Spinning Out Greentech Start-ups.

The Paris Agreement has made combating climate change a priority and has incentivised innovationfor the greentech industry. Higher education institution[s] (HEI[s]) play an important role in fueling this innovation by developing disruptive technologies and support the creation of start-up companies that take the risk to bring these technologies to the market. The successful spinning out of such start-ups relies on the HEIs nurturing anecosystem with multiple actors as well as internal mechanisms to transfer such technologies to the start-up. HEIs have dedicated offices involved in technology transfer (TTO) that provide an interface between all the different actors (inventors, founders, start-up, investors, etc.). The TTO of an HEI also plays a central role in the technology transfer by providing the licence agreements granting start-ups the rights to use a technology developed by the HEI. This review summarises the role of a TTO and the practice of making the licensing process as transparent as possible for start-up founders and investors.

Beta- and gamma-cytoplasmic actins are required for meiosis in mouse oocytes.

In mammals, female meiosis consists of two asymmetric cell divisions, which generate a large haploid oocyte and two small polar bodies. Asymmetric partitioning of the cytoplasm results from migration of the meiotic spindle toward the cortex and requires actin filaments. However, the subcellular localization and the role of the existing two cytoplasmic actin (CYA) isoforms, beta and gamma, have not been characterized. We show that beta- and gamma-CYA are differentially distributed in the maturing oocyte from late metaphase I as well as in preimplantation embryos. Gamma-CYA is preferentially enriched in oocyte cortices and is absent from all cell-cell contact areas from metaphase II until the blastocyst stage. Beta-CYA is enriched in contractile structures, at cytokinesis, at cell-cell contacts, and around the forming blastocoel. Alteration of beta- or gamma-CYA function by isoform-specific antibody microinjection suggests that gamma-CYA holds a major and specific role in the establishment and/or maintenance of asymmetry in meiosis I and in the maintenance of overall cortical integrity. In contrast, beta- and gamma-CYA, together, appear to participate in the formation and the cortical anchorage of the second meiotic spindle in waiting for fertilization. Finally, differences in gamma-CYA expression are amongst the earliest markers of cell fate determination in development.

Dcp1-bodies in mouse oocytes.

Processing bodies (P-bodies) are cytoplasmic granules involved in the storage and degradation of mRNAs. In somatic cells, their formation involves miRNA-mediated mRNA silencing. Many P-body protein components are also found in germ cell granules, such as in mammalian spermatocytes. In fully grown mammalian oocytes, where changes in gene expression depend entirely on translational control, RNA granules have not as yet been characterized. Here we show the presence of P-body-like foci in mouse oocytes, as revealed by the presence of Dcp1a and the colocalization of RNA-associated protein 55 (RAP55) and the DEAD box RNA helicase Rck/p54, two proteins associated with P-bodies and translational control. These P-body-like structures have been called Dcp1-bodies and in meiotically arrested primary oocytes, two types can be distinguished based on their size. They also have different protein partners and sensitivities to the depletion of endogenous siRNA/miRNA and translational inhibitors. However, both type progressively disappear during in vitro meiotic maturation and are virtually absent in metaphase II-arrested secondary oocytes. Moreover, this disassembly of hDcp1a-bodies is concomitant with the posttranslational modification of EGFP-hDcp1a.

Fgf2 is expressed in human and murine embryonic choroid plexus and affects choroid plexus epithelial cell behaviour.

