An official website of the United States government
Abstract An animal's ability to regrow lost tissues or structures can vary greatly during its life cycle. The annelidCapitella teletaexhibits posterior, but not anterior, regeneration as juveniles and adults. In contrast, embryos display only limited replacement of specific tissues. To investigate when during development individuals ofC. teletabecome capable of regeneration, we assessed the extent to which larvae can regenerate. We hypothesized that larvae exhibit intermediate regeneration potential and demonstrate some features of juvenile regeneration, but do not successfully replace all lost structures. Both anterior and posterior regeneration potential of larvae were evaluated following amputation. We used several methods to analyze wound sites: EdU incorporation to assess cell proliferation; in situ hybridization to assess stem cell and differentiation marker expression; immunohistochemistry and phalloidin staining to determine presence of neurites and muscle fibers, respectively; and observation to assess re‐epithelialization and determine regrowth of structures. Wound healing occurred within 6 h of amputation for both anterior and posterior amputations. Cell proliferation at both wound sites was observed for up to 7 days following amputation. In addition, the stem cell markervasawas expressed at anterior and posterior wound sites. However, growth of new tissue was observed only in posterior amputations. Neurites from the ventral nerve cord were also observed at posterior wound sites. De novoashexpression in the ectoderm of anterior wound sites indicated neuronal cell specification, although the absence ofelavexpression indicated an inability to progress to neuronal differentiation. In rare instances, cilia and eyes re‐formed. Both amputations induced expanded expression of the myogenesis geneMyoDin preexisting tissues. Our results indicate that amputated larvae complete early, but not late, stages of regeneration, which indicates a gradual acquisition of regenerative ability inC. teleta. Furthermore, amputated larvae can metamorphose into burrowing juveniles, including those missing brain and anterior sensory structures. To our knowledge, this is the first study to assess regenerative potential of annelid larvae.
more »
« less
Tail regeneration in a cephalochordate, the Bahamas lancelet, Asymmetron lucayanum
Abstract Lancelets (Phylum Chordata, subphylum Cephalochordata) readily regenerate a lost tail. Here, we use light microscopy and serial blockface scanning electron microscopy (SBSEM) to describe tail replacement in the Bahamas lancelet,Asymmetron lucayanum. One day after amputation, the monolayered epidermis has migrated over the wound surface. At 4 days, the regenerate is about 3% as long as the tail length removed. The re‐growing nerve cord is a tubular outgrowth of ependymal cells, and the new part of the notochord consists of several degenerating lamellar cells anterior to numerous small vacuolated cells. The cut edges of the mesothelium project into the regenerate as tubular extensions. These tubes anastomose with each other and with midline mesodermal canals beneath the regenerating edges of the dorsal and ventral fins. SBSEM did not reveal a blastema‐like aggregation of undifferentiated cells anywhere in the regenerate. At 6 days, the regenerate (10% of the amputated tail length) includes a notochord in which the small vacuolated cells mentioned above are differentiating into lamellar cells. At 10 days, the regenerate is 22% of the amputated tail length: myocytes have appeared in the walls of the myomeres, and sclerocoels have formed. By 14 days, the regenerate is 35% the length of the amputated tail, and the new tissues resemble smaller versions of those originally lost. The present results forA. lucayanum, a species regenerating quickly and with little inter‐specimen variability, provide the morphological background for future cell‐tracer, molecular genetic, and genomic studies of cephalochordate regeneration.
