The respective roles of mitotic activity and of cell differentiation in Planarian regeneration

Development ◽  
1968 ◽  
Vol 20 (2) ◽  
pp. 175-188
Author(s):  
Rosine Chandebois

Blastema formation, which is the first step of regeneration in adult Metazoa, is generally considered to be merely an accumulation of undifferentiated cells provided by mitotic activity, which occurs near the wound following amputation. Afterwards, these undifferentiated cells are thought to differentiate rapidly, thus affecting the organization of the regenerate. Some authors have postulated that a close similarity exists between these undifferentiated cells and the blastomeres of a young embryo, and that the influence exercised by the stump tissues in blastema differentiation is a typical inductive process. This concept would imply two successive steps in regeneration: (1) blastema formation, exclusively dependent upon mitotic activity, even after the previous accumulation of undifferentiated cells resulting from migration (Dubois, 1949), (2) the transformation of the blastema into a regenerate as differentiation occurs. Up to now, the first step seemed to be confirmed by experiment, since some authors have observed that some experimental factors which prevent regeneration (X-rays and mitoclastic poisons) inhibit mitoses.

Development ◽  
1984 ◽  
Vol 83 (1) ◽  
pp. 63-80
Author(s):  
Emili Saló ◽  
Jaume Baguñà

Mitotic activity during regeneration in the planarian Dugesia (G) tigrina shows a biphasic pattern, with a first maximum at 4–12 h, a second and higher maximum at 2–4 days, and a relative minimum in between. The first peak is mainly due to pre-existing G2 cells entering mitosis shortly after cutting, whereas the second maximum is due to cells that divide after going through the S period from the onset of regeneration. From a spatial point of view, the highest mitotic values are found in stump (postblastema) regions near the wound (0–300 µm), though regions far from it also show increased mitotic values but always lower overall values. As regeneration continues the postblastema maximum shifts slightly to more proximal regions. In contrast, no mitosis has been found within the blastema, even though the number of blastema cells increases steadily during regeneration. These results suggest that blastema in planarians forms through an early accumulation of undifferentiated cells at the wound boundary, and grows by the continuous local migration of new undifferentiated cells from the stump to the base of blastema. The results obtained demonstrate that blastema formation in planarians occurs through mechanisms somewhat different to those shown to occur in the classical epimorphic models of regeneration (Annelida, Insecta, Amphibia), and suggest that planarian regeneration could represent an intermediate stage between morphallactic and epimorphic modalities of regeneration.


Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 29-36
Author(s):  
V. Gremigni ◽  
M. Nigro ◽  
I. Puccinelli

The source and fate of blastema cells are important and still unresolved problems in planarian regeneration. In the present investigation we have attempted to obtain new evidence of cell dedifferentiation-redifferentiation by using a polyploid biotype of Dugesia lugubris s.1. This biotype is provided with a natural karyological marker which allows the discrimination of triploid embryonic and somatic cells from diploid male germ cells and from hexaploid female germ cells. Thanks to this cell mosaic we previously demonstrated that male germ cells take part in blastema formation and are then capable of redifferentiating into somatic cells. In the present investigation sexually mature specimens were transected behind the ovaries and the posterior stumps containing testes were allowed to regenerate the anterior portion of the body. Along with the usual hexaploid oocytes, a small percentage (3.2%) of tetraploid oocytes were produced from regenerated specimens provided with new ovaries. By contrast only hexaploid oocytes were produced from control untransected specimens. The tetraploid oocytes are interpreted as original diploid male germ cells which following the transection take part in blastema formation and then during regeneration redifferentiate into female germ cells thus doubling their chromosome number as usual for undifferentiated cells entering the female gonad in this biotype.


