Light Microscopy of Echinoderm Embryos

Strickland, Laila; Dassow, George von; Ellenberg, Jan; Foe, Victoria E.; Lénárt, Péter; Burgess, David R.

doi:10.1016/s0091-679x(04)74016-9

Cited by 43 publications

(38 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To visualise polymerised actin and microtubules, cells were fixed using a previously described protocol for the preservation of cytoskeletal elements. 42,43 To visualise g-tubulin staining, cells were fixed using a methanol fixation protocol, as previously described. 44 Cells were stained with DAPI to visualise nuclei.…”

Section: Methodsmentioning

confidence: 99%

Mouse granzyme A induces a novel death with writhing morphology that is mechanistically distinct from granzyme B-induced apoptosis

et al. 2013

View full text Add to dashboard Cite

Human and mouse granzyme (Gzm)B both induce target cell apoptosis in concert with pore-forming perforin (Pfp); however the mechanisms by which other Gzms induce non-apoptotic death remain controversial and poorly characterised. We used timelapse microscopy to document, quantitatively and in real time, the death of target cells exposed to primary natural killer (NK) cells from mice deficient in key Gzms. We found that in the vast majority of cases, NK cells from wild-type mice induced classic apoptosis. However, NK cells from syngeneic Gzm B-deficient mice induced a novel form of cell death characterised by slower kinetics and a pronounced, writhing, 'worm-like' morphology. Dying cells initially contracted but did not undergo membrane blebbing, and annexin-V staining was delayed until the onset of secondary necrosis. As it is different from any cell death process previously reported, we tentatively termed this cell death 'athetosis'. Two independent lines of evidence showed this alternate form of death was due to Gzm A: first, cell death was revealed in the absence of Gzm B, but was completely lost when the NK cells were deficient in both Gzm A and B; second, the athetotic morphology was precisely reproduced when recombinant mouse Gzm A was delivered by an otherwise innocuous dose of recombinant Pfp. Gzm A-mediated athetosis did not require caspase activation, early mitochondrial disruption or generation of reactive oxygen species, but did require an intact actin cytoskeleton and was abolished by latrunculin B and mycalolide B. This work defines an authentic role for mouse Gzm A in granule-induced cell death by cytotoxic lymphocytes.

show abstract

Section: Methodsmentioning

confidence: 99%

Mouse granzyme A induces a novel death with writhing morphology that is mechanistically distinct from granzyme B-induced apoptosis

et al. 2013

View full text Add to dashboard Cite

show abstract

“…Oocytes were settled onto protaminecoated coverslips for two minutes. Coverslips were then were briefly immersed in an isotonic glycerol buffer 44 before placing in ice-cold fix (40 mM PIPES, 10 mM EGTA, 5 mM MgSO 4 , 1M glycerol, 1% Triton X-100, 3.7% formaldehyde, pH 6.9) for an hour. Sodium fluoride (to ~10 mM) was added to the fixative just before use to act as a phosphatase inhibitor.…”

Section: Methodsmentioning

confidence: 99%

Aurora B Kinase Maintains Chromatin Organization During the MI to MII Transition in Surf Clam Oocytes

George¹,

Johnston²,

Shuster³

2006

Cell Cycle

View full text Add to dashboard Cite

“…To estimate the transition of total cell numbers during the early stages of development in P. baculosa embryos, the number of nuclei per embryo were counted after staining with propidium iodide (Nacalai Tesque), as described previously (Stickland et al, 2004) (supplementary material Table S1). …”

Section: Animals and Embryosmentioning

confidence: 99%

Larval mesenchyme cell specification in the primitive echinoid occurs independently of the double-negative gate

et al. 2014

View full text Add to dashboard Cite

Echinoids (sea urchins) are divided into two major groups -cidaroids (a 'primitive' group) and euechinoids (a 'derived' group). The cidaroids are a promising model species for understanding the ancestral developmental mechanisms in echinoids, but little is known about the molecular mechanisms of cidaroid development. In euechinoids, skeletogenic mesenchyme cell specification is regulated by the double-negative gate (DNG), in which hesC represses the transcription of the downstream mesenchyme specification genes (alx1, tbr and ets1), thereby defining the prospective mesenchyme region. To estimate the ancestral mechanism of larval mesenchyme cell specification in echinoids, the expression patterns and roles of mesenchyme specification genes in the cidaroid Prionocidaris baculosa were examined. The present study reveals that the expression pattern and function of hesC in P. baculosa were inconsistent with the DNG model, suggesting that the euechinoidtype DNG is not utilized during cidaroid mesenchyme specification. In contrast with hesC, the expression patterns and functions of alx1, tbr and ets1 were similar between P. baculosa and euechinoids. Based on these results, we propose that the roles of alx1, tbr and ets1 in mesenchyme specification were established in the common ancestor of echinoids, and that the DNG system was acquired in the euechinoid lineage after divergence from the cidaroid ancestor. The evolutionary timing of the establishment of the DNG suggests that the DNG was originally related to micromere and/or primary mesenchyme cell formation but not to skeletogenic cell differentiation.

show abstract

Light Microscopy of Echinoderm Embryos

Cited by 43 publications

References 30 publications

Mouse granzyme A induces a novel death with writhing morphology that is mechanistically distinct from granzyme B-induced apoptosis

Mouse granzyme A induces a novel death with writhing morphology that is mechanistically distinct from granzyme B-induced apoptosis

Aurora B Kinase Maintains Chromatin Organization During the MI to MII Transition in Surf Clam Oocytes

Larval mesenchyme cell specification in the primitive echinoid occurs independently of the double-negative gate

Contact Info

Product

Resources

About