Increasing Diversity in Developmental Biology.

The demographic profile of the scientific and biomedical workforce in the United States does not reflect the population at large (https://ncses.nsf.gov/pubs/nsf21321/data-tables; www.census.gov), raising concerns that there will be too few trained researchers in the future, the scope of research interests will not be broad enough, gaps in equity and social justice will continue to increase, and the safeguards to the integrity of the scientific enterprise could be jeopardized. To diversify the pool of scientists, the Society for Developmental Biology (SDB) developed the Choose Development! Program-a two-summer immersion for undergraduate students belonging to underrepresented (UR) populations in STEM to join the research laboratory of an established SDB member. This research-intensive experience was augmented by a multi-tier mentoring plan for each student, society-wide recognition, professional development activities and networking at national meetings. The strengths of the Choose Development! Program were leveraged to expand inclusion and outreach at the Society's leadership level, the Board of Directors (BOD), which then led to significant changes that impacted the SDB community. The cumulative outcomes of the Choose Development! Program provides evidence that community-based, long-term advocacy, and mentoring of young UR scientists is successful in retaining UR students in scientific career paths and making a scientific society more inclusive.

Linfoma pleural asociado a empiema crónico / Pleural Lymphoma Associated With Chronic Empyema

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miR-206 is required for changes in cell adhesion that drive muscle cell morphogenesis in Xenopus laevis.

MicroRNAs (miRNAs) are highly conserved small non-coding RNA molecules that post-transcriptionally regulate gene expression in multicellular organisms. Within the set of muscle-specific miRNAs, miR-206 expression is largely restricted to skeletal muscle and is found exclusively within the bony fish lineage. Although many studies have implicated miR-206 in muscle maintenance and disease, its role in skeletal muscle development remains largely unknown. Here, we examine the role of miR-206 during Xenopus laevis somitogenesis. In Xenopus laevis, miR-206 expression coincides with the onset of somitogenesis. We show that both knockdown and over-expression of miR-206 result in abnormal somite formation affecting muscle cell rotation, attachment, and elongation. In particular, our data suggests that miR-206 regulates changes in cell adhesion that affect the ability of newly formed somites to adhere to the notochord as well as to the intersomitic boundaries. Additionally, we show that ß-dystroglycan and F-actin expression levels are significantly reduced, suggesting that knockdown of miR-206 levels affects cellular mechanics necessary for cell shape changes and attachments that are required for proper muscle formation.

Enabling full representation in science: the San Francisco BUILD project's agents of change affirm science skills, belonging and community.

BACKGROUND: The underrepresentation of minority students in the sciences constrains innovation and productivity in the U.S. The SF BUILD project mission is to remove barriers to diversity by taking a "fix the institution" approach rather than a "fix the student" one. SF BUILD is transforming education, research, training, and mentoring at San Francisco State University, a premiere public university that primarily serves undergraduates and ethnic minority students. It boasts a large number of faculty members from underrepresented groups (URGs), including many of the project leaders. These leaders collaborate with faculty at the University of California San Francisco (UCSF), a world-class medical research institution, to implement SF BUILD. KEY HIGHLIGHTS: Together, the campus partners are committed to creating intellectually safe and affirming environments grounded in the Signaling Affirmation for Equity (SAFE) model, which is based on robust psychosocial evidence on stereotype threat and its consequences. The SAFE model dictates a multilevel approach to increasing intent to pursue a biomedical career, persistence in STEM fields, and productivity (e.g. publications, presentations, and grants) by implementing transformative activities at the institutional, faculty, and student levels. These activities (1) increase knowledge of the stereotype threat phenomenon; (2) affirm communal and altruistic goals of students and faculty to "give back" to their communities in classrooms and research activities; and (3) establish communities of students, faculty and administrators as "agents of change." Agents of change are persons committed to establishing and maintaining SAFE environments. In this way, SF BUILD advances the national capacity to address biomedical questions relevant to communities of color by enabling full representation in science. IMPLICATIONS: This chapter describes the theoretical and historical context that drive the activities, research and evaluation of the SF BUILD project, and highlights attributes that other institutions can use for institutional change. While this paper is grounded in psychosocial theory, it also provides practical solutions for broadening participation.

Making muscle: Morphogenetic movements and molecular mechanisms of myogenesis in Xenopus laevis.

