A sensitive low artifact TMB procedure for the demonstration of WGA-HRP in the CNS

Gibson, Alan R.; Hansma, D. I.; Houk, James C.; Robinson, Farrel R.

doi:10.1016/0006-8993(84)91423-9

Cited by 272 publications

(120 citation statements)

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“…Pentobarbitone, for example, is known to inhibit the transport of HRP (Mesulam and Mufson, 1980), and, in our hands, a survival time of less than 2 days fails to produce detectable retrograde labelling in the inferior olive after injection of WGA±HRP into the cerebellar cortex (Trott and Apps, 1993). By contrast, periods in excess of 5 days are considered likely to result in transneuronal labelling and an inaccurate estimation of the size of the eective injection site, as well as some decrease in labelling (Gibson et al, 1984;Mesulam, 1982). A survival period of between 3 and 5 days has therefore been used in studies of cortico-nuclear and olivo-cerebellar connections in the cat (Trott et al, 1998a(Trott et al, , 1998b, while 30±40 h is sucient for similar studies in rat (Apps, 1990).…”

Section: Wga-hrp Experiments and Electrophysiologymentioning

confidence: 59%

Current concepts in neuroanatomical tracing

Köbbert

Apps

Bechmann

et al. 2000

Progress in Neurobiology

380

336

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The development of new axonal tract tracing and cell labelling methods has revolutionised neurobiology in the last 30 years. The aim of this review is to consider some of the key methods of neuroanatomical tracing that are currently in use and have proved invaluable in charting the complex interconnections of the central nervous system. The review begins with a short overview of the most frequently used tracers, including enzymes, peptides, biocytin, latex beads, plant lectins and the everincreasing number of¯uorescent dyes. This is followed by a more detailed consideration of both well established and more recently introduced neuroanatomical tracing methods. Technical aspects of the application, uptake mechanisms, intracellular transport of tracers, and the problems of subsequent signal detection, are also discussed. The methods that are presented and discussed in detail include: (1) anterograde and retrograde neuroanatomical labelling with¯uorescent dyes in vivo, (2) labelling of post mortem tissue, (3) developmental studies, (4) transcellular tracing (phagocytosis-dependent staining of glial cells), (5) electrophysiological mapping combined with neuronal tract tracing, and (6) simultaneous detection of more than one axonal tracer. (7) Versatile protocols for three-colour labelling have been developed to study complex patterns of connections. It is envisaged that this review will be used to guide the readers in their selection of the most appropriate techniques to apply to their own particular area of interest. 7

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Section: Wga-hrp Experiments and Electrophysiologymentioning

confidence: 59%

Current concepts in neuroanatomical tracing

Köbbert

Apps

Bechmann

et al. 2000

Progress in Neurobiology

380

336

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“…Frozen 50 m coronal sections were arranged into four series and mounted on gelatin-coated slides. Two series were reacted for HRP (Gibson et al, 1984); one was stained with neutral red, and the other was coverslipped without staining. The two unreacted series were stained with cresyl violet or processed for parvalbumin (Carmichael and Price, 1994).…”

Section: Methodsmentioning

confidence: 99%

Gustatory Neural Responses in the Medial Orbitofrontal Cortex of the Old World Monkey

Pritchard¹,

Edwards²,

Smith³

et al. 2005

J. Neurosci.

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The primary taste cortex has widespread and occasionally dense projections to the orbitofrontal cortex (OFC) in the macaque. Nonetheless, electrophysiological studies have revealed that only 2-8% of the cells in the OFC are activated by taste stimuli on the tongue. We describe an area centered in Brodmann's area 13m of the medial OFC (mOFC) where taste neurons are more concentrated. It consists of a 12 mm 2 core, where gustatory neurons constituted 20% of the population, and a 1 mm perimeter in which 8% of the cells responded to taste. Data were collected from three awake cynomolgus monkeys (Macaca fascicularis) prepared for chronic recording. Single neurons were isolated with epoxylite-coated tungsten microelectrodes and tested for responsiveness to 1.0 M glucose, 0.3 M NaCl, 0.03 M HCl, and 0.001 M QHCl. These stimuli elicited responses that were 96% excitatory and ranged from 5.2 to 5.9 spikes/s. Cells were broadly tuned (H ϭ 0.79), similar to those in the anterior insula (H ϭ 0.70), and decidedly unlike the narrowly tuned taste neurons in the caudolateral OFC (clOFC; H ϭ 0.39). Whereas 82% of the taste cells in the clOFC respond to glucose, in the mOFC, HCl-responsive (56%), glucose-responsive (50%), NaCl-responsive (43%), and QHCl-responsive (40%) cells were almost evenly represented. The mOFC taste area appears to comprise a major gustatory relay that lies anatomically and functionally between the anterior insula and the clOFC.

