A monoclonal antibody stains myogenic cells in regenerating newt muscle

Development ◽  
1987 ◽  
Vol 101 (2) ◽  
pp. 267-277 ◽  
Author(s):  
K.J. Griffin ◽  
D.M. Fekete ◽  
B.M. Carlson
Keyword(s):  
Monoclonal Antibody ◽  
Muscle Regeneration ◽  
Tissue Repair ◽  
Limb Regeneration ◽  
Alternative Interpretation ◽  
Muscle Fibres ◽  
Muscle Repair ◽  
Contralateral Limb ◽  
Minced Muscle ◽  
Regeneration Blastema

Monoclonal antibodies have been used to study minced muscle regeneration in the adult newt, Notophthalmus viridescens. The contralateral limb was amputated and the immunostaining patterns in the regenerating blastema were compared with the minced tissue in sectioned material. Staining with a myofibre-specific antibody, called 12/101 (Kintner & Brockes, 1984), showed that myofibre degeneration was complete by 8–10 days after mincing, with myogenesis commencing 2 days later. Another monoclonal antibody, called 22/18, previously shown to label a subset of cells in the regeneration blastema of the newt (Kintner & Brockes, 1984, 1985), was found also to recognize a population of cells in regenerating minced muscle. At 6 days after mincing, the number of 22/18-positive (22/18+) cells was low but by days 12–16, during the period of myogenesis, their number had increased to become a major population within the minced tissue. A small number of the 22/18+ cells could be double labelled with 12/101 at this time. Prior to this, there was a phase in which 12/101 staining had disappeared from the mince. Cells immunoreactive with both antibodies after this phase confirm that at least some of the 22/18+ cells are myogenic. The number of 22/18+ cells decreased as muscle repair and maturation progressed. These results show that 22/18 is not specifically associated with blastemal cells but is a more general marker for regenerating systems in the newt. They further suggest an alternative interpretation of the double-labelled cells used by Kintner & Brockes (1984) as evidence for myofibre dedifferentiation in limb regeneration. Instead, we propose that such cells represent new myogenesis occurring by tissue repair of locally damaged muscle fibres.

Calcineurin-Aα activation enhances the structure and function of regenerating muscles after myotoxic injury

2007 ◽  
Vol 293 (2) ◽  
pp. R686-R694 ◽  
Author(s):  
Nicole Stupka ◽  
Jonathan D. Schertzer ◽  
Rhonda Bassel-Duby ◽  
Eric N. Olson ◽  
Gordon S. Lynch
Keyword(s):  
Muscle Regeneration ◽  
Muscle Repair ◽  
Contralateral Limb ◽  
Slow Fiber ◽  
Force Recovery ◽  
And Function ◽  
Muscle Contractile ◽  
Calcineurin Inhibition ◽  
Enhancer Factor

Calcineurin signaling is essential for successful muscle regeneration. Although calcineurin inhibition compromises muscle repair, it is not known whether calcineurin activation can enhance muscle repair after injury. Tibialis anterior (TA) muscles from adult wild-type (WT) and transgenic mice overexpressing the constitutively active calcineurin-Aα transgene under the control of the mitochondrial creatine kinase promoter (MCK-CnAα*) were injected with the myotoxic snake venom Notexin to destroy all muscle fibers. The TA muscle of the contralateral limb served as the uninjured control. Muscle structure was assessed at 5 and 9 days postinjury, and muscle function was tested in situ at 9 days postinjury. Calcineurin stimulation enhanced muscle regeneration and altered levels of myoregulatory factors (MRFs). Recovery of myofiber size and force-producing capacity was hastened in injured muscles of MCK-CnAα* mice compared with control. Myogenin levels were greater 5 days postinjury and myocyte enhancer factor 2a (MEF2a) expression was greater 9 days postinjury in muscles of MCK-CnAα* mice compared with WT mice. Higher MEF2a expression in regenerating muscles of MCK-CnAα* mice 9 days postinjury may be related to an increase of slow fiber genes. Calcineurin activation in uninjured and injured TA muscles slowed muscle contractile properties, reduced fatigability, and enhanced force recovery after 4 min of intermittent maximal stimulation. Therefore, calcineurin activation can confer structural and functional benefits to regenerating skeletal muscles, which may be mediated in part by differential expression of MRFs.

