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PubMed2003

Expression of complement 3 and complement 5 in newt limb and lens regeneration.

Some urodele amphibians possess the capacity to regenerate their body parts, including the limbs and the lens of the eye. The molecular pathway(s) involved in urodele regeneration are largely unknown. We have previously suggested that complement may participate in limb regeneration in axolotls. To further define its role in the regenerative process, we have examined the pattern of distribution and spatiotemporal expression of two key components, C3 and C5, during limb and lens regeneration in the newt Notophthalmus viridescens. First, we have cloned newt cDNAs encoding C3 and C5 and have generated Abs specifically recognizing these molecules. Using these newt-specific probes, we have found by in situ hybridization and immunohistochemical analysis that these molecules are expressed during both limb and lens regeneration, but not in the normal limb and lens. The C3 and C5 proteins were expressed in a complementary fashion during limb regeneration, with C3 being expressed mainly in the blastema and C5 exclusively in the wound epithelium. Similarly, during the process of lens regeneration, C3 was detected in the iris and cornea, while C5 was present in the regenerating lens vesicle as well as the cornea. The distinct expression profile of complement proteins in regenerative tissues of the urodele lens and limb supports a nonimmunologic function of complement in tissue regeneration and constitutes the first systematic effort to dissect its involvement in regenerative processes of lower vertebrate species.

Amino Acid Sequence↗
PubMed1999

[Retinal regeneration after dissection of the optic nerve in newts exposed on board the Bion-11 biosatellite].

The work brought up initial information on the impacts of space flight (SF) on regeneration of nerve tissues in vertebrata. Summarized are data of analysis of the retinal regeneration following section of the ocular nerve and blood vessels in space-flown adult newts (Pleurodeles waltlii). Two weeks in SF were found not to impede the regeneration of retina as its growth was fully dependent on the same cell sources as in the condition of 1 g. In the newts which had been operated 2 wk prior to launch, recovery of retina in SF proceeded more intensively (phases V-VI) compared with the synchronous controls (phase IV). According to the morphometric analysis, differentiation of regenerates' layers in the space animals was also a more rapid process. The proliferative activity of cells in regenerates estimated with the 3H-timidine radioautography turned to be higher, too: the labeled nuclei index in early non-differentiated regenerates was in 1.2 to 1.5 times higher than in the control. Immunohistochemical array with the help of GFAP antibodies performed at the late phases of regeneration revealed an activating effect of SF on the Muller glia cells. These findings indicate that microgravity can stimulate general retinal regeneration and activate regenerate cells, specifically those involved in morphogenesis.

Animals↗
PubMed2000

Brief electrical stimulation promotes the speed and accuracy of motor axonal regeneration.

Functional recovery is often poor despite the capacity for axonal regeneration in the peripheral nervous system and advances in microsurgical technique. Regeneration of axons in mixed nerve into inappropriate pathways is a major contributing factor to this failure. In this study, we use the rat femoral nerve model of transection and surgical repair to evaluate (1) the effect of nerve transection on the speed of regeneration and the generation of motor-sensory specificity, (2) the efficacy of electrical stimulation in accelerating axonal regeneration and promoting the reinnervation of appropriate muscle pathways by femoral motor nerves, and (3) the mechanism of action of electrical stimulation. Using the retrograde neurotracers fluorogold and fluororuby to backlabel motoneurons that regenerate axons into muscle and cutaneous pathways, we found the following. (1) There is a very protracted period (10 weeks) of axonal outgrowth that adds substantially to the delay in axonal regeneration (staggered regeneration). This process of staggered regeneration is associated with preferential motor reinnervation (PMR). (2) One hour to 2 weeks of 20 Hz continuous electrical stimulation of the parent axons proximal to the repair site dramatically reduces this period (to 3 weeks) and accelerates PMR. (3) The positive effect of short-term electrical stimulation is mediated via the cell body, implicating an enhanced growth program. The effectiveness of such a short-period low-frequency electrical stimulation suggests a new therapeutic approach to accelerate nerve regeneration after injury and, in turn, improve functional recovery.