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“…Alternate sections were processed for visualization of transported label using a modified tetramethylbenzidine (TMB) procedure of Gibson et al (1984). The remaining sections were used for neuroanatomical identification and were processed for cytochrome oxidase (CO) using the methods of Wong-Riley and Carroll (1984), or for myelin using the gold chloride technique of Schmued (1990).…”

Section: Histological Processingmentioning

confidence: 99%

Central visual system of the naked mole‐rat (Heterocephalus glaber)

Crish

Catania

2006

Anat. Rec.

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Naked mole-rats are fossorial rodents native to eastern Africa that spend their lives in extensive subterranean burrows where visual cues are poor. Not surprisingly, they have a degenerated eye and optic nerve, suggesting they have poor visual abilities. However, little is known about their central visual system. To investigate the organization of their central visual system, we injected a neuronal tracer into the eyes of naked mole-rats and mice to compare the neural structures mediating vision. We found that the superior colliculus and lateral geniculate nucleus were severely atrophied in the naked mole-rat. The olivary pretectal nucleus was reduced but still retained its characteristic morphology, possibly indicating a role in light detection. In addition, the suprachiasmatic nucleus is well innervated and resembles the same structure in other rodents. The naked mole-rat appears to have selectively lost structures that mediate form vision while retaining structures needed for minimal entrainment of circadian rhythms. Similar results have been reported for other mole-rat species. Taken together, these data suggest that light detection may still play an important role in the lives of these "blind" animals: most likely for circadian entrainment or setting seasonal rhythms. © 2006 Wiley-Liss, Inc.Key words: brain; evolution; vision; superior colliculus; eye; blind A well-developed visual system is metabolically costly, so one would expect a reduction in the visual systems of subterranean rodents where light levels are minimal. However, animals with occasional access to the surface can make use of brief light exposure for setting circadian and seasonal rhythms. Studies of both Ansell's mole-rat (Cryptomys anselli) and the blind mole-rat (Spalax ehrenbergi) have revealed severe atrophy of many of the structures involved in form vision, yet some of the structures involved in other types of light detection appear unchanged. This has led to the suggestion that light cues are still used to mediate aspects of behavior in at least some fossorial species (Cooper et al., 1993;Nemec et al., 2004).Naked mole-rats ( Fig. 1) are fossorial rodents from eastern Africa in the same family (Bathyergidae) as Ansell's mole-rat. Naked mole-rats live underground in large eusocial colonies with division of labor and separate castes that have different social, morphological, and behavioral characteristics. Only a few colony members are reproductively active, which results in a high level of genetic similarity in the colony due to inbreeding (Jarvis, 1981).In addition to social and physiological adaptations to life underground, mole-rats exhibit several sensory modifications. Sound attenuates rapidly underground, and presumably because of this environmental constraint, molerats have degraded hearing with poor sound localization abilities (Heffner and Heffner, 1993;Brittan-Powell et al., 2001). However, naked mole-rats are somatosensory specialists with a novel array of somatic vibrissae for guiding orienting behavior (Crish et al...

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A sensitive low artifact TMB procedure for the demonstration of WGA-HRP in the CNS

Cited by 272 publications

References 16 publications

Current concepts in neuroanatomical tracing

Current concepts in neuroanatomical tracing

Gustatory Neural Responses in the Medial Orbitofrontal Cortex of the Old World Monkey

Central visual system of the naked mole‐rat (Heterocephalus glaber)

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