Cell cycle controls and the role of nerves and the regenerate epithelium in urodele forelimb regeneration: possible modifications of basic concepts

1987 ◽  
Vol 65 (8) ◽  
pp. 739-749 ◽  
Author(s):  
Roy A. Tassava ◽  
David J. Goldhamer ◽  
Bruce L. Tomlinson
Keyword(s):  
Cell Cycle ◽  
Monoclonal Antibody ◽  
Limb Regeneration ◽  
Blastema Formation ◽  
Basic Concepts ◽  
Regenerate Epithelium ◽  
Early Wound ◽  
Skin Epidermis ◽  
Wound Epithelium

Data from pulse and continuous labeling with [3H]thymidine and from studies with monoclonal antibody WE3 have led to the modification of existing models and established concepts pertinent to understanding limb regeneration. Not all cells of the adult newt blastema are randomly distributed and actively progressing through the cell cycle. Instead, many cells are in a position that we have designated transient quiescence (TQ) and are not actively cycling. We postulate that cells regularly leave the TQ population and enter the actively cycling population and vice versa. The size of the TQ population may be at least partly determined by the quantity of limb innervation. Larval Ambystoma may have only a small or nonexisting TQ, thus accounting for their rapid rate of regeneration. Examination of reactivity of monoclonal antibody WE3 suggests that the early wound epithelium, which is derived from skin epidermis, is later replaced by cells from skin glands concomitant with blastema formation. WE3 provides a useful tool to further investigate the regenerate epithelium.

Symmetrical vascularization patterns in normal and retinoic acid treated limb regenerates of the axolotl, Ambystoma mexicanum

1998 ◽  
Vol 76 (9) ◽  
pp. 1795-1796 ◽  
Author(s):  
Steven R Scadding ◽  
Andrew Burns
Keyword(s):  
Retinoic Acid ◽  
Pattern Formation ◽  
Vascular System ◽  
Limb Regeneration ◽  
Ambystoma Mexicanum ◽  
Limb Pattern ◽  
Regeneration Blastema

The purpose of this investigation was to determine whether there were any asymmetries in the vascularization of the limb-regeneration blastema in the axolotl, Ambystoma mexicanum, that might be related to pattern formation, and to determine if retinoic acid could modify the vascular patterns of the blastema. We used acrylic casts of the vascular system of the limbs to assess the pattern of vascularization. We observed a very regular symmetrical arrangement of capillaries in the limb-regeneration blastema that did not appear to be modified by doses of retinoic acid sufficient to modify the limb pattern.

scholarly journals Premature satellite cell activation before injury accelerates myogenesis and disrupts neuromuscular junction maturation in regenerating muscle

2020 ◽  
Vol 319 (1) ◽  
pp. C116-C128
Author(s):  
Nasibeh Daneshvar ◽  
Ryuichi Tatsumi ◽  
Jason Peeler ◽  
Judy E. Anderson
Keyword(s):  
Axon Guidance ◽  
Satellite Cell ◽  
Muscle Regeneration ◽  
Cell Activation ◽  
Neuromuscular Junctions ◽  
Muscle Repair ◽  
No Donor ◽  
Satellite Cell Activation ◽  
Models Of Injury ◽  
Terminal Schwann Cells