Animals↗
PubMed2002

[Influence of thinning on regeneration in a coastal pinus thunbergii forest].

A coastal forest planted nearby the sea can provide many shelter benefits for the coastal regions. It is ideally if the continuity of the shelter benefits could be preserved through reasonable management. Thinning and regeneration as the most important management techniques for plantations can help the continuity of the shelter benefits of the coastal forest. However, because of the peculiarities of coastal forest, i.e., the coastal plantation nearby the sea is vulnerable to disturbances (thinning as one kind of disturbance), the study on thinning and regeneration within a coastal forest is poorly understood. The purpose of this paper is to give a primary understanding in natural regeneration for the coastal Pinus thunbergii forest with different thinning rates after four growing seasons since thinning. The experiment was carried out at the middle of the shoreline along the Japan Sea, and the investigated sites consisted of four thinning treatments (control, 0% thinned, 20% thinned, 30% thinned and 50% thinned) in a coastal P. thunbergii forest. After thinning, the regenerated seedlings, soil water content, light condition (canopy openness or canopy density), wind regime, and litter depth and quantity were investigated for four growing seasons. The relationships between the regenerated seedlings and light condition, litter, wind profile and soil water content were examined. The results showed that thinning could improve the light condition on the forest floor, increase the exchange of airflow (wind speed) in the coastal forest stand, and ameliorate the water content of the forest soil. These factors accelerated the decomposition of litters, and provided necessary conditions for natural regeneration. The results of regeneration observation indicated that the most intensively thinned treatment (50% thinned with density of about 1500 stems.hm-2) could provide a better condition for regeneration during the four growing seasons. The density and growth of seedling (greater than 1 year) increased significantly with increasing thinning intensities, and the establishment of seedlings was obviously succeeded in the most intensively thinned treatment, but failed in less thinned treatments and understory. The thinned intensity of 50% did not induce wind damage to the coastal forest in the four years after thinning, and did not cause the loss of shelter functions of the coastal forest such as sand blocking, wind breaking and salt preventing etc.. On the contrary, it could provide the suitable conditions for natural regeneration of the pine coastal forest, or for the immigration of other species. Therefore, thinning as the silviculturally created openness is very important for the establishment of seedlings in the coastal forests, which provides a mechanism for the coastal forest from even aged stands dominated by P. thunbergii to stands containing multiple size classes and canopy layers.

China↗
PubMedDate not supplied

[Age factor in eye regeneration of the gastropod mollusk Achatina fulica].

The dependence of the ability to regenerate the eye on the age of experimental animals was studied in the snail Achatina fulica. The degree of regeneration was estimated by light-microscopic and electrophysiological methods and by analyzing the motor response to visual stimuli. In older age groups, the number of regenerated eye-bearing tentacles decreased, whereas the period of regeneration increased. The regenerated eyes of the snails operated at the age of more than two months remained smaller than normal eyes even after six months. Regeneration of the distal part of the optic nerve was observed, and the regenerated eyes recovered the ability to respond to stimulation by light. In the electroretinogram, the responses of the regenerated eye, compared to the control, were characterised by a lower amplitude and longer repolarization and refractory periods. Manifestations of the motor response to visual stimuli in the young snails with regenerating eyes could be regarded as evidence for the recovery of connection between the organ of sight and the central ganglia.

Age Factors↗
PubMed2003

[Transient compensatory hypertrophy and limited regeneration of the surgically removed rat ventral prostate].