BACKGROUND: Although fibroblast growth factor (Fgf) signalling plays crucial roles in several developing and mature tissues, little information is currently available on expression of Fgf2 during early choroid plexus development and whether Fgf2 directly affects the behaviour of the choroid plexus epithelium (CPe). The purpose of this study was to investigate expression of Fgf2 in rodent and human developing CPe and possible function of Fgf2, using in vitro models. The application of Fgf2 to brain in vivo can affect the whole tissue, making it difficult to assess specific responses of the CPe. METHODS: Expression of Fgf2 was studied by immunohistochemistry in rodent and human embryonic choroid plexus. Effects of Fgf2 on growth, secretion, aggregation and gene expression was investigated using rodent CPe vesicles, a three-dimensional polarized culture model that closely mimics CPe properties in vivo, and rodent CPe monolayer cultures. RESULTS: Fgf2 was present early in development of the choroid plexus both in mouse and human, suggesting the importance of this ligand in Fgf signalling in the developing choroid plexus. Parallel analysis of Fgf2 expression and cell proliferation during CP development suggests that Fgf2 is not involved in CPe proliferation in vivo. Consistent with this observation is the failure of Fgf2 to increase proliferation in the tri-dimensional vesicle culture model. The CPe however, can respond to Fgf2 treatment, as the diameter of CPe vesicles is significantly increased by treatment with this growth factor. We show that this is due to an increase in cell aggregation during vesicle formation rather than increased secretion into the vesicle lumen. Finally, Fgf2 regulates expression of the CPe-associated transcription factors, Foxj1 and E2f5, whereas transthyretin, a marker of secretory activity, is not affected by Fgf2 treatment. CONCLUSION: Fgf2 expression early in the development of both human and rodent choroid plexus, and its ability to modulate behaviour and gene expression in CPe, supports the view that Fgf signalling plays a role in the maintenance of integrity and function of this specialized epithelium, and that this role is conserved between rodents and humans.

Growth of choroid plexus epithelium vesicles in vitro depends on secretory activity.

Although a number of models have been used to study choroid plexus epithelium (CPe) function, analysis in physiological conditions of this polarised epithelium which produces the majority of the cerebrospinal fluid (CSF) and is one of the key barriers between blood and CSF in the brain remains challenging. As CPe cells form polarised CPe vesicles when cultured in Matrigel, we have assessed their behaviour and potential use for pharmacological studies. Like CPe cells in vivo, CPe vesicles express transthyretin, E2f5, Fox-j1 and p73, and contain tight junctions, as indicated by ZO-1 expression and electron microscopy analysis. Time-lapse microscopy shows that CPe cells plated in Matrigel are highly migratory and rapidly form homotypic cell aggregates, which then reorganise to form vesicles whose size increases linearly overtime. Neither aggregate nor vesicle size is affected by AraC treatment, though this inhibitor significantly reduces proliferation in CPe monolayers. Increase in size of vesicles, which have reached a growth plateau is observed following addition of fluorescently-labelled CPe cells, which become incorporated into the vesicle walls. Significantly, treatment with secretion inhibitors blocks vesicle formation and their expansion. These results show that secretion, rather than cell division, controls vesicle growth, consistent with low levels of proliferation and thinning of the CPe observed both in growing vesicles and during CPe development. Therefore, changes in vesicle size can be used to evaluate the effect of putative molecules involved in the regulation of secretion.

Changes in E2F5 intracellular localization in mouse and human choroid plexus epithelium with development.

The choroid plexus epithelium (CPe) is a specialized epithelium involved primarily in the production of cerebrospoinal fluid (CSF) which is important for maintaining an optimal homeostatic environment for the brain. Although, the physiology of the CPe is fairly well understood, its development has not been thoroughly studied. It has been recently shown that mice lacking functional transcription factors, E2F5, foxJ1 or p73, develop non-obstructive hydrocephalus likely due to CPe dysfunction. We have further studied their expression in the mouse and human developing CPe, focusing particularly on E2F5. We show here that in the mouse E2F5, foxJ1 and p73 transcripts are detectable as soon as the choroid plexuses form. E2F5 protein is also detected as soon as the choroid plexuses are morphologically apparent both in mouse and human, suggesting that its expression is regulated at the transcriptional level. E2F5 protein is down-regulated late in embryogenesis and this coincides with a change in its intracellular localization, from predominantly nuclear to cytoplasmic. The pattern of expression and intracellular localization of E2F5 in vivo does not appear to correlate with that of proliferating CPe cells, as indicated by protein cell nuclear antigen (PCNA) staining, but rather with their maturation, as changes in E2F5 localization from the nucleus to the cytoplasm parallel the morphological change from pseudostratified to cuboidal epithelium.