Development ◽  
1985 ◽  
Vol 89 (1) ◽  
pp. 57-70
Author(s):  
Emili Saló ◽  
Jaume Baguñà

One of the tenets of Wolff and Dubois' ‘neoblast theory’ of planarian regeneration (Wolff & Dubois, 1948) is that blastema is mainly formed by the accumulation of undifferentiated parenchymal cells (neoblasts) that can migrate, if needed, over long distances to the wound. That neoblasts migrate was claimed by these authors after partial X-irradiation, and total Xirradiation and grafting using planarian strains of different pigmentation. From this they suggested that migration of neoblasts is stimulated by the wound and directed towards it. To study the nature and extent of such ‘migration’ in intact and regenerating organisms, and in order to avoid the flaws of using pigmentation as a marker, we made grafts between sexual and asexual races of Dugesia(S)mediterranea that differ in a chromosomal marker, and between diploid and tetraploid biotypes of Dugesia(S)polychroa that differ in nuclear size. Also, fluorescent latex beads were used as cytoplasmic markers to follow ‘migration’ of differentiated cells. The hosts were irradiated or non-irradiated intact and regenerating organisms. The results show that: 1) movement of graft cells into host tissues occurs in intact organisms at a rate of ≃40µm/day, and that this increases up to ≃75µm/day in irradiated hosts; 2) movement of cells occurs evenly in all directions; 3) regeneration does not speed up rate of movement nor drives cells preferentially to the wound; 4) spreading of cells is mainly due to the movement of undifferentiated cells (neoblasts); and 5) higher rates of movement are correlated with higher mitotic indexes. From this, it is concluded that the so-called ‘migration’ of neoblasts is not a true cell migration but the result of the slow, even and progressive spreading of these cells mainly caused by random movements linked to cell proliferation. The implications of these results for blastema formation and the origin of blastema cells are discussed.


Development ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 53-63
Author(s):  
V. Gremigni ◽  
C. Miceli ◽  
I. Puccinelli

Specimens from a polyploid biotype of Dugesia lugubris s.l. were used to clarify the role and fate of germ cells during planarian regeneration. These specimens provide a useful karyological marker because embryonic and somatic cells (3n = 12) can be easily distinguished from male (2n = 8) and female (6n = 24) germ cells by their chromosome number. We succeed in demonstrating how primordial germ cells participate in blastema formation and take part in rebuilding somatic tissues. This evidence was obtained by cutting each planarian specimen twice at appropriate levels. The first aimed to induce primordial germ cells to migrate to the wound. The second cut was performed after complete regeneration and aimed to obtain a blastema from a cephalic or caudal area devoid of gonads. A karyological analysis of mitotic cells present in each blastema obtained after the second cut provided evidence that cells, originally belonging to the germ lines, are still present in somatic tissues even months after complete regeneration. The role of primordial germ cells in planarian regeneration was finally discussed in relation to the phenomenon of metaplasia or transdifferentiation.


Development ◽  
1989 ◽  
Vol 107 (1) ◽  
pp. 69-76 ◽  
Author(s):  
E. Salo ◽  
J. Baguna

In planarians, blastema cells do not divide, and growth blastema is thought to result from the steady wound epithelium, of undifferentiated cells produced in the stump. However, whether these cells come only sources or whether cells placed far from the wound can participate, after long-range migrations, in the still uncertain. To study this problem, we have parameters of the process of regeneration: cell growth; number of cells produced by mitosis in the wound (postblastema); and rates of movement undifferentiated cells using grafting procedures with chromosomal markers. The results show that: (1) cells area spread (move) at higher rates than cells placed (90–140_mday-1 versus 40–50_mday-1); (2) cells than 500_m from the wound boundary are hardly 5-day-old blastemata; and (3) the number of cells within a 200–300_m postblastema area around the wound explain, provided their rates of movement are taken increasing number of blastema cells. From this, it is blastema cells in planarians originate from local mitotic activity jointly with local cell movement postblastema area around the wound match the blastema cells during regeneration. The implications for blastema growth and pattern formation mechanisms


2001 ◽  
Vol 79 (7) ◽  
pp. 1171-1208 ◽  
Author(s):  
M C Thorndyke ◽  
MD Candia Carnevali

