User:Vellutini

Bruno C. Vellutini is a participant of the EMBO practical course Light sheet microscopy 2014.

I am a PhD student from the Hejnol Group at the Sars International Centre for Marine Molecular Biology in Bergen, Norway. My academic background is based on invertebrate zoology and comparative embryology and I am currently studying the evolution of development and larval body patterns of non-model marine invertebrates, such as bryozoans, brachiopods, and priapulids. I am interested in how SPIM can be used as a platform for visualizing and computing data about the embryonic development.

Background
Spiral cleavage is the remarkably conserved iconic pattern of embryonic cell divisions and developmental fate map characteristic of a major branch of invertebrates, the Spiralia. In a typical spiralian embryo with equal-sized blastomeres the dorsoventral axis is specified by cellular interactions at the 32-cell stage when one vegetal cell contacts the apical cells and becomes an embryonic organizer. The MAPK pathway is the putative signaling cascade regulating this event. MAPK is active exactly in the organizer cell and embryos treated with an inhibitor of the pathway become radialized (i.e., fail to establish the dorsoventral axis).

Even though such key event in spiralian development was described almost forty years ago there is no real-time imaging depicting these cellular interactions. What is really happening at the onset of dorsoventral axis specification in terms of developmental dynamics? Can we further understand the role of MAPK pathway when comparing cellular interactions under normal and experimental conditions? Approaching these questions requires live imaging with a high spatial and temporal resolution, i.e. SPIM ;)

Objectives

 * 1) Describe the interactions between macromeres and micromeres during dorsoventral specification of a spiralian embryo.
 * 2) Compare normal development to the developmental dynamics of embryos treated with an inhibitor of the MAPK pathway.

Organisms


An equally-cleaving annelid or mollusc where the macromere interaction is known to occur would be the ideal animal model. However, I am excited to try these experiments in a different spiralian. Bryozoans are marine colonial invertebrates that have a unique biradial cleavage pattern and stereotypic development, but lost the spiral arrangement of the blastomeres. I am currently tracking the cell lineage of a bryozoan species to solve the fate of the blastomeres and I have found some similarities to the typical spiral cleaving embryos like the origin of larval structures and MAPK activity during embryogenesis. Thus, getting a more comprehensive understanding of bryozoan development with SPIM data can provide insights into the evolution of cleavage patterns in spiralians.

Bryozoan checkpoints


In order to successfully image bryozoan development with lightsheet microscopy some pre-requisites must be accomplished:

1st try

 * 1) Collection of adult colonies in Bergen.
 * 2) Shipping of live colonies from Bergen, Norway to Dresden, Germany.
 * 3) * Shipped: 19/08/2014. Estimated delivery: 20/08/2014.
 * 4) * Last known status: 20 Aug 2014 | 11:54:00 | Dresden | Shipment Delivered In Good Condition. (live status)
 * 5) Passing through customs. CLEARED
 * 6) Arriving alive. NO SURVIVORS. PLASTIC BAG RIPPED OPEN DURING THE JOURNEY.

2nd try

 * 1) Collection of adult colonies in Bergen.
 * 2) Shipping of live colonies from Bergen, Norway to Dresden, Germany.
 * 3) * Shipped: 21/08/2014. Estimated delivery: 22/08/2014.
 * 4) * Last known status: 21 Aug 2014 | 20:34:00 | Stavanger | Shipment In Transit. (live status)
 * 5) Passing through customs.
 * 6) Arriving alive.
 * 7) Adapting to their new home at MPI-CBG.
 * 8) Actually having embryos to share.
 * 9) Walk through sample preparation; to be detailed later.