OBJECTIVES: To confirm whether regeneration of prostate lobe indeed takes place on surgical lobectomy and if so, to what extent. Other issues studied are 1. whether the lobe regenerated is similar morphologically to that developing normally from neonatal origin to adulthood, and 2. the consequences of partial lobectomy on the contralateral lobe and the influence of sex steroid hormones on the regeneration process. METHODS: The effect of surgical removal of one of the ventral prostate lobes on the size of the contralateral lobe has been studied at various time intervals after lobectomy. RESULTS: The surgically extirpated ventral prostate lobe in rats regenerates attaining plateau size at 8-16 weeks post lobectomy. The regenerated lobe, however, remains significantly smaller than the original size. In early phase of post lobectomy (at 2 weeks) the contralateral lobe was significantly hypertrophied. It reverts to normal size on regeneration of the extirpated lobe with time. Orchiectomy carried out at the time of lobectomy caused a drastic reduction in the size of the remaining lobe, which was prevented by exogenous treatment with androgens. In animals receiving treatment with estrogens, the remaining lobe was partially but not fully atrophied. However, estrogens did not support the regeneration of the surgically removed lobe, which requires androgens. CONCLUSIONS: These studies demonstrate that surgical removal of one of the ventral prostate lobe leads to a process of regeneration. However, the regenerated lobe does not attain the normal size.

Animals↗
PubMed2005

Arginin-vasopressin regulates proliferative activity of the regenerating rat adrenal cortex.

Enucleation-induced adrenal regeneration is a classic model to investigate adrenocortical proliferation in vivo, which is dependent not only on pituitary ACTH release, but also on various other neural and endocrine signals. Arginin-vasopressin (AVP), mainly acting via V1 receptors, regulates hypothalamic-hypophyseal-adrenal axis function, acting on both its central and peripheral branches. Here, we studied whether endogenous AVP system modulates rat adrenal regeneration. Reverse transcription-polymerase chain reaction (PCR) detected only the mRNAs of V1a and V1b receptors in normal and regenerating adrenals. The expression was very low, and semi-quantitative conventional and real-time PCR showed that it was down-regulated in regenerating adrenals in relation to the time elapsed from enucleation. AVP (three subcutaneous injections 28, 16 and 4 h before sacrifice) raised metaphase index at day 5, but not at day 8 of regeneration. Unexpectedly, both V1-receptor and V2-receptor antagonists increased metaphase index at days 5 and 8 of regeneration. Neither AVP nor AVP-receptor antagonists affected plasma levels of corticosterone in rats bearing regenerating adrenals. It is concluded that AVP, acting via V1 receptors located in adrenals, exerts a stimulating effects on adrenal regeneration. Due to the down-regulation of V1-receptor expression in regenerating adrenals, this effect is very weak and is easily overcome by a tonic inhibitory action of endogenous AVP systems probably involving extra-adrenal indirect mechanisms.

Adrenal Cortex↗
PubMed2005

[Reparative regeneration of nerve fibers after sympathectomy: an experimental study].

The results of sympathectomies depend on the degree of regeneration of the sympathetic nerve trunk: the quicker the onset of regeneration the less remarkable the effect of operation. The aim of our experimental study WPS to examine the rate of regeneration of sympathetic nerve fibers softer different sympathectomy techniques (resection, electrocoagulation and cryodestruction of the part of the sympathetic nerve trunk) following 6 and 12 months. Nine random-bred rabbits were used (Central Research Laboratory, Urals State Medical Academy) to examine the rate of regeneration of the sympathetic nerve trunk contained by the vascular-nervous bundle of rabbit ear. Altogether 18 nerve sympathetic trunks were examined. The rate of regeneration was measured by morphometry. While examining the processes of reparative regeneration of the nerve fibers after resection, electrocoagulation and cryodestruction, account was taken of the changes in their quantity, diameter and degree of myelinization over time. The study of the morphologic picture during comparative analysis of regeneration made on different models of nerve derangement has demonstrated that after resection and electrocoagulation of the sympathetic trunk the process of regeneration proceeds most slowly. On cryogenic injury to the nervous tissue when only nerve fibers are damaged and connective tissue membranes preserve their anatomic integrity, there are created most favourable conditions for regeneration. Toward 12 months after cryodestruction the bundles of nerve fibers acquire an identical diameter, with predominance of the population of thick nerve fibers, and appear mature enough. After electrocoagulation the fibers contained by the bundles are different in diameter, with the predominance of medium-diameter fibers whereas the thick nerve fibers are not many. After resection the thick nerve fibers are lacking whereas the medium-diameter bundles contain myelinated nerve fibers, with the maintenance of the selective decay of the myelin membrane.

