Biology subjects

Gerhold, A. R.

Publications and source records attributed to Gerhold, A. R..

5 recordsLinked to original sources

bioRxiv2026

Expanding the C. elegans toolkit with gonad explants

Animal development is a complex process that requires the coordination of a plethora of pathways in space and time. In several species, the availability of tissue explants has provided a simplified context that facilitates mechanistic investigations, particularly into dynamic events. Here, we demonstrate that extruded C. elegans gonads are a viable tissue explant system for this model organism. Using live-cell imaging, we show that C. elegans gonad explants retain many tissue properties that have been documented in vivo, including mitosis, meiosis, apoptosis and gametogenesis. We further show that C. elegans explants are acutely responsive to treatment by the microtubule depolymerizing drug nocodazole. Our work thus reveals C. elegans gonad explants as a new system in which live-cell imaging and acute drug treatment can be combined to decipher the mechanisms governing germline development.

developmental biology↗

bioRxiv2024

The spatiotemporal distribution of LIN-5/NuMA regulates spindle orientation and tissue organization in the C. elegans germ line

Mitotic spindle orientation sets the cell division plane and is thus critical for maintaining tissue organization. The C. elegans gonad is tube-shaped, with germ cells forming a circumferential monolayer around a shared inner core of cytoplasm called the rachis. Each germ cell is connected to the rachis via a stable cytoplasmic bridge, polarizing germ cells along their rachis-basal axis. How this tissue organization is maintained during development is unclear, as germ cells lack the canonical cell-cell junctions that, in other tissue types, ensure proper spindle orientation. Here we use live-cell imaging of C. elegans germ cells, both in situ and in gonad explants, to show that the microtubule force generator dynein and its conserved regulator LIN-5/NuMA regulate spindle orientation in C. elegans germ cells and are required for germline tissue organization. We uncover a cyclic, polarized pattern of LIN-5/NuMA cortical localization that predicts centriole/centrosome positioning throughout the cell cycle, providing a means to align spindle orientation with the tissue plane. This work reveals a new mechanism by which oriented cell division can be achieved to maintain tissue organization during animal development.

cell biology↗

bioRxiv2024

Non-autonomous insulin signaling regulates the duration of mitosis in C. elegans germline stem and progenitor cells

Stem and progenitor cell mitosis is essential for tissue development and homeostasis. How these cells ensure proper chromosome segregation, and thereby maintain mitotic fidelity, in the complex physiological environment of a living animal is poorly understood. Here we use in situ live-cell imaging of C. elegans germline stem and progenitor cells (GSPCs) to ask how the signaling environment influences stem and progenitor cell mitosis in vivo. Through a candidate screen we identify a new role for the insulin/IGF receptor (IGFR), daf-2, during GSPC mitosis. Mitosis is delayed in daf-2/IGFR mutants, and these delays require canonical, DAF-2/IGFR to DAF-16/FoxO insulin signaling, here acting cell non-autonomously from the soma. Interestingly, mitotic delays in daf-2/IGFR mutants depend on the spindle assembly checkpoint but are not accompanied by a loss of mitotic fidelity. Correspondingly, we show that caloric restriction, which delays GSPC mitosis and compromises mitotic fidelity, does not act via the canonical insulin signaling pathway, and instead requires AMP-activated kinase (AMPK). Together this work demonstrates that GSPC mitosis is influenced by at least two genetically separable signaling pathways and highlights the importance of signaling networks for proper stem and progenitor cell mitosis in vivo. Author SummaryStem and progenitor cells drive tissue development and sustain adult tissue turnover by producing new daughter cells via cell division, the success of which depends on proper chromosome segregation during mitosis. Stem and progenitor cells perform mitosis in the complex environment of a living animal, yet relatively little is known about how events during mitosis are influenced by this in vivo context. In particular, whether signaling pathways that coordinate other aspects of stem and progenitor cell behavior with animal physiology also play a role during mitosis is poorly understood. Here we took advantage of the germline stem and progenitor cells of the model nematode C. elegans to address this question. Through live-cell imaging of germline stem and progenitor cell mitosis, we uncover a new role for the insulin signaling pathway. We find that reducing insulin signaling delays germline stem and progenitor cell mitosis, but, surprisingly, these delays are not accompanied by a loss of mitotic fidelity. In addition, we find that reducing insulin signaling in somatic tissues is sufficient to delay germline stem and progenitor mitosis, indicating that the pathway acts non-autonomously. Finally, while insulin signaling is known to link cell division with nutritional status in many species, we found that it did not mediate the effects of caloric restriction on germline stem and progenitor cell mitosis. Instead, caloric restriction acts via the conserved energy-sensing regulator AMPK. These results uncover new regulators of germline stem and progenitor cell mitosis and emphasize the importance of signaling pathways for proper stem and progenitor cell mitosis in vivo.

developmental biology↗

bioRxiv2020

CentTracker: a trainable, machine learning-based tool for large-scale analyses of C. elegans germline stem cell mitosis

Investigating the complex interactions between stem cells and their native environment requires an efficient means to image them in situ. Caenorhabditis elegans germline stem cells (GSCs) are distinctly accessible for intravital imaging; however, long-term image acquisition and analysis of dividing GSCs can be technically challenging. Here we present a systematic investigation into the technical factors impacting GSC physiology during live imaging and provide an optimized method for monitoring GSC mitosis under minimally disruptive conditions. We describe CentTracker, an automated and generalizable image analysis tool that uses machine learning to pair mitotic centrosomes and which can extract a variety of mitotic parameters rapidly from large-scale datasets. We employ CentTracker to assess a range of mitotic features in GSCs and show that subpopulations with distinct mitotic profiles are unlikely to exist within the stem cell pool. We further find evidence for spatial clustering of GSC mitoses within the germline tissue and for biases in mitotic spindle orientation relative to the germlines distal-proximal axis, and thus the niche. The technical and analytical tools provided herein pave the way for large-scale screening studies of multiple mitotic processes in GSCs dividing in situ, in an intact tissue, in a living animal, under seemingly physiological conditions.

cell biology↗

bioRxiv2020

Widely used mutants of eiger, encoding the Drosophila Tumor Necrosis factor, carry additional mutations in the NimrodC1 phagocytosis receptor

The process of apoptosis in epithelia involves activation of caspases, delamination of cells, and degradation of cellular components. Corpses and cellular debris are then rapidly cleared from the tissue by phagocytic blood cells. In studies of the Drosophila TNF, Eiger (Egr) and cell death in wing imaginal discs, the epithelial primordia of fly wings, we noticed that dying cells persisted longer in egr3 mutant wing discs than in wild type discs, raising the possibility that their phagocytic engulfment by hemocytes was impaired. Further investigation revealed that lymph glands and circulating hemocytes from egr3 mutant larvae were completely devoid of NimC1 staining, a marker of phagocytic hemocytes. Genome sequencing uncovered mutations in the NimC1 coding region that are predicted to truncate the NimC1 protein before its transmembrane domain, and provide an explanation for the lack of NimC staining. The work that we report here demonstrates the presence of these NimC1 mutations in the widely used egr3 mutant, its sister allele, egr1, and its parental strain, Regg1GS9830. As the egr3 and egr1 alleles have been used in numerous studies of immunity and cell death, it may be advisable to re-evaluate their associated phenotypes.

genetics↗