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[Cytochemical analysis of muscle cell histones of Triturus cristatus limbs a) in normal regeneration, b) in regeneration arrested by r-irradiation, and c) in regeneration restored by implants].

The muscular cells of regenerating limbs and of limbs in which regenerative power is restored, show an important decrease in the amount of cytophotometrically detected histones. This decrease is owing to the arginine rich fraction and to the lysine rich fraction. The muscular cells of irradiated limbs show a decrease in the amount of histones. This decrease is owing only to the arginine rich fraction and continues after the thirieth day of irradiation and amputation.

Animals

Leg regeneration in the cockroach, Blatella germanica. II. Regeneration from a non-congruent tibial graft/host junction.

The interactions occurring between host and graft leg epidermis at a non-congruent junction were studied in the cockroach, Blatella germanica. Graft and host tibia were cut perpendicular to the proximal-distal axis and two heteropleural combinations were used to reverse separately the two transverse axes of the graft relative to the host. Use of dark and light cuticle colour mutants gave a good indication of the graft or host origin of regenerated structures. Graft/host junctions regenerated segmented structures in various spatial arrangements, always comprising two copies of all structures distal to the level of the junction. It is concluded that the categories--two separate laterals, double lateral, completely and partially autonomous regeneration--reflect two processes. (i) If the graft tarsus is removed, graft and host may not heal together and interact, but form autonomous regenerates lying in mirror-image symmetry separating original graft and host levels. (ii) If interaction occurs between graft and host (or their developing autonomous regenerates) two laterals of dual origin are produced, one from each point of transverse axis incongruity. These laterals may secondarily fuse together to form a double structure originating from a point of congruity. The orientation and composition of the component tarsi of the double structure depend on the site of origin and the extent to which the two laterals fuse. It is argued that the four 'faces' and two 'transverse axes' of the leg are merely descriptive terms. A new model is developed whereby lateral regeneration arises directly from the circumferential organisation of the leg epidermis. Previous work has shown that position is specified continuously around the circumference, and that intercalary regeneration occurs by the shortest route between confronted positions. After reversal of one 'transverse axis' the shortest route between confronted graft and host positions is different on the two sides of each of the two points of 'axis' incongruity, and at these points the two halves of a complete circumference are formed. These lateral circumferences, like the terminal circumference exposed by amputation, cannot heal over by intercalary regeneration, and this leads to regeneration of distal structures. The model accounts for lateral regeneration after reversal of both 'transverse axes' by 180 degrees rotation of a homopleural graft. The possibility is discussed that there may be clonal restrictions on the circumferential positions which the progeny of a cell may occupy.

Animals

Kinetics of the regeneration of sea-urchin cilia. II. Regeneration of animalized cilia.

The kinetics of the regeneration of cilia of Arbacia punctulata animalized by treatment with trypsin have been examined. The cilia regenerate with biphasic kinetics: an initial linear phase which changes abruptly to a second, slower linear rate. Inhibition of protein synthesis with 10 microM emetine has little effect on the first phase but totally abolishes the second, suggesting that the first phase represents the utilization of a pre-existing precursor pool while the second reflects de novo protein synthesis. However, cilia will regenerate again following a second deciliation in the presence of emetine, to half the plateau value of the first regeneration, indicating that only a fraction of the pool of the limiting precursor can be assembled into the regenerating cilium. It is proposed that a residual pool of this precursor is required to maintain the assembly-disassembly equilibrium in favour of the assembled organelle.

Animals

Quantitative studies of the regeneration of rat myelinated nerve fibres: variations in the number and size of regenerating fibres after repeated localized freezings.

The number and size of myelinated nerve fibres were determined in the nerve to the medial head of the gastrocnemius muscles of rats whose left sciatic nerve was repeatedly frozen (one to five times at three weekly intervals). The contralateral nerve was used as a control. Results varied according to the number of freezings performed and, for a given number of freezings, according to the period of regeneration. When measurements were completed 1 month after the last of several localized freezings, the number of regenerating myelinated nerve fibres increased regularly up to the third freezing, reaching to about 220% of the control value, but no higher values were recorded after four or five freezings. The nerve fibre distribution was unimodal in all the nerves studied. The mean diameter of all myelinated fibres decreased with the number of freezings from 50% of the control value after the first to 36% after the fifth. When measurements were made 1, 3, 6, 12 and 18 months after the third and final freezing, the number of regenerating myelinated nerve fibres decreased by about 30% between the first and third month and then stabilized at 190% of the control value. Nerve fibre distribution became bimodal from the third month onwards, and the mean diameter of all myelinated fibres increased regularly. However, by the eighteenth month, the size of regenerated myelinated nerve fibres had only reached 70% of the normal contralateral value.