Animals↗
PubMed2006

[Sugar beet (Beta vulgaris L.) morphogenesis in vitro: effects of phytohormone type and concentration in the culture medium, type of explants, and plant genotype on shoot regeneration frequency].

In vitro regeneration techniques have been optimized for seven strains and cultivars of sugar beet (Beta vulgaris L.) bred in Russia. The frequency of shoot regeneration from somatic cells and tissues of sugar beet varies from 10 to 97% depending on the explant type, culture-medium composition, and genotype. The in vitro regeneration potential has been estimated in plants with different genotypes. The effect of medium composition (phytohormones and carbohydrates) on the frequency of the formation of a morphogenic callus competent for plant regeneration has been determined. The effect of the types and concentrations of various cytokines (zeatin, kinetin, and 6-benzylaminopurine) on direct shoot regeneration from cotyledon nodes has been estimated. The culture-medium composition has been optimized for direct shoot regeneration from petioles. The effects of different concentrations of abscisic acid on the frequency of shoot regeneration from a morphogenic callus has been studied. Micropropagation has been used to obtain petiole explants and reproduce the shoots obtained by direct regeneration from cotyledon nodes, petioles, and calluses. Improved shoot-regeneration methods can be used for both agrobacterial and bioballistic genetic transformation of the sugar beet genotypes studied.

Beta vulgaris↗
PubMed2006

[Guidance of regenerative axons in optic nerve regeneration in Bcl-2 overexpressing mice].

OBJECTIVE: To determine whether optic nerve can regenerate after optic nerve crush and weather the regenerating axon reaches the target in the brain in Bcl-2 overexpressing mice. METHODS: Left eye optic nerve crush was performed in wild-type (C57Bl/6J) and Bcl-2 transgenic mouse pups at 3 days after birth (P3), followed immediately by the opposite eyeball enucleated. Mouse pups were killed at 4 days post-surgery. Optic nerve regeneration was assessed at the sections of optic nerve and the brain. An anterograde tracer, cholera toxin B subunit conjugated with fluorescein (CTB-F), was applied intraocularly, immediately after optic nerve crush to label retinal ganglion cell axons. Immunofluorescence staining with anti-GAP-43 was carried out to reveal regenerating axons in optic nerve sections. RESULTS: In wild-type mice, severed optic nerves failed to regenerate. In contrast, in all of the Bcl-2 transgenic mice examined, optic nerves regenerated robustly over long distances, but the regenerating fibers were found to deviate from the optic pathway and grew into the forebrain to form aberrant projection. CONCLUSION: When the opposite eyeball is removed in P3 Bcl-2 overexpressing mice with optic nerve was crushed in one side, the guidance from the normal optic pathway is lost, the optic nerve could regenerate and reach the brain, but the regenerating axons could not reach both side of target in the midbrain, they grow into aberrant place of forebrain.

Animals↗
PubMed2006

[Regeneration of optic nerve fibers following graded injuries in rats].

