia; ionophoresisMultiple sclerosis (MS) is a disease of the C NS in which affected central axons often lose one or more internodes of their myelin sheath; the continuity of the axon through the lesion is frequently maintained, although degeneration becomes more prominent as the disease progresses (McDonald, 1994). The disease is associated with symptoms such as paralysis, blindness, and numbness, which can be explained by conduction block in the relevant pathways. Such symptoms may spontaneously go into remission; however, the mechanisms underlying such remissions are not well understood. It is now clear that some demyelinated lesions are partially repaired by remyelination and that this phenomenon can be quite common and extensive in early lesions (Prineas et al., 1993a). Remyelination is known to restore secure conduction to central (Smith et al., 1979(Smith et al., , 1981Blight and Young, 1989;Felts and Smith, 1992;Honmou et al., 1996) and peripheral (Saida et al., 1980;Smith and Hall, 1980;Sedal et al., 1983;Shrager, 1988) demyelinated axons, and it is reasonable to believe that conduction in remyelinated central axons will contribute to remissions; however, even where remyelination occurs in MS, it may be temporary (Prineas et al., 1993b), and persistently demyelinated lesions are a common feature of the disease. It is known that some of these persistently demyelinated lesions are clinically silent, i.e., they do not produce symptoms (Ghatak et al., 1974;Phadke and Best, 1983; f...

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Section: Methodsmentioning

confidence: 99%

Conduction in Segmentally Demyelinated Mammalian Central Axons

1997

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“…One section out of five was processed with p-phenylenediamine-pyrocathecol (Hanker et al, 1977) to visualize the injection sites, and mounted on slides.…”

Section: Methodsmentioning

confidence: 99%

Cervicothalamic tract terminals are enriched in glutamate-like immunoreactivity: an electron microscopic double-labeling study in the cat

Broman

Ottersen

1992

J. Neurosci.

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The distribution of glutamate-like immunoreactivity (Glu-LI) in the thalamic ventral posterolateral nucleus (VPL) of cats was studied with the EM immunogold technique in order to identify nerve terminal populations that may use glutamate as a neurotransmitter. The investigation was focused on cervicothalamic tract (CTT) terminals, which were labeled by WGA-HRP transported anterogradely from injection sites in the lateral cervical nucleus (LCN). The amount of Glu-LI in different profiles was evaluated quantitatively by counting the number of gold particles and then calculating the areal density of gold particles over different profile types. The highest density of gold particles was found over terminals with morphologic characteristics of terminals of cortical origin (RS terminals), a finding that further supports the glutamatergic nature of these terminals suggested by previous studies. Enrichment of Glu-LI was also found in CTT terminals and in non-peroxidase-labeled terminals with the same morphologic characteristics as CTT terminals (RL terminals). The labeling density over these terminals was about twice the average tissue density of gold particles. The labeling density over large VPL neuronal cell bodies was on average 127%, and that over vesicle-containing dendritic appendages and truncs (presynaptic dendrites) about 80%, of the average tissue density of gold particles. Immunogold labeling with antiserum against glutamine (Gln) indicated low levels of Gln-like immunoreactivity in CTT terminals and a high Glu:Gln ratio as compared to astrocytes and the average Glu:Gln ratio in the VPL. The present findings provide further support for a transmitter role of glutamate in terminals of ascending somatosensory afferents to the VPL, including the CTT. Taken together with previous findings of an enrichment of Glu-LI in terminals of the spinocervical tract (Broman et al., 1990), our results suggest that synaptic transmission in the spinocervicothalamic pathway is dependent on the release of glutamate both at the levels of the LCN and the VPL.

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“…One to two hours were allowed for the diffusion of HRP throughout the last cell that was injected; the ganglion was then fixed overnight in 1.25% glutaraldehyde and 0.5% paraformaldehyde. The following day, the HRP reaction product was developed by the method of Hanker et al (1977). Ganglia were subsequently dehydrated, cleared, and mounted whole on slides with DPX (BDH Chemicals, Ltd.).…”

Section: Methodsmentioning

confidence: 99%

Changes in the dendritic geometry of mouse superior cervical ganglion cells following postganglionic axotomy

Yawo

1987

J. Neurosci.

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The dendritic geometry of mouse superior cervical ganglion cells was studied over periods of up to 3 months after postganglionic axotomy. Intracellular injection of HRP showed that total dendritic length and complexity were reduced by 60–70%, on average, among cells whose postganglionic axons had been crushed 2 weeks before. Both parameters gradually recovered in parallel with ganglion cell reinnervation of the periphery. These results indicate that neuronal interactions with peripheral targets influence the configuration of ganglion cell dendrites throughout life. The implications of this conclusion are discussed.

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A new specific, sensitive and non-carcinogenic reagent for the demonstration of horseradish peroxidase

Cited by 940 publications

References 0 publications

Conduction in Segmentally Demyelinated Mammalian Central Axons

Conduction in Segmentally Demyelinated Mammalian Central Axons

Cervicothalamic tract terminals are enriched in glutamate-like immunoreactivity: an electron microscopic double-labeling study in the cat

Changes in the dendritic geometry of mouse superior cervical ganglion cells following postganglionic axotomy

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