Animals

[Regeneration of a whole worm from a small fragment of the body of Dugesia tigrina planaria following repeated removal of regenerates].

The ability of restoration of the whole worm from a small body fragment was not reduced after 8 removals of the anterior and posterior regenerates. The polarity of restoring fragments in the most cases corresponded to the initial one. Upon the colchicine effect on the restoring fragments which accompanied the repeated removal of the regenerates, a high percentage of cases of incomplete somatic embryogenesis was observed: the formation of bi- and multipolar forms. The formation of regenerates is realized by neoblasts. Upon the repeated removal of regenerates, the dedifferentiated intestine cells appear to take part in the restorative processes as well.

Animals

Hormone control in regeneration: effects of somatostatin on appendage regeneration, blood glucose and liver glycogen in Diemictylus viridescens.

In the present communication, synthetic somatostatin, a hypothalamic factor which has a known inhibitory effect on the release of growth hormone, thyroid-stimulating hormone, prolactin, insulin and glucagon in man and other mammals, was found to have an inhibitory effect on limb and tail regeneration in adult Diemictylus viridescens, when the newts were treated with a daily dose of 3-5 or 15 microgram/animal for a period of 34 days post-amputation. At the higher dose, the animals exhibited total inhibition of appendage regeneration in a few cases and the remainder showed a considerable delay compared to the controls; none of the experimental animals reached the advanced four-digit stage achieved by the controls. Furthermore, the blood glucose and liver glycogen values in the somatostatin-treated animals were significantly lower than the control values. Mechanisms in the storage, mobilization and utilization of glucose (involving hormones) are discussed in relation to appendage regeneration in the newt and possible controls of regeneration at the level of the hypothalamus are suggested.

Animals

Bioelectricity and regeneration: large currents leave the stumps of regenerating newt limbs.

Electrical currents near regenerating newt limbs were measured with a recently developed vibrating probe. Steady currents with local surface densities of 10 to 100 muA/cm2 or more leave the end of the stump during the first 5-10 days after amputation and are balanced by currents with densities of only 1-3 muA/cm2 that enter the intact skin around the stump. They are immediately dependent upon the entry of sodium ions into this skin and are therefore inferred to be skin-driven. The outward currents are comparable in direction, density, duration, and position to artificially imposed currents previously found sufficient to induce significant regeneration of amputated adult frog limbs. This comparison suggests that the endogenous stump currents play some causal role in initiating regeneration.

Amiloride

[Ultrastructure of the cells and DNA synthesis in skeletal muscle regeneration. A study of the regeneration of the frog sartorius muscle by an electron microscopic autoradiographic method].

The ultrastructure of cells of the regenerating frog's sartorius muscle and their capacity to synthesize DNA was studied by means of 3H-thymidine (3HT) electron microscope autoradiography. On the 8-17th post injury (p.i.) days, 2 hours following 3HT administration, only mononuclear cells were seen labeled, the myotube nuclei incorporating no 3HT. Along with the endothelial cells, fibroblasts, phagocytes and cells identified conventionally as myoblasts, satellite cells examined from both necrotic and viable parts of injured myofibers were labeled. No myoblast sequestration from the injured myofibers occurred. By the 13-15th p.i. days, numerous myoblast-like cells are accumulated beneath the glycocalix layer covering the free ends of myotubes which are rich in ribosomes and display an active sarcomerogenesis. Some of these myoblast-like cells become labeled after 3HT pulse. The 13 day p.i. regenerates examined 72 hours following 3HT injection display labeling in numerous myotube nuclei. This is indicative of the myoblast fusion, which is believed to play a principal role in the regenerative somatic myogenesis. Within the myonuclei adjacent to the areas of the regeneration, membranous and/or fibrillar structures of an unknown origin were frequently observed.

