The Woods Hole image voting posts are some of the most popular posts on the Node (and yes, there will be a new one up VERY soon!). These images are all made by students of the Woods Hole Embryology course, and you still have a chance to be part of the 2013 class!

The application deadline for all Woods Hole summer courses, including this one, has been extended to February 8th. The course itself runs from June 1 to July 14, and is open to graduate students, postdocs, and junior faculty.

Scholarships are available for accepted students, so don’t let money be an issue in your decision to apply.

For more information, see the course website. Good luck! We hope to see some of your images and posts on the Node in the coming year…

(1 votes)

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Hello, my name is Stephanie and I’m a graduate student in Dr. Amy Ralston’s lab at the University of California Santa Cruz.  I just returned from a trip to Dr. Yojiro Yamanaka’s lab at McGill University in Montreal, Quebec.  This trip was funded by the Development Travelling Fellowship from Company of Biologists.  I highly recommend checking it out, receiving this grant was a great, hassle-free experience!

In my time at Dr. Yamanaka’s lab I learned how to synthesize and  inject mRNA constructs into 2 and 8-cell mouse embryos.  I also learned how to live image the embryos as they develop and analyze the data gathered from the imaging.  During my visit I injected GFP and RFP mRNA for easy visualization of my injection success.  I will be using these techniques back at UCSC to study the molecular regulation of the first lineage decision in the mouse embryo.  The molecular mechanisms underlying the first asymmetries and subsequent lineage decisions in the mouse embryo are only beginning to be understood.  I will be using microinjection to over-express a variety of intra-cellular signaling molecules and transcription factors and then assessing the fate of the injected cells.

My time in Montreal was very cold!  Coming from California I’m not adapted to living in subzero temperatures, and most days were below zero (Celsius).  In fact, on the coldest day the high was -22 C, and felt even colder because of the windchill.  It was great to visit French Canada though, very different from other regions in Canada.  McGill University was very international and I met scientists from all over the world who are working there now.  I’m glad to be back in California though, where the high today in Santa Cruz was 15C, well above zero.  I’m also excited to get our injection system up and running and start collecting data.  I included a picture of a 16 cell mouse embryo which I injected at the 8 cell stage.  One cell was injected with H2B conjugated to RFP, marking the DNA, and that cell has subsequently divided.  This was fun and very technically challenging to learn!

(4 votes)

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O-GlcNAc signalling during embryonic stem cell differentiation

My lab is studying the signalling mechanisms governing the onset of differentiation of pluripotent embryonic stem (ES) cells. Work from this and other labs (e.g. 1, 2) has revealed a critical role for autocrine FGF signalling and consequent sustained phosphorylation of the kinase Erk during the differentiation process. Although essential to the differentiation process, Erk activation is not the only signalling event regulating the decision between self-renewal and differentiation of these cells (3). Cellular signalling is most commonly associated with post-translational modification of proteins by addition of a phosphate to serine, threonine and tyrosine residues by the large family of kinase enzymes. However, there are other protein post-translational modifications with increasingly recognised very important roles in protein control. One of these is the addition of β-O-linked N-acetylglucosamine (O-GlcNAc) to serine or threonine residues, first described over 25 years ago. This modification (O-GlcNAcylation) involves the covalent addition of a single sugar to aminoacids via O-glycosidic linkage and occurs with similar time scales, dynamics and stoichiometry as protein phosphorylation.

Compared to the body of work accumulated around the study of phosphorylation, O-GlcNAcylation is much less understood, and relatively little is known about the types of extracellular signals controlling it. However, as these two modifications can occur (mutually exclusively) on the same residue or (in an antagonistic or synergistic fashion) on neighbouring ones, it is easy to see how O-GlcNAcylation can modulate the phosphorylation downstream of a large number of signal transduction cascades (4).