Xenopus laevis offers unprecedented access to the intricacies of muscle development. The large, robust embryos make it ideal for manipulations at both the tissue and molecular level. In particular, this model system can be used to fate map early muscle progenitors, visualize cell behaviors associated with somitogenesis, and examine the role of signaling pathways that underlie induction, specification, and differentiation of muscle. Several characteristics that are unique to X. laevis include myogenic waves with distinct gene expression profiles and the late formation of dermomyotome and sclerotome. Furthermore, myogenesis in the metamorphosing frog is biphasic, facilitating regeneration studies. In this review, we describe the morphogenetic movements that shape the somites and discuss signaling and transcriptional regulation during muscle development and regeneration. With recent advances in gene editing tools, X. laevis remains a premier model organism for dissecting the complex mechanisms underlying the specification, cell behaviors, and formation of the musculature system.

The Role of Sdf-1α signaling in Xenopus laevis somite morphogenesis.

BACKGROUND: Stromal derived factor-1α (sdf-1α), a chemoattractant chemokine, plays a major role in tumor growth, angiogenesis, metastasis, and in embryogenesis. The sdf-1α signaling pathway has also been shown to be important for somite rotation in zebrafish (Hollway et al., 2007). Given the known similarities and differences between zebrafish and Xenopus laevis somitogenesis, we sought to determine whether the role of sdf-1α is conserved in Xenopus laevis. RESULTS: Using a morpholino approach, we demonstrate that knockdown of sdf-1α or its receptor, cxcr4, leads to a significant disruption in somite rotation and myotome alignment. We further show that depletion of sdf-1α or cxcr4 leads to the near absence of ß-dystroglycan and laminin expression at the intersomitic boundaries. Finally, knockdown of sdf-1α decreases the level of activated RhoA, a small GTPase known to regulate cell shape and movement. CONCLUSION: Our results show that sdf-1α signaling regulates somite cell migration, rotation, and myotome alignment by directly or indirectly regulating dystroglycan expression and RhoA activation. These findings support the conservation of sdf-1α signaling in vertebrate somite morphogenesis; however, the precise mechanism by which this signaling pathway influences somite morphogenesis is different between the fish and the frog.

Temporal and spatial patterning of axial myotome fibers in Xenopus laevis.

Somites give rise to the vertebral column and segmented musculature of adult vertebrates. The cell movements that position cells within somites along the anteroposterior and dorsoventral axes are not well understood. Using a fate mapping approach, we show that at the onset of Xenopus laevis gastrulation, mesoderm cells undergo distinct cell movements to form myotome fibers positioned in discrete locations within somites and along the anteroposterior axis. We show that the distribution of presomitic cells along the anteroposterior axis is influenced by convergent and extension movements of the notochord. Heterochronic and heterotopic transplantations between presomitic gastrula and early tail bud stages show that these cells are interchangeable and can form myotome fibers in locations determined by the host embryo. However, additional transplantation experiments revealed differences in the competency of presomitic cells to form myotome fibers, suggesting that maturation within the tail bud presomitic mesoderm is required for myotome fiber differentiation.

Embryonic cells depleted of beta-catenin remain competent to differentiate into dorsal mesodermal derivatives.

Disruption of axis specification leads to defects in dorsal tissue patterning and cell movements. Here, we examine how beta-catenin coordinately affects gastrulation movements and dorsal mesoderm differentiation. The reduction of beta-catenin protein levels by morpholino oligonucleotides complementary to beta-catenin mRNA causes a disruption in gastrulation movements. Time-lapse imaging of beta-catenin morphants during gastrulation reveals that involution occurs simultaneously around the blastopore in the absence of convergent extension cell movements. Transplantation experiments show that morphant cells grafted from the marginal zone into wild-type hosts differentiate into notochord and muscle. However, wild-type mesoderm cells grafted to the marginal zone of beta-catenin morphants do not form dorsal tissues. These data argue that beta-catenin is not required for the initial establishment of dorsal mesoderm cell competency, but it is required for the maintenance of that competency. We propose that tissue interactions that occur during convergent extension movements are necessary for maintaining dorsal tissue competency.

Control of morphogenetic cell movements in the early zebrafish myotome.

As the vertebrate myotome is generated, myogenic precursor cells undergo extensive and coordinated movements as they differentiate into properly positioned embryonic muscle fibers. In the zebrafish, the "adaxial" cells adjacent to the notochord are the first muscle precursors to be specified. After initially differentiating into slow-twitch myosin-expressing muscle fibers, these cells have been shown to undergo a remarkable radial migration through the lateral somite, to populate the superficial layer of slow-twitch muscle of the mature myotome. Here we characterize an earlier set of adaxial cell behaviors; the transition from a roughly 4x5 array of cuboidal cells to a 1x20 stack of elongated cells, prior to the migration event. We find that adaxial cells display a highly stereotypical series of behaviors as they undergo this rearrangement. Furthermore, we show that the actin regulatory molecule, Cap1, is specifically expressed in adaxial cells and is required for the progression of these behaviors. The requirement of Cap1 for a cellular apical constriction step is reminiscent of similar requirements of Cap during apical constriction in Drosophila development, suggesting a conservation of gene function for a cell biological event critical to many developmental processes.

