Motorneuron pools innervating muscles in vitamin A-induced proximal-distal duplicate limbs in the axolotl

1985 ◽  
Vol 224 (1236) ◽  
pp. 341-354 ◽  
Keyword(s):  
Horseradish Peroxidase ◽  
Vitamin A ◽  
Nerve Regeneration ◽  
Retrograde Transport ◽  
Axonal Guidance ◽  
The Other ◽  
Limb Muscle ◽  
Hindlimb Muscle ◽  
Forelimb Muscles ◽  
Distal Limb Muscle

Serially duplicated limbs containing two sets of proximal muscles were created in axolotls by vitamin A treatment. The innervation of three replicated proximal muscles was studied by using retrograde transport of horseradish peroxidase. These were the forelimb muscles biceps (seven cases) and anconeus (five cases) and the hindlimb muscle puboischiotibialis (five cases). In two cases (both of anconeus) innervation was from a correct motorneuron pool. In the other 15 cases the innervation was from an incorrect, distal limb muscle, motorneuron pool. These results are interpreted as evidence against long range signals between nerve and muscle controlling specific nerve regeneration. However, the data are compatible with models of axonal guidance that use local pathway cues.

The pattern of innervation in serially duplicated axolotl limbs: further evidence for the existence of local pathway cues?

Development ◽  
1987 ◽  
Vol 100 (3) ◽  
pp. 479-487
Author(s):  
N. Stephens ◽  
N. Holder
Keyword(s):  
Spinal Cord ◽  
Horseradish Peroxidase ◽  
Local Environment ◽  
Retrograde Transport ◽  
Limb Muscle ◽  
Biceps Muscle ◽  
Motor Innervation ◽  
Nerve Sprouting ◽  
Correct Pattern ◽  
Motor Neurones

The innervation of the biceps muscle was examined in regenerated and vitamin A-induced serially duplicated axolotl forelimbs using retrograde transport of horseradish peroxidase. The regenerated biceps muscle becomes innervated by motor neurones in the same position in the spinal cord as the normal biceps motor pool. In previous experiments in which the innervation of a second copy of a proximal limb muscle was examined in serially duplicated limbs (Stephens, Holder & Maden, 1985), the duplicate muscle was found to become innervated by motor neurones that would normally have innervated distal muscles. In the present study, the innervation of the second copy of biceps was examined under conditions designed to encourage nerve sprouting from ‘correct’ biceps axons. Following either partial limb denervation or denervation coupled with removal of the proximal biceps, the second copy of the muscle was still innervated by inappropriate motor neurones, which again would normally innervate distal limb muscles. These results are interpreted as evidence for the necessity for an appropriate local environment for axonal growth to allow reformation of a correct pattern of motor innervation in the regenerated limb.

Comparative strengths and weaknesses of peroxidase and colloidal gold in pre- and post-embedding immunolabeling techniques

1987 ◽  
Vol 45 ◽  
pp. 1002-1005
Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Light Microscopy ◽  
Colloidal Gold ◽  
Light Microscope ◽  
Ultrastructural Localization ◽  
The Other ◽  
Quantitative Studies ◽  
Dense Suspension ◽  
Antigen Localization

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.

Ultrastructural Characteristics of Capillaries Entering and Leaving the Stria Vascularis

1986 ◽  
Vol 95 (3) ◽  
pp. 309-312 ◽  
Author(s):  
Kensuke Watanabe
Keyword(s):  
Horseradish Peroxidase ◽  
Basal Cell ◽  
Stria Vascularis ◽  
The Other ◽  
Spiral Ligament ◽  
Perivascular Spaces ◽  
Basal Cells ◽  
Vascular Flow ◽  
Cell Layers ◽  
Ultrastructural Characteristics

Capillaries entering and leaving the stria vascularis were surrounded by layers of basal cells and fibrocytes. The entering capillaries were surrounded by one or two thin basal cells, while the leaving capillaries were surrounded by four or five thicker and interdigitated basal cell layers. Moreover, the layers surrounding the leaving capillaries persisted further into the spiral ligament. Two kinds of filaments were observed in the basal cells, one thin and the other thick. Capillaries were observed to leak horseradish peroxidase before they entered and after they left the stria vascularis. Although the reaction product of horseradish peroxidase was observed in all perivascular spaces of leaving capillaries, very little or no reaction product was observed around some entering capillaries. It is speculated that the layers of basal cells and fibrocytes around entering and leaving capillaries control the vascular flow out of the stria vascularis, although the layers around leaving capillaries may be more contractile than those around entering capillaries.

Reformation of specific neuromuscular connections during axolotl limb regeneration: evidence that the first contacts are correct

Development ◽  
1988 ◽  
Vol 103 (2) ◽  
pp. 365-377
Author(s):  
S. Wilson ◽  
M. Jesani ◽  
N. Holder
Keyword(s):  
Spinal Cord ◽  
Horseradish Peroxidase ◽  
Limb Regeneration ◽  
Microscopic Analysis ◽  
Motor Neurone ◽  
Extensor Digitorum ◽  
Limb Muscle ◽  
Proximal Muscle ◽  
Electron Microscopic ◽  
Neuronal Tracing

Retrograde neuronal tracing with horseradish peroxidase was used to determine the position in the spinal cord of the motor neurone pools of a proximal (biceps) and a distal (extensor digitorum) limb muscle at various times during axolotl limb regeneration. It was found that from the earliest stages of muscle redifferentiation (as judged by light and electron microscopic analysis) the vast majority of axons innervating the regenerating muscles came from cells within the bounds of the normal motor neurone pool for each muscle. A few incorrect projections were noted in that the regenerating proximal muscle was sometimes innervated by some cells caudal to its normal motor neurone pool. The results are discussed in terms of mechanisms that may be operating in the regenerating limb to ensure that specific neuromuscular connections are made.

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