In recent years evidence has been accumulating for a critical role played by O-GlcNAcylation in ES cells, although its precise function(s) and the mechanisms operating are still poorly defined. This project aims to study in detail the role of O-GlcNAcylation on cell signalling, using ES cells as a model system. ES cell differentiation is a complex process, governed by the interaction of multiple signalling pathways (e.g. ERK, Gsk3/Wnt, BMPs etc.) Work in our lab has identified an important and novel role for O-GlcNAc during mouse ESC differentiation, and we have generated preliminary data showing how alteration of O-GlcNAc levels (using a specific inhibitor of the O-GlcNAc hydrolase, GlcNAcstatin, abbreviated GNS) affects cell signalling, gene expression and the self-renewal/differentiation balance. Other labs have recently reported that O-GlcNAc modification of the ESC transcription factors Oct4 and Sox2 controls their function suggesting further mechanisms by which O-GlcNAc profoundly affects cell behaviour (5,6)

This project will build on these preliminary findings and define the mechanisms by which O-GlcNAc affects ES cell function using cell biological, biochemical and proteomic approaches.

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PhD project title:

Non-genetic mechanisms of inheritance influenced by the maternal environment

Project description:

In the last few decades, it has been becoming increasingly clear that organisms can express phenotypes that are inherited in a non-Mendelian fashion. Studies in humans, for example, show that grandfathers who experienced dramatic diet fluctuations as children influenced the incidence of heart disease in their grandchildren. The mechanisms for transgenerational effects, however, are poorly known.

The aim of this project is to study the effect of maternal environment on sex determination of a species of nematode. In this nematode, if the mother smells a specific chemical, she will give rise mostly to hermaphrodites. Otherwise, the mother will produce mostly females. The student will investigate the mechanisms by which an odorant signal can change the epigenetic status of the germline, thereby influencing sex determination. This project will involve RNA sequencing of the germline of mothers that experience odorant stimuli. Mechanistic tests will be performed by genetic manipulation of the nematodes either by mutational analysis or RNA interference.

Key experimental skills involved:

The student will gain experience in analyzing large datasets derived from next-generation RNA sequencing, as well as standard methods in molecular biology and model systems, such as generation of transgenics and RNA interference.

References:

Kaati, G., Bygren, L. O., Pembrey, M. and Sjostrom, M. (2007). Transgenerational response to nutrition, early life circumstances and longevity. Eur J Hum Genet 15, 784-790.

Jablonka, E. (2012). Epigenetic inheritance and plasticity: The responsive germline. Prog Biophys Mol Biol. xxx 1-0 (advanced online).

Chaudhuri, J., Kache, V. and Pires-daSilva, A. (2011). Regulation of sexual plasticity in a nematode that produces males, females, and hermaphrodites. Curr Biol 21, 1548-1551.

Shakes, D. C., Neva, B. J., Huynh, H., Chaudhuri, J. and Pires-daSilva, A. (2011). Asymmetric spermatocyte division as a mechanism for controlling sex ratios. Nat Commun 2, 157.

Contact details for application enquiries:

http://www2.warwick.ac.uk/fac/sci/lifesci/study/pg/research/phd/studentships/#SLS_PhD_Pires

http://www2.warwick.ac.uk/study/postgraduate/apply/

andre.pires@warwick.ac.uk

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Developmental biologists have long relied on the power of observation to understand how embryos develop. In addition, pharmacologic and genetic manipulation of embryos gives us clues as to the mechanisms involved in proper developmental processes. The ability to combine embryo manipulation with observation of embryonic development in real time has been possible for quite some time when using model organisms that develop externally, such as chicks, frogs and zebrafish. However, for those of us that use a mammalian model system, the technology to observe development in real time has lagged way behind. How we long for a way to watch organ systems develop and cell populations migrate and differentiate in the early embryo. While we have many sophisticated genetic tools to study these types of processes, we are limited to looking at simple “snapshots” of time based on when the embryos are dissected and fixed. The task of generating a robust confocal microscopy-based live imaging platform for early mouse embryos was taken on by R’ada Massarwa, a post-doc in the Niswander lab and the culmination of this work was recently published in Development (2013 Jan;140(1):226-36).

During the creation of this live imaging system, she chose to observe the process of neural tube closure, which occurs between days 8.5 and 10.0 of embryonic growth (E8.5-E10.0) in the mouse. Following dissection and experimental setup, the embryos are able to survive up to 16 hours of live imaging. Thus, by performing a series of experiments in which embryos were dissected at increasing somite stages, the entire process of neural tube closure was observed. This type of careful sequential experimentation also showed that the culture and imaging system did not interfere with the proper growth and movement of the tissues. The result: beautiful movies that not only show us how the neural tube develops, but also highlight all the exciting possibilities that this system brings to the study of early mammalian development.

