Cell movement in intact and regenerating planarians. Quantitation using chromosomal, nuclear and cytoplasmic markers

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 ◽  
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 ◽  
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


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 ◽  
1989 ◽  
Vol 107 (1) ◽  
pp. 77-86 ◽  
Author(s):  
J. Baguna ◽  
E. Salo ◽  
C. Auladell

In most regenerating systems, blastema cells arise by dedifferentiation of functional tissue cells. In is still debatable whether dedifferentiated cells or a undifferentiated cells, the neoblasts, are the main cells. Moreover, it is unclear whether in the intact neoblasts are quiescent cells ‘reserved’ for serve as functional stem cells of all differentiated uncertainties partly stem from the failure to conventional labelling methods neoblasts from Here we describe a new approach to these problems regenerative and stem cell capabilities of purified differentiated cells when introduced, separately, into hosts. Introduction of neoblasts led to resumed blastema formation, and extended or complete survival differentiated cells, in contrast, never did so. neoblasts can be qualified as totipotent stem cells of blastema cells, while dedifferentiation does not either in intact or regenerating organisms. In strengthen the idea that different types of formation, linked to the tissular complexity of the present in the animal kingdom.


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.


2021 ◽  
Vol 11 (9) ◽  
pp. 3729
Author(s):  
Katarzyna Balon ◽  
Benita Wiatrak

Models based on cell cultures have become a useful tool in modern scientific research. Since primary cell lines are difficult to obtain and handle, neoplasm-derived lines like PC12 and THP-1 offer a cheap and flexible solution for neurobiological studies but require prior differentiation to serve as a neuronal or microglia model. PC12 cells constitute a suitable research model only after differentiation by incubation with nerve growth factor (NGF) and THP-1 cells after administering a differentiation factor such as phorbol 12-myristate-13-acetate (PMA). Still, quite often, studies are performed on these cancer cells without differentiation. The study aimed to assess the impact of PC12 or THP-1 cell differentiation on sensitivity to harmful factors such as Aβ25-35 (0.001–5 µM) (considered as one of the major detrimental factors in the pathophysiology of Alzheimer’s disease) or lipopolysaccharide (1–100 µM) (LPS; a pro-inflammatory factor of bacterial origin). Results showed that in most of the tests performed, the response of PC12 and THP-1 cells induced to differentiation varied significantly from the effect in undifferentiated cells. In general, differentiated cells showed greater sensitivity to harmful factors in terms of metabolic activity and DNA damage, while in the case of the free radicals, the results were heterogeneous. Obtained data emphasize the importance of proper differentiation of cell lines of neoplastic origin in neurobiological research and standardization of cell culture handling protocols to ensure reliable results.


1986 ◽  
Vol 6 (11) ◽  
pp. 3920-3927
Author(s):  
K Ariizumi ◽  
H Ariga

A small circular DNA was found extrachromosomally in a clone of F9 embryonal carcinoma (EC) cells at high copy numbers per cell. The DNA was cloned in plasmid pUC19. Restriction endonuclease analyses of the DNA indicated that the DNA (fPyF9) was a mutant of polyomavirus (Py) DNA and had a mutation in a noncoding regulatory region. There have been many reports on the isolation of Py mutants capable of replication in undifferentiated cells. However, fPyF9 was different from other Py mutants in the following aspects: it was harbored stably as a free copy at 1 X 10(4) to 5 X 10(4) copies per cell in EC cells; it replicated in undifferentiated cells better than in differentiated cells; it was extremely rearranged in the sequences of the enhancer B domain; and it carried in the enhancer B domain three copies of an exogenous sequence which does not exist in Py strain A2. From these observations, we propose a new class of Py EC mutant which has an autonomous state similar to that of plasmid and small circular DNA in host cells.


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.


2018 ◽  
Author(s):  
Víctor Alejandro Zapata Trejo

The epigenome regulates the gene expression of all differentiated cells and indicates which specific genes must be transcribed. It is argued that the expression factors that act on specific genes of the somatic cell involved in a behavior also act on the transcription of the same genes in the most undifferentiated cells of the germ line. It is proposed how a probabilistic view of the random mutation can explain the evolution of the phenotypes and integrate all the evidence pointing to a joint evolution with the environment.


2018 ◽  
Author(s):  
Víctor Alejandro Zapata Trejo

The epigenome regulates the gene expression of all differentiated cells and indicates which specific genes must be transcribed. It is argued that the expression factors that act on specific genes of the somatic cell involved in a behavior also act on the transcription of the same genes in the most undifferentiated cells of the germ line. It is proposed how a probabilistic view of the random mutation can explain the evolution of the phenotypes and integrate all the evidence pointing to a conjunct evolution with the environment.


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