Biology subjects

Bolanos, D. M.

Publications and source records attributed to Bolanos, D. M..

3 recordsLinked to original sources

bioRxiv2024

Reproductive life history of an acoel worm

Acoel worms belong to an enigmatic and understudied animal lineage in the phylum Xenacoelomorpha. Sparse taxonomic and histological work suggests that these worms exhibit a diversity of reproductive anatomies and likely a corresponding diversity in reproductive behavior. Here, we study the reproductive life history of the three-banded panther worm Hofstenia miamia, an acoel that is emerging as a lab-tractable model system. Using confocal microscopy and histology, we describe H. miamias reproductive organs, identifying structures previously unknown in acoels. Following a cohort of worms from zygote to adulthood, we quantify the developmental dynamics of their reproductive organs, and find that these organs emerge in a stereotyped sequence as a function of increasing body size. Studying the dynamics of organ growth and de-growth during regeneration and in starvation, we show that reproductive organs follow similar growth rules in these contexts, suggesting that they are regulated by a size-associated program in all growth contexts. Finally, we study egg-laying behavior, finding that H. miamia lay their eggs through their mouths after loading them into their pharynges. Worms lay eggs for multiple months after a single mating, suggesting long-term sperm storage despite lacking a storage organ; we also find that worms can lay viable eggs without mating, indicating a capacity for self-fertilization. Further, we show that worms assess their environment to make decisions about when and where to lay their eggs, and sometimes lay eggs in communal clutches. Together, our work establishes foundational knowledge to enable the experimental study of reproductive anatomy, physiology, and behavior in acoels.

zoology↗

bioRxiv2023

PCP components control anterior and posterior regeneration, with a Prickle homolog impacting muscle organization, in the acoel Hofstenia miamia

Whole-body regeneration requires wound response signals to control patterning programs to enable replacement of structures in their correct locations. While a number of molecular mechanisms underlying anterior-posterior regeneration have been identified, how small fragments of animals first re-establish polarity is less well understood, with non-canonical Wnt signaling recently emerging as a potential regulator. Here, we used the acoel worm Hofstenia miamia, a new research organism capable of robust whole-body regeneration, to assess functions of the components of the Planar Cell Polarity (PCP) pathway in establishing regeneration polarity. We identified homologs of Prickle (pk-1) and Diego (dgo-1) to be required for head and tail regeneration, respectively. RNA-sequencing analysis and experimental corroboration revealed that pk-1 RNAi resulted in diminished expression of early wound response genes as well as of wound-induced expression of the anterior-specific marker fz-7, specifically in tail fragments. In contrast, dgo-1 RNAi impacted wound-induced expression of the posterior-specific marker tf7l2, specifically in head fragments. Furthermore, pk-1 and dgo-1 are enriched in longitudinal muscle, with muscle fibers showing disorganized morphology at anterior-facing wound sites of tail fragments under pk-1 RNAi. These findings suggest that pk-1 and dgo-1 are needed for wound-induced expression of anterior- and posterior-specific genes, and raise the possibility that this action is mediated via the control of muscle fiber orientation. Our work expands the study of PCP genes by revealing their functions in the process of whole-body regeneration in acoels, the sister-group to all other animals with bilateral symmetry, and will enable future studies of PCP components in controlling cellular and tissue-wide regeneration polarity.

developmental biology↗

bioRxiv2022

Acoel single-cell atlas reveals expression dynamics and heterogeneity of a pluripotent stem cell population

Pluripotent adult stem cell populations underlie whole-body regeneration in many distantly related animal lineages. These collectively pluripotent populations of cells share some features across species, such as the expression of piwi and other germline-related genes. Studies of how these cells operate during regeneration are needed in diverse systems to determine how underlying cellular and molecular mechanisms of renewal and differentiation compare. Here, we sought to characterize stem cells and their dynamics in the acoel Hofstenia miamia, a highly regenerative marine worm with a piwi-expressing stem cell population called neoblasts. Transcriptome profiling at single cell resolution revealed cell types shared across postembryonic stages, including stem cells and differentiated cell types such as neural, epidermal, muscle, and digestive cells. Reconstruction of single-cell differentiation trajectories followed by functional studies confirmed that neoblasts are the source of differentiated cells and identified transcription factors needed for the formation of major cell types. Next, analysis of single-cell transcriptomes from regenerating worms showed that both differentiated cells and stem cells dynamically alter gene expression in response to amputation. Further analysis of the stem cells recovered subpopulations of neoblasts, each with specific transcriptional profiles suggesting that the majority of neoblasts are specialized to differentiated lineages, reflecting putatively lineage-primed progenitors. Notably, neoblast subsets in Hofstenia were identifiable consistently across postembryonic stages and also displayed differential expression dynamics in response to wounding. Altogether, these data suggest that whole-body regeneration is accomplished by the coordination of cells with distinct and dynamic transcriptomic profiles through time. Furthermore, the data generated here will enable the study of how this coordination is achieved, enhancing our understanding of pluripotent stem cells and their evolution across metazoans.

developmental biology↗