There has been much recent interest in the presence and biological functions of growth regulators in invertebrates. In spite of the different distribution patterns of these molecules in different phyla (from molluscs, insects, and annelids to echinoderms and tunicates), they seem always to be extensively involved in developmental processes, both embryonic and regenerative. Echinoderms are well known for their striking regenerative potential and many can completely regenerate arms that, for example, are lost following self-induced or traumatic amputation. Thus, they provide a valuable experimental model for the study of regenerative processes from the macroscopic to the molecular level. In crinoids as well as probably all ophiuroids, regeneration is rapid and occurs by means of a mechanism that involves blastema formation, known as epimorphosis, where the new tissues arise from undifferentiated cells. In asteroids, morphallaxis is the mechanism employed, replacement cells being derived from existing tissues following differentiation and (or) transdifferentiation. This paper focuses on the possible contribution of neurohormones and growth factors during both repair and regenerative processes. Three different classes of regulatory molecules are proposed as plausible candidates for growth-promoting factors in regeneration: neurotransmitters (monoamines), neuropeptides (substance P, SALMFamides 1 and 2), and growth-factor-like molecules (TGF-β (transforming growth factor β), NGF (nerve growth factor), RGF-2 (basic fibroblast growth factor)).


1941 ◽  
Vol 25 (2) ◽  
pp. 275-291 ◽  
Author(s):  
A. Marshak

1. A method for isolating nuclei in quantity from mammalian tissues is described. 2. The rate of uptake of radioactive phosphorus by nuclei is found to be quite rapid. The phosphorus was shown not to be taken up by exchange. 3. Nuclei of tumors accumulate more radioactive phosphorus than normal liver nuclei. This was shown to be due to mitotic activity and not a form of metabolism peculiar to tumor cells. 4. The specific activities of nuclei and cytoplasm are compared. 5. 60 to 70 per cent of the nuclear radioactive phosphorus is present as nucleoprotein from 1 hour to 5 days after it is administered. In the lymphoma nuclei 90–95 per cent of the phosphorus is in the nucleoprotein fraction from 1–5 days after it is administered. 6. The specific activities of the nucleoprotein, lipid, and acid-soluble fractions of liver and tumor nuclei are compared. 7. From the rate of P32 uptake by nuclei it is calculated that a new lymphoma nucleus is synthesized on the average once every 27 hours. This is in agreement with the observed rate of growth of the tumor. 8. In the lymphoma nucleus it is calculated that 7 x 104 molecules of tetranucleotide are synthesized per second. 9. Irradiation with 200 r. x-rays alters the distribution of P32 in the lymphoma cell, markedly increasing the concentration in the nucleus shortly after irradiation. The P32 concentration in the cytoplasm decreases with time after irradiation. It is suggested that the altered distribution is correlated with the inhibition of mitosis produced by the x-rays. 10. Continual synthesis of nucleoprotein takes place even in nuclei of cells which do not undergo mitosis.


Development ◽  
1996 ◽  
Vol 122 (5) ◽  
pp. 1567-1575 ◽  
Author(s):  
S.E. Clark ◽  
S.E. Jacobsen ◽  
J.Z. Levin ◽  
E.M. Meyerowitz

The CLAVATA (CLV1 and CLV3) and SHOOT MERISTEMLESS (STM) genes specifically regulate shoot meristem development in Arabidopsis. CLV and STH appear to have opposite functions: c1v1 and Clv3 mutants accumulate excess undifferentiated cells in the shoot and floral meristem, while stm mutants fail to form the undifferentiated cells of the shoot meristem during embryonic development. We have identified a weak allele of stm (stm-2) that reveals STM is not only required for the establish- ment of the shoot meristem, but is also required for the continued maintenance of undifferentiated cells in the shoot meristem and for proper proliferation of cells in the floral meristem. We have found evidence of genetic interactions between the CLV and STM loci. clv1 and c1v3 mutations partially suppressed the stm-1 and stm-2 phenotypes, and were capable of suppression in a dominant fashion. clv stm double mutants and plants homozygous for stm but heterozygous for clv, while still lacking an embryonic shoot meristem, exhibited greatly enhanced postembryonic shoot and floral meristem development. Although stm phenotypes are recessive, stm mutations dominantly suppressed clv homozygous and heterozygous phenotypes. These results indicate that the stm phenotype is sensitive to the levels of CLV activity, while the clv phenotype is sensitive to the level of STM activity. We propose that these genes play related but opposing roles in the regulation of cell division and/or cell differentiation in shoot and floral meristems.


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