more »
« less
- Award ID(s):
- 1952567
- PAR ID:
- 10454712
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Journal of Morphology
- Volume:
- 282
- Issue:
- 2
- ISSN:
- 0362-2525
- Format(s):
- Medium: X Size: p. 217-229
- Size(s):
- p. 217-229
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Abstract Background The cellular basis of adult growth in cephalochordates (lancelets or amphioxus) has received little attention. Lancelets and their constituent organs grow slowly but continuously during adult life. Here, we consider whether this slow organ growth involves tissue-specific stem cells. Specifically, we focus on the cell populations in the notochord of an adult lancelet and use serial blockface scanning electron microscopy (SBSEM) to reconstruct the three-dimensional fine structure of all the cells in a tissue volume considerably larger than normally imaged with this technique. Results In the notochordal region studied, we identified 10 cells with stem cell-like morphology at the posterior tip of the organ, 160 progenitor (Müller) cells arranged along its surface, and 385 highly differentiated lamellar cells constituting its core. Each cell type could clearly be distinguished on the basis of cytoplasmic density and overall cell shape. Moreover, because of the large sample size, transitions between cell types were obvious. Conclusions For the notochord of adult lancelets, a reasonable interpretation of our data indicates growth of the organ is based on stem cells that self-renew and also give rise to progenitor cells that, in turn, differentiate into lamellar cells. Our discussion compares the cellular basis of adult notochord growth among chordates in general. In the vertebrates, several studies implied that proliferating cells (chordoblasts) in the cortex of the organ might be stem cells. However, we think it is more likely that such cells actually constitute a progenitor population downstream from and maintained by inconspicuous stem cells. We venture to suggest that careful searches should find stem cells in the adult notochords of many vertebrates, although possibly not in the notochordal vestiges (nucleus pulposus regions) of mammals, where the presence of endogenous proliferating cells remains controversial.more » « less
-
Abstract Many species in the tunicate family Molgulidae have independently lost their swimming larval form and instead develop as tailless, immotile larvae. These larvae do not develop structures that are essential for swimming such as the notochord, otolith, and tail muscles. However, little is known about neural development in these nonswimming larvae. Here, we studied the patterning of the Motor Ganglion (MG) ofMolgula occulta, a nonswimming species. We found that spatial patterns of MG neuron regulators in this species are conserved, compared with species with swimming larvae, suggesting that the gene networks regulating their expression are intact despite the loss of swimming. However, expression of the key motor neuron regulatory geneEbf (Collier/Olf/EBF)was reduced in the developing MG ofM. occultawhen compared with molgulid species with swimming larvae. This was corroborated by measuring allele‐specific expression ofEbfin hybrid embryos from crosses ofM. occultawith the swimming speciesM. oculata. Heterologous reporter construct assays in the model tunicate speciesCiona robustarevealed a specificcis‐regulatory sequence change that reduces expression ofEbfin the MG, but not in other cells. Taken together, these data suggest that MG neurons are still specified inM. occultalarvae, but their differentiation might be impaired due to reduction ofEbfexpression levels.more » « less
-
Abstract Birds are known to act as potential vectors for the exogenous dispersal of bryophyte diaspores. Given the totipotency of vegetative tissue of many bryophytes, birds could also contribute to endozoochorous bryophyte dispersal. Research has shown that fecal samples of the upland goose (Chloephaga picta) and white‐bellied seedsnipe (Attagis malouinus) contain bryophyte fragments. Although few fragments from bird feces have been known to regenerate, the evidence for the viability of diaspores following passage through the bird intestinal tract remains ambiguous. We evaluated the role of endozoochory in these same herbivorous and sympatric bird species in sub‐Antarctic Chile. We hypothesized that fragments of bryophyte gametophytes retrieved from their feces are viable and capable of regenerating new plant tissue. Eleven feces disk samples containing undetermined moss fragments fromC. picta(N = 6) andA. malouinus(N = 5) and six moss fragment samples from wild‐collected mosses (Conostomumtetragonum,Syntrichiarobusta, andPolytrichumstrictum) were grown ex situ in peat soil and in vitro using a agar Gamborg medium. After 91 days, 20% of fragments fromA. malouinusfeces, 50% of fragments fromC. pictafeces, and 67% of propagules from wild mosses produced new growth. The fact that moss diaspores remained viable and can regenerate under experimental conditions following the passage through the intestinal tracts of these robust fliers and altitudinal and latitudinal migrants suggests that sub‐Antarctic birds might play a role in bryophyte dispersal. This relationship may have important implications in the way bryophytes disperse and colonize habitats facing climate change.more » « less
-
null (Ed.)Cells often respond to changes in their environment by producing new molecules and building new cell components, such as proteins, which perform most tasks in the cell, or DNA and RNA, which carry genetic information. Complex tissues – such as limbs, which are made up of muscles, tendons, bones and cartilage – are difficult to see through, so studying when and where cells in these tissues produce different types of molecules is challenging. New approaches combining advanced three-dimensional microscopy and fluorescent labelling of molecules could provide a way to study these processes within whole animal tissues. One application for this is studying how salamanders regrow lost limbs. When salamanders such as axolotls regrow a limb, some cells in the limb stump form a group called the blastema. The blastema contains cells that are specialized to different purposes. Each cell in the blastema produces many new proteins as well as new DNA and RNA molecules. Fluorescently labeling particular molecules and taking images of the regenerating limb at different times can help to reveal how these new molecules control and coordinate limb regrowth. Duerr et al. developed a three-dimensional microscopy technique to study the production of new molecules in regenerating axolotl limbs. The method labeled molecules of different types with fluorescent markers. As a result, new proteins, RNA and DNA glowed under different colored lights. Duerr et al. used their method to show that nerve damage, which hinders limb regrowth in salamanders, reduces DNA production in the blastema. There are many possible applications of this microscopy method. Since the technique allows the spatial arrangement of the cells and molecules studied to be preserved, it makes it possible to investigate which molecules each cell is making and how they interact across a tissue. Not only does the technique have the potential to reveal much more about limb regrowth at all stages, but the fluorescent markers used can also be easily adapted to many other applications.more » « less