We have been using this system to study neural tube closure, but there are many other tissues and organs that develop during these time periods (E8.5-E10.5) that are amenable to imaging including the heart, face, limbs and neural crest. By using tissue-specific Cre- recombinase reporter strains, the behavior of individual cell types can now be observed in real time in the early mammalian embryo. Also, combining fluorescent reporter strains with genetic knock-out strains and imaging the mutant embryos as the phenotype begins and progresses can provide a much better understanding of how the loss of gene function affects a developmental process. This system also provides access to the embryo itself for pharmacologic and physical manipulation. Overall, the potential for what can be learned using this live embryo imaging system is incredible. We are excited to share this technology with the scientific community and we look forward to seeing how all of you are able to use it to your advantage. Happy imaging!

Massarwa R. & Niswander L. (2012). In toto live imaging of mouse morphogenesis and new insights into neural tube closure, Development, 140 (1) 226-236. DOI: 10.1242/dev.085001

(5 votes)

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Every time a biologist drives pluripotent cells to differentiate into a specialized cell type, patients of all sorts of diseases, disorders, and injuries allow their hope to grow.  A research group recently reported how to drive differentiation of human pluripotent stem cells into medium-sized spiny neurons, the neurons that are some of the first to undergo degeneration in Huntington’s Disease.

hPS (human pluripotent stem) cells have the ability to differentiate into countless specific cell types, and can be either human embryonic stem cells or induced pluripotent cells.  hPS cells can generate various neuronal cell types, so their use in studying neurological diseases and regenerative therapies for such diseases is notable.  Huntington’s disease is an untreatable genetic neurodegenerative disease that typically begins with the degeneration of medium-sized spiny neurons (MSNs), neurons found in the basal ganglia region of the brain.  A recent paper in Development describes how hPS cells can be driven to an MSN fate.  Carri and colleagues began a combinatorial modulation of the pathways involved, beginning with BMP/TGFβ pathway inhibition.  About 20% of the neurons differentiated from hPS cells in these experiments are DARPP-32+/CTIP2+ MSNs also containing dopamine D2 and A2a receptors.  These resulting MSNs showed a firing pattern and neuromodulation identical to mature, authentic MSNs.  Carri and colleagues transplanted the hPS cell-induced neurons into the striatum of acid-lesioned rats, leading to their in vivo survival and differentiation towards an MSN fate.  In the images above, hPS cells were differentiated into neurons that contained DARPP-32 (green), a marker for MSN identity.

For a more general description of this image, see my imaging blog within EuroStemCell, the European stem cell portal.

Carri, A., Onorati, M., Lelos, M., Castiglioni, V., Faedo, A., Menon, R., Camnasio, S., Vuono, R., Spaiardi, P., Talpo, F., Toselli, M., Martino, G., Barker, R., Dunnett, S., Biella, G., & Cattaneo, E. (2012). Developmentally coordinated extrinsic signals drive human pluripotent stem cell differentiation toward authentic DARPP-32+ medium-sized spiny neurons Development, 140 (2), 301-312 DOI: 10.1242/dev.084608

(1 votes)

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We haven’t posted one of these in a while, but there are quite a few things coming up. Remember to also keep an eye on the calendar, and add events there.

Conference and course deadlines in the next few weeks:
January 14 – abstract submission deadline for the annual meeting of the Dutch Society for Developmental Biology
The meeting is January 30 in Utrecht.

January 18 – abstract submission deadline for the joint meeting of the British Societies for Cell and Developmental Biology
March 17-20 Warwick University
Early registration discount ends February 15.

January 31 – abstract submission deadline for the International joint meeting of the German Society for Cell Biology and the German Society for Developmental Biology
March 20-23 Heidelberg
Early registration discount ends February 15

February 1 – application deadline for the Woods Hole Embryology Course
(This is the course that produces the wonderful images that you’ve seen on the Node and on Development covers.)
June 1 – July 14 2013

Other deadlines:
The deadline to apply for the job of Community Manager for the Node is on January 20.