OBJECTIVE: To investigate the changes of retinal ganglion cell (RGC) and their axons, and nerve regeneration ability following graded optic nerve injury (ONI) in rats. METHODS: A pair of cross-action forceps with 148.0 g clipping pressure was used to clip rat optic nerves for 3, 6, 12, 30 and 60 s to induce graded ONI animal model. The RGC was counted at 0.5, 1, 2, 3 and 7 months and the axons were observed 1, 2 and 3 months post-injury. The regeneration process was observed by transmission electron microscopy. The number of optic nerve fibers in transverse sections was calculated in silver-stained longitudinal sections, and a regeneration index (RI) was calculated based on these numbers. The RI, reflecting the regeneration ability of injured optic nerves, was calculated as follows: (number of nerve fibers 0.5 mm behind injury site-number of nerve fibers 2.5 mm behind injury site)/(number of nerve fibers 0.5 mm retrobulbarly-number of nerve fibers 2.5 mm behind injury site). RESULTS: RGC and axons lost continuously after partial ONI. The loss of RGC was fitted with exponential pattern consisted of two phases, acute losing phase within first two weeks post injury and followed by another phase characterized by slowly reducing of RGC. The loss ratio of RGC increased and the survival ratio decreased with the severity of injury intensity. The loss of RGC and axons was aggregated in severe injury and showed a self-limited trend in mild injury. A large amount of clustered, zonal unmyelinated regeneration fibers were present after injury. The RI was 1.409, 1.490, 0.916, 1.119 and 1.224 following 3, 6, 12, 30 and 60 s clipping injury (chi2 = 281.2, P < 0.01), respectively. Different RI was associated to different injury intensity, with a greater regeneration ability in mild injury. CONCLUSIONS: The secondary reaction and regeneration ability vary with graded intensity of optic nerve injury. A self-limited secondary reaction and a more powerful regeneration ability are associated with a mild injury. The repair behavior and the injury may reach a balance and result in a successful regeneration after a certain degree of injury.

Animals↗
PubMed1991

Inflammation near the nerve cell body enhances axonal regeneration.

Although crushed axons in a dorsal spinal root normally regenerate more slowly than peripheral axons, their regeneration can be accelerated by a conditioning lesion to the corresponding peripheral nerve. These and other observations indicate that injury to peripheral sensory axons triggers changes in their nerve cell bodies that contribute to axonal regeneration. To investigate mechanisms of activating nerve cell bodies, an inflammatory reaction was provoked in rat dorsal root ganglia (DRG) through injection of Corynebacterium parvum. This inflammation enhanced regeneration in the associated dorsal root, increasing 4-fold the number of regenerating fibers 17 d after crushing; peripheral nerve regeneration was not accelerated. A milder stimulation of dorsal root regeneration was detected after direct injection of isogenous macrophages into the ganglion. It is concluded that changes favorable to axonal regeneration can be induced by products of inflammatory cells acting in the vicinity of the nerve cell body. Satellite glial cells and other unidentified cells in lumbar DRG were shown by thymidine radioautography to proliferate after sciatic nerve transection or injection of C. parvum into the ganglia. Intrathecal infusion of mitomycin C suppressed axotomy-induced mitosis of satellite glial cells but did not impede axonal regeneration in the dorsal root or the peripheral nerve. Nevertheless, the similarity in reactions of satellite glial cells during 2 processes that activate neurons adds indirect support to the idea that non-neuronal cells in the DRG might influence regenerative responses of primary sensory neurons.

Animals↗
PubMed1990

Ultrastructural comparison between regenerating and developing hindlimbs of Xenopus laevis tadpoles.

Hindlimbs of Xenopus laevis tadpoles at stages 50 to 55 of embryonic development were amputated in order to study the fine structure associated with ontogenetic decline in regenerative ability of this anuran. Regenerating hindlimbs were compared with their contralateral developing limb so as to determine the similarities and differences in ectodermal-mesenchymal ultrastructural relationships in these systems. Prior to stage 53, mesenchymal cells in regenerating limbs, as well as mesenchyme cells in developing limbs appear undifferentiated; the cells are not visibly different. However, at stage 54 muscle and cartilage differentiation in the developing foot is distinct. Furthermore, in all larvae with regenerates at stages 50 to 52 the basal lamina is not evident subjacent to the apical tip of the epithelium which covers the amputation surface. A distinct basal lamina is present, however, beneath the epidermis in all embryonic limbs examined, including the apical tip, as well as in regenerates of stages 53 to 55 tadpoles. Also, a greater apical accumulation of extracellular matrix (ECM) and organized collagen is observed among the mesenchymal cells in regenerates of stages 53 to 55 tadpoles, and among the mesenchyme cells in developing limbs of stages 50 to 55, compared with pre-stage 53 regenerates. In cases in which an embryonic limb bud is composed of relatively undifferentiated cells (stages 50 to 52), events following amputation result in the complete regeneration of a limb. However, when amputated embryonic limbs contain differentiating tissues, (e.g., muscle and cartilage) only selective tissues undergo regeneration resulting in malformed (heteromorphic) regenerates.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Regenerated retinal ganglion cell axons can form well-differentiated synapses in the superior colliculus of adult hamsters.