Satellite cell (SC) activation, mediated by nitric oxide (NO), is essential to myogenic repair, whereas myotube function requires innervation. Semaphorin (Sema) 3A, a neuro-chemorepellent, is thought to regulate axon guidance to neuromuscular junctions (NMJs) during myotube differentiation. We tested whether “premature” SC activation (SC activation before injury) by a NO donor (isosorbide dinitrate) would disrupt early myogenesis and/or NMJs. Adult muscle was examined during regeneration in two models of injury: myotoxic cardiotoxin (CTX) and traumatic crush (CR) ( n = 4–5/group). Premature SC activation was confirmed by increased DNA synthesis by SCs immediately in pretreated mice after CTX injury. Myotubes grew faster after CTX than after CR; growth was accelerated by pretreatment. NMJ maturation, classified by silver histochemistry (neurites) and acetylcholinesterase (AchE), and α-bungarotoxin staining (Ach receptors, AchRs) were delayed by pretreatment, consistent with a day 6 rise in the denervation marker γ-AchR. With pretreatment, S100B from terminal Schwann cells (TSCs) increased 10- to 20-fold at days 0 and 10 after CTX and doubled 6 days after CR. Premature SC activation disrupted motoneuritogenesis 8–10 days post-CTX, as pretreatment reduced colocalization of pre- and postsynaptic NMJ features and increased Sema3A-65. Premature SC activation before injury both accelerated myogenic repair and disrupted NMJ remodeling and maturation, possibly by reducing Sema3A neuro-repulsion and altering S100B. This interpretation extends the model of Sema3A-mediated motoneuritogenesis during muscle regeneration. Manipulating the timing and type of Sema3A by brief NO effects on SCs suggests an important role for TSCs and Sema3A-65 processing in axon guidance and NMJ restoration during muscle repair.

VIII.—Regeneration of the Legs of Decapod Crustacea from the Preformed Breaking Plane

1915 ◽  
Vol 35 ◽  
pp. 78-94 ◽  
Author(s):  
J. Herbert Paul
Keyword(s):  
Cell Proliferation ◽  
Hermit Crab ◽  
Functional Activity ◽  
Limb Regeneration ◽  
Single Layer ◽  
Shore Crab ◽  
Muscle Fibres ◽  
Rhythmic Movements ◽  
Small Vessels ◽  
Decapod Crustacea

Summary(1) Homarus vulgáris, Eupagurus bernhardus, and Carcinus mœnas all form limb-buds or papillæ in the process of limb regeneration. These are covered by a chitinous envelope, and the observations here recorded show that their outer form and size are adaptations to the requirements of the animal. That of the lobster is straight, that of the hermit crab curved, while the shore crab has a papilla which may be folded on itself three times inside the envelope.(2) Valvular action of the diaphragm at the breaking plane plays a greater part in the stopping of haemorrhage after self-amputation than clotting, and the dilatation of small vessels which pass beneath the epidermis detaches a layer of cells. This layer of epidermis proliferates from its free edges to form the new limb.(3) A new diaphragm is the first structure laid down, and differentiation takes place from the base outwards. Muscle arises at the growing tip from cells proliferated from the old epidermis (an ectodermal structure), and the nerve grows outwards from the torn end by cell proliferation.(4) Muscle-fibres are anatomically complete immediately before moulting. The fibrillæ are cross-striated and enclosed in a sarcolemma, but full functional activity does not come till several days after moulting, beginning with slow rhythmic movements. Sarcoplasm seems to be less plentiful than in the normal fibre.(5) When moulting occurs the papilla is at once expanded to several times its previous size by valvular action, and the epidermis, previously composed of several layers of cells, now thins to a single layer, as is seen in the normal limb.

scholarly journals Plasticity of Retrovirus-Labelled Myotubes in the Newt Limb Regeneration Blastema

2000 ◽  
Vol 218 (2) ◽  
pp. 125-136 ◽  
Author(s):  
Anoop Kumar ◽  
Cristiana P. Velloso ◽  
Yutaka Imokawa ◽  
Jeremy P. Brockes
Keyword(s):  
Limb Regeneration ◽  
Regeneration Blastema ◽  
Newt Limb
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