Less urgent, but worth noting:
Registration will soon* open for the 17th International Congress of Developmental Biology. This meeting is only held every four years, and this year it’s in Cancun, so you won’t want to miss this!

(* I first wrote “January 21st”, but had it mixed up with the JSDB meeting )

(1 votes)

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Abcam is a leading web based business supplying research tools to life scientists worldwide. Our head office is in Cambridge (UK), we also have offices in Bristol (UK), Cambridge, Eugene and San Francisco, (USA), Tokyo (Japan), Hong Kong and Hangzhou (China).

A position has become available to develop the product portfolio in the Epigenetics and Nuclear Signalling research area.

You will be part of a larger team responsible for identifying and prioritising targets for antibody production at Abcam. You will be key in ensuring the antibodies produced receive the highest level of validation and data using both internal and external resources. The role will focus on investigating key topics within the field of Epigenetics and Nuclear Siganalling and driving antibody production to meet customer needs. The ideal candidate will be flexible, work well in a team, be comfortable working to deadlines, prioritising different tasks and have good attention to details. Excellent communication skills are essential. You will have contacts within the scientific community and be confident networking to establish new links. Research experience in the field of Epigenetics, Chromatin or Nuclear Signalling is essential.

This position will provide an exciting opportunity for a motivated individual with a PhD or significant research experience who is looking to make the step into a more commercial environment. All relevant training will be provided.

Key Responsibilities:
i) Develop a strategy to identify new targets for antibody production in the field of Epigenetics and Nuclear Signalling. Prioritise production of the most commercially and scientifically valuable antibodies
ii) Post-production validation and characterization of existing antibodies using both internal and external resources.
iii) Networking with the scientific community to identify unmet needs and new product opportunities for Abcam
iv) Providing scientific guidance and input to troubleshoot production or QC issues for key products, working closely with our laboratory and other members of the team
v) Identify current and upcoming scientific topics in order to provide data/information to the Marketing department, including but not limited to, topics for marketing literature, scientific meetings, top selling products.

Our culture is one that empowers individuals, with responsibility given at an early stage. We place great emphasis on knowledge and experience. The working environment is fun and fast-paced, with everybody working together as a team to deliver great service and the best products to our customers. In addition to competitive salaries we can offer an attractive flexible benefits package which includes a profit-share scheme and share options.

To apply, or for more information please follow this link and submit your CV and cover letter: http://hire.jobvite.com/j/?cj=ozd0Wfws&s=The_node

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Here at Development, we’re delighted to welcome François Guillemot to the team of academic editors. François will be replacing Alexandra Joyner, who is soon to step down as our neurodevelopment expert. François heads up the Division of Molecular Neurobiology at the National Institute for Medical Research in London, and his research focuses on the regulation of neurogenesis in the mouse forebrain.

I’d like to take this opportunity to thank Alex for her dedication to and enthusiasm for the journal over the last five years, and to welcome François on board. Given that they used to work together, it should be a seamless transition!

(3 votes)

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A Postdoctoral Fellow position is available in my laboratory.

The overall goal of our laboratory is to understand the cellular and molecular basis of vertebrate organogenesis. Our primary focus is on the elucidation of the mechanisms that govern fate decision and cellular plasticity within the endoderm, for example between pancreas and liver. Toward this aim we perform comparative studies using both amphibian and mammalian model systems, including mouse models and embryonic stem cells.

This position seeks a highly motivated individual with a strong interest in developmental biology. Successful candidate should have a recent Ph.D. or M.D./Ph.D. degree, with strong expertise in one or more of the following areas: genetics, cell and molecular biology, and biochemistry. Prior experience with mouse models and ES cells is a plus. The applicant should be independent and collaborative to be part of a young team and be available for an interview.

Please send your CV, a brief description of your research, and contact information of three references to:

FRANCESCA M. SPAGNOLI, MD PhD

Laboratory of Molecular and Cellular Basis of Embryonic Development
Max Delbrück Center for Molecular Medicine
Robert-Rössle-Str. 10
13125 Berlin
Germany
francesca.spagnoli [a] mdc-berlin.de

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