To investigate in adult animals the distribution and differentiation of the synapses made by axotomized CNS neurons whose regenerating axons are guided back to their natural targets in the brain, we attached an autologous peripheral nerve (PN) graft 2-3 cm in length to the ocular stump of a transected optic nerve (ON) in adult hamsters, inserted the distal end of the graft into the superior colliculus (SC), and, 6-8 weeks later, labeled the retinal ganglion cell (RGC) axons that entered the SC with HRP orthogradely transported from the eye. By light microscopy, regenerated RGC axons extended from the graft into the retinorecipient layers of the SC for up to 500 microns, distances that approximate the lengths of normal RGC arbors. We compared 698 control and 758 regenerated HRP-labeled axon terminals from 4 intact and 4 experimental animals by electron microscopy. The structure of the regenerated RGC terminals, the type of synaptic contacts formed, the ratios of contacts to terminal perimeter, and the domains of the postsynaptic neurons contacted were similar to those of controls. These results indicate that regenerated RGC axons can form well-differentiated synapses in the SC. Morphological differences between the regenerated and control synapses were the larger size of some regenerated terminals, the greater mean length of the regenerated synapses, and the higher proportion of contacts with dendrites that contained vesicles. The synaptic differentiation attained by these reformed retinocollicular projections suggests that regenerating CNS axons and their target neurons in the adult mammalian brain may retain or reexpress certain molecular determinants of normal connectivity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Axonal transport and localization of B-50/GAP-43-like immunoreactivity in regenerating sciatic and facial nerves of the rat.

Neurons that can regenerate their axons following axotomy increase their synthesis and axonal transport of a growth-associated protein, called GAP-43, which has been shown to be identical to the synaptic phosphoprotein B-50. The function of B-50/GAP-43 to the process of regeneration is unknown. We used a polyclonal, affinity-purified antibody against B-50 to study the axonal transport and localization of B-50/GAP-43-like immunoreactivity (B50LI) in the regenerating sciatic and facial nerves of adult rats. Quantitative data were obtained by densitometry of the B-50 band in immunoblots of nerve segments, which had been run on SDS-polyacrylamide gels. In the regenerating sciatic nerve, anterograde accumulation at a collection ligature was 3.0 times higher than retrograde accumulation. The mobile fraction of B50LI was only 0.28 of total B50LI and traveled with a mean anterograde velocity of 5.3 mm/hr. B50LI distribution in the newly regenerated portion of the nerve revealed maximal B50LI levels midway between the position of the crush and the fastest-growing axons. Immunocytochemistry of this portion of the nerve demonstrated B50LI to be associated with regenerating axons but also to a large extent with extra-axonal structures outlining the Schwann cell bands of Büngner. This zone of B50LI-positive Schwann cell bands was found to extend more distally in nerves in which regeneration had processed longer, e.g., up to 5 mm distal to the crush after 3 d and 8 mm after 4 d. Further distal to this zone, many fine regenerating axonal profiles could be detected with B-50 antibody, but were neurofilament negative. These findings raise the possibility of an extra-axonal function of B-50/GAP-43, as this protein might be secreted from regenerating axons and might play a role in axon-Schwann cell interactions during axonal maturation.

Animals↗
PubMed1989

Peripheral nerve regeneration through blind-ended semipermeable guidance channels: effect of the molecular weight cutoff.