Cell behaviors associated with somite segmentation and rotation in Xenopus laevis.

During vertebrate development the formation of somites is a critical step, as these structures will give rise to the vertebrae, muscle, and dermis. In Xenopus laevis, somitogenesis consists of the partitioning of the presomitic mesoderm into somites, which undergo a 90-degree rotation to become aligned parallel to the notochord. Using a membrane-targeted green fluorescent protein to visualize cell outlines, we examined the individual cell shape changes occurring during somitogenesis. We show that this process is the result of specific, coordinated cell behaviors beginning with the mediolateral elongation of cells in the anterior presomitic mesoderm and then the subsequent bending of these elongated cells to become oriented parallel with the notochord. By labeling a clonal population of paraxial mesoderm cells, we show that cells bend around their dorsoventral axis. Moreover, this cell bending correlates with an increase in the number of filopodial protrusions, which appear to be posteriorly directed toward the newly formed segmental boundary. By examining the formation of somites at various positions along the anteroposterior axis, we show that the general sequence of cell behaviors is the same; however, somite rotation in anterior somites is slower than in posterior somites. Lastly, this coordinated set of cell behaviors occurs in a dorsal-to-ventral progression within each somite such that cells in the dorsal aspect of the somite become aligned along the anteroposterior axis before cells in other regions of the same somite. Together, our data further define how these cell behaviors are temporally and spatially coordinated during somite segmentation and rotation.

Muscle specification in the Xenopus laevis gastrula-stage embryo.

Recent fate maps of the Xenopus laevis gastrula show that mesodermal tissue surrounding the blastopore gives rise to muscle (Keller [1991] Methods Cell Biol 36:61-113; Lane and Smith [1999] Development 126:423-434). In a significant deviation from earlier data, the new maps demonstrate that cells in the ventral half of the gastrula are precursors to a significant portion of trunk somites. However, these posterior somites are not formed until tadpole stages (stages 38-44). We therefore set out to determine the timing of muscle specification within the ventral half of the gastrula. Our approach was to generate a series of tissue explants from gastrula-stage embryos and then culture them to either stage 28 (tailbud) or stage 44 (tadpole). At each endpoint, the presence of muscle in explants was assessed with a muscle-specific antibody. Interestingly, we found that muscle tissue is detected in ventral explants. However, these explants must be cultured to the tadpole stage. This is perhaps not unexpected, as this is the point at which this tissue normally gives rise to muscle. We further show that muscle specification of the involuting marginal zone does not change over the course of gastrulation. Together, these results suggest that dorsalizing signals emanating from the midline during gastrulation are not necessary for muscle specification of the ventral half of the involuting marginal zone.

Signals that instruct somite and myotome formation persist in Xenopus laevis early tailbud stage embryos.

Somite formation is a lengthy process that begins at gastrulation and continues through tailbud stages to form approximately 50 pairs of somites in the frog, Xenopus laevis. In Xenopus, the somite primarily gives rise to myotome. We sought to determine whether the formation of somites and myotome requires a transient signal active during gastrulation or a constitutive signal active throughout development to instruct dorsal mesodermal cells to form the posterior somites. Previous work from our lab revealed that cells from the neural ectoderm are capable of responding to mesoderm-inducing signals [Domingo and Keller: Dev Biol 2000;225:226-240]. Thus, to test for the presence of somite-inducing signals, we performed a series of grafting experiments in which we used gastrula cells from the anterior neural ectoderm (ANE). Fluorescently labeled ANE cells were grafted to the posterior paraxial mesoderm of progressively older host embryos between stages 11 (mid gastrula) and 23 (early tailbud). Our results showed that signals within the paraxial mesoderm can instruct prospective ANE cells, which normally give rise to head structures, to instead differentiate into myotome cells. We found that the grafted cells adopted the local paraxial mesoderm cell behaviors, which consists of mediolateral intercalation, segmentation, somite cell rotation, and differentiation to myotome. In addition, we show that the grafted ANE cells that adopt a myotome morphology also express the muscle-specific marker, 12/101. Through a series of heterochronic grafts, we determined that the duration of somite-inducing signals extends from the early gastrula (stage 11) through the early tailbud (stage 23) stage embryos. These results demonstrate that somite induction is not a transient event that occurs during gastrulation, but that it is instead a continuous event that can occur as new somites are added to the posterior axis.