Animals

[Variations in the completeness of skin regeneration during post-traumatic regeneration].

The healing of the full-thickness skin wounds on the abdomen and the back of hedgehogs was investigated. The wound healing on the abdomen proved to result mainly from the wound contraction. In most cases a small regenerated area containing new hair and sebaceous glands was formed in the centre of the wound. Contraction of the wound was practically absent on the back and its healing occurred chiefly by new tissue formation; due to this vast regenerated areas were formed in the wound. Connective tissue frame-work of the regenerated area on the abdomen and the back resembled intact derma by fiber distribution.

Animals

[Quantitative studies on the regeneration of myelinated nerve fibers. II. Variations of the number and size of regenerating nerve fibers after localized crushing or total section].

1. The number and size of myelinated nerve fibers have been determined at standard levels, in the nerve to medial head of right and left gastrocnemius muscles of 112 rats in which the left sciatic nerve had suffered an experimental lesion according one of the following four modalities: localized crushing, total section followed or not by suture and resection of a nervous segment of about 1 cm. 2. In the nerve to medial head of right gastrocnemius muscle (contralateral nerve used as control), the number of myelinated fibers decreased in average to 10% after crushing, 5% or 4% after section followed or not by suture. However, an increase of 6% was observed after resection. The mean values of the mean diameters showed a decrease of 8% after crushing and 5% after section without suture. This value did not seem to be affected by section followed by immediate suture and after resection, it increased of 11%. On the whole, male rats appeared to be more sensitive than female to the effects of the operation. 3. The nerves of 12 rats have been observed from 15 to 334 days after resection of about 1 cm of sciatic nerve. The 20% of the regenerating myelinated nerve fibers which have succeeded to cross over such a distance had a distribution which remained unimodal; the diameter of the large fibres did not exceed 8 micronm. 4. 34 rats have been sacrificed from 15 to 715 days after sciatic nerve section which was not followed by suture. The number of myelinated nerve fibers became normal again during the 4th month and reached afterwards a mean value of 130%, with very marked variations. The nerve fibre distribution was most frequently unimodal, but may came bimodal one year after the operation in certain nerves. Their mean diameter never exceeded 60% of the normal. 5. The nerves of 34 rats have been examined from 15 to 720 days after section and immediate suture. The number of myelinated nerve fibers returned to normal during the second month and increased afterwards to an average of 150% with very important variations. The nerve fiber distribution was generally unimodal, but may become bimodal 7 months after the operation. Their mean diameter reached only 50 to 55% of the normal. 6. 32 rats have been sacrificed from 10 to 720 rats after a localized crushing. The number of myelinated nerve fibers come back to normal during the 4th week and later increased up to a mean of 115%. Their distribution became early bimodal from the 97th day onwards. Although, their mean diameter nerver exceeded 80% of the normal, the histograms of the regenerating nerve and of the control nerve could be almost superposed during the second year.

Animals

[Eyestalk removal and intensive regeneration: joint effects on the molting cycle of Pachygrapsus marmoratus (Crustacea, Decapoda), variability of the critical regeneration stage].

When eyestalk-removal and pereiopod-removal, followed by intensive regeneration, are realized together on the same animal, their accelerative effect on the moulting rate is lower than the effect of only one of these two processes. The critical stage of pereiopod regeneration varies between the moulting stages C4 to D1 according to the physiological state of the Crab.

Animals

[Quantitative studies on the regeneration of myelinated nerve fibers. III. Variations in the number and size of regenerated fibers after localized freezing].

The number and size of myelinated nerve fibers have been determined at standard levels in the nerve to medial head of right and left gastroenemius muscles of 45 rats in which the left sciatic nerve had suffered an localized freezing. The nerves have been observed from 10 to 720 days after the operation. In the contralateral nerve, the number of myelinated fibers decreased in average to 18.8%, the mean diameter to 11,2%. Male rats appeared to be more sensitive than female to the effects of the operation. In the regenerating nerve, the number of myelinated fibers returned to normal during 4th week and later increased up to a mean of 124%. Their distribution became bimodal after the 60th day. Both histograms could be superposed by the 330th day and the mean diameter came back to normal soon afterwards.

Animals