Synthetic nerve guidance channels are used to better understand the cellular and molecular events controlling peripheral nerve regeneration. In the present study, the contribution of wound-healing molecules to peripheral nerve regeneration was assessed by varying the molecular weight cutoff of the tubular membrane. Nerve regeneration through polysulfone tubular membranes with molecular weight (Mw) cutoffs of 10(5) and 10(6) Da was analyzed in a transected hamster sciatic nerve model. Cohorts of 6 animals received tubes of either type for 4 or 8 weeks with the distal end of the polymer tube capped. Other cohorts of 6 animals received tubes of either type for 4 weeks with the distal nerve stump secured within the guidance channel so as to create a 4 or 8 mm gap between both nerve stumps. Both types of channels contained regenerated tissue cables extending to the distal end of the guidance channel at both 4 and 8 weeks in the absence of a distal nerve stump. The cables regenerated in the 10(5) Da channels were composed of nerve fascicles surrounded by a loose epineurial sheath, whereas those regenerated in the 10(6) Da channels were composed mainly of granulation tissue. The numbers of myelinated and unmyelinated axons were significantly greater in the 10(5) Da than in the 10(6) Da channels at both 4 and 8 weeks. Both types of channel contained regenerated tissue cables with numerous nerve fascicles when the distal nerve stump was present with either gap length. However, when the gap distance was 8 mm, the 10(6) Da channels contained significantly fewer myelinated axons than the 10(5) Da channels. The present study reveals that the Mw cutoff of a semipermeable guidance channel strongly influences the outcome of peripheral nerve regeneration, possibly by controlling the exchange of molecules between the channel's lumen and the external wound-healing environment. These results suggest that the wound-healing environment secretes humoral factors that can either promote or inhibit the nerve-regeneration process.

Animals↗
PubMed1985

The effects of the fibre environment on the paths taken by regenerating optic nerve fibres in Xenopus.

The paths taken by fibres regenerating to the tectum from various parts of the Xenopus retina were investigated in whole-mount preparations, after localized retinal labelling with HRP. The effects of different environments on the fibres were studied by comparing contralateral with ipsilateral regeneration, in the presence of the other eye or after it had been removed in embryonic life. Under all conditions fibres from the various parts of the retina regenerated to the corresponding appropriate parts of the tectum, but they took a variety of pathways, some grossly abnormal, to get there. Contralaterally regenerating fibres tended to behave less abnormally than ipsilateral fibres; and regeneration in the absence of the other eye tended to be more abnormal than in its presence. In any one category of regeneration the most nearly normal pathways were those of fibres from temporal retina, followed by ventral, nasal and dorsal fibres. Fibres regenerating from all parts of the retina, in the presence of the other eye, tended to become gathered into the medial brachium as they approached the tectum. All regenerating fibres approached their tectal terminations by one or more of three main pathways: round one or both brachia, thus encircling the tectum to get to their terminal zone; directly across the tectum; or by passing on to the tectum before changing course. The changes of direction required to enable fibres wrongly positioned in the tract to reach their correct terminal zones were frequently sudden and considerable, and took place on the tectum or at the tectodiencephalic junction. The results are discussed in relation to the differing substrates over which the fibres regenerate.

Animals↗
PubMed1979

The formation of muscles in regenerating limbs of the newt after denervation of the blastema.

The purpose of this experiment was to examine the relationship, if any, between nerve fibers and the formation of muscle pattern in the regenerating amphibian limb. During embryogenesis, nerve fibers grow into the limb bud at the time when the common muscle blastemas subdivide into individual muscle primordia, whereas in regeneration nerve fibers are always present. In order to learn whether or not the muscle pattern could be laid down in the absence of nerves we amputated 58 limbs of newts (Notophthalmus viridescens) at the mid humeral level and allowed them to regenerate to the medium-bud or late-bud stage. The limbs were then denervated. The majority of limbs denervated at the medium-bud stage either regressed or failed to regenerate further. Regeneration after denervation failed in 9 of 25 limbs denervated at the late-bud stages. In those limbs that continued to regenerate after denervation, the formation of individual muscle primordia did occur, following the same sequence with respect to the gross stage of regeneration as innervated regenerates. In comparing these results with our previous results on the development of muscular pattern in aneurogenic limbs of the axolotl, we conclude that in neither the embryonic nor the regenerating amphibian limb are nerve fibers directly involved in the subdivision of common muscle blastemas into the primordia of individual muscles.

Animals↗