Summer Nights Nerfin

She favors direct contact with her clients and the search for transactional solutions, especially in family matters.

MAIN FIELDS OF ACTIVITY • Family law • Social insurance law • Lease law • Aliens law • Real estate law • Law of prosecution and bankruptcy • Inheritance law • Labor law • Criminal law.

After an internship in Bernheim, Ming, Gouzer-Waechter & Halpérin, she worked as an associate of François Bolsterli until 1992, then as a partner, until 1995, when she became junior partner of the Study Valticos, Reymond, Mosimann and Veuillet. In 1997, she opened her own studio, in association with Jacques Berta and Enrico Scherrer. In 2003, she joined, with Jacques Berta, the study of Laurence Bory and Emmanuel Ducrest.

The Study, which was enriched by the presence of new partners, namely Mr. Vincent Spira in 2008 and Mr. Emmanuelle Guiguet in 2016, moved on August 1, 2016 to new offices located at 1, place Longemalle.

Here, we show that the zinc-finger transcription factor Nervous fingers 1 (Nerfin-1) is expressed in early-stage medulla neurons and is essential for maintaining their differentiated state. Loss of Nerfin-1 activates Notch signaling in medulla neurons, which promotes neuron dedifferentiation. Inhibition of Notch activity largely caused by tumor-induced depletion and tumorigenesis.

Dedifferentiation caused by Nerfin-1 absence is suppressed by Delta knockdown. (A-B ") Representative nerve-1 159 clones with and without Delta RNAi showing Dpn and Ase staining. Insets show clone regions schematically. Dashed lines outline the clone region. Scale bars: 20 μm. (C) Quantification of the ratio of Dpn + or Ase + cells in clones from A and B (n = 6 for each). Data are mean ± s.e.m .; * P 159 clones, but such upregulation was largely blocked by Notch repression (data not shown). These results suggest that CycE is not a direct target of Nerfin-1 for maintaining medulla neuron differentiation. CycE acts downstream of Notch signaling or it is subsequently upregulated after cell type change.

Nerfin-1 absence results in dedifferentiation of medulla neurons. (A) Time-course experiment of control and nerve-159. Representative NB lineages labeled by GFP are shown. NBs, GMCs and neurons are indicated by asterisks, arrowheads and arrows, respectively. (B) Quantification of the number of GMCs in clones from A. Data are mean ± s.e.m. n.s., not significant. (C-D ') Representative clones of control (C, C') and nerfin-1 159 (D, D '), showing Dpn and Elav staining. Asterisks in C, D 'indicate nonspecific signals of Elav antibody. Arrows indicate Elav - cells. Arrowheads indicate Elav + cells. Insets show clone regions schematically. The temporal expression of Dpn, Mira, Ase and Elav during medulla neuronal differentiation. (F, F ') Ectopic NBs are detected in flip-out clones of Nerfin-1 RNAi. Arrows indicate the clones. Insets show clone regions schematically. (G, H) Nerfin-1 knockdown leads to ectopic NBs in the medulla cortex. Dicer2 (Dcr2) is used to enhance the function of RNAi. Dashed lines represent the edge of medulla neurons. Insets show the medulla cortex schematically. Arrows in H show ectopic NBs among neurons of late stage. (I) Quantification of the number of Dpn + cells according to G, H (n = 12 for each). Data are mean ± s.e.m .; *** P + cells were detected in post-mitotic neurons below the surface (Fig. 4F, F '). Furthermore, elav-Gal4, a pan-neuronal Gal4, was used to express Nerfin-1 RNAi in medulla neurons. However, elav-Gal4 is not strictly expressed in neurons only. Thus, a temperature-sensitive Gal80 protein (Gal80 ts) approach was used to delay the expression. If Nerfin-1 depletion does not function in neurons, those generated before Nerfin-1 RNAi misxpression should remain differentiated and ectopic NBs should be found only in the superficial layer. Interestingly, ectopic NBs have been detected among neurons at different stages (Fig. 4G-I), suggesting that Nerfin-1 is essential for medulla neurons to maintain their differentiation..

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Cellular dedifferentiation is the regression of a cell from a specialized state to a multipotent state and is implicated in cancer. However, the transcriptional network that prevents differentiated cells from reacquiring stem cells is so far unclear. Neuroblasts (NBs), the Drosophila neural stem cells, are a model for the regulation of stem cell self-renewal and differentiation. Here we show that the Drosophila zinc finger transcription factor Nervous fingers 1 (Nerfin-1) locks neurons into differentiation, preventing their reversion into NBs. Following Prospero-dependent neuronal specification in the ganglion mother cell (GMC), a Nerfin-1-specific transcriptional program supports differentiation in the post-mitotic neurons. The loss of Nerfin-1 causes reversion to multipotency and results in tumors in several neural lineages. Both the onset and the rate of neuronal dedifferentiation in nerve-1 mutant lineages are dependent on Myc- and target of rapamycin (Tor) -mediated cellular growth. In addition, Nerfin-1 is required for NB differentiation at the end of neurogenesis. RNA sequencing (RNA-seq) and chromatin immunoprecipitation (ChIP) analysis show that Nerfin-1 administers its function by repression of self-renewing-specific and differentiation-specific activation genes. Our findings support the model of bidirectional interconvertibility between neural stem cells and their post-mitotic progeny and the importance of the Nerfin-1-regulated transcriptional program in neuronal maintenance.

The homeodomain transcription factor has been shown to promote neuronal differentiation (Bello et al 2006, Betschinger et al 2006, Choksi et al 2006). In 17 clones, GMCs failed to give rise to neurons, resulting in tumor clones exclusively of NBs (Fig. 5B-C "'). To compare the transcriptional networks required for neuronal maintenance by Nerfin-1 versus neuronal differentiation promoted by Pros, we conducted a genome-wide transcriptional profile analysis of nerfin-1 159 and 17 mutant mutant clones. GFP-labeled mutant or control cells were isolated through fluorescence-activated cell sorting (FACS) as described in Berger et al. (2012). Total RNA was extracted, sequenced (see Materials and Methods), and compared to control to identify differentially expressed genes. We found a strong correlation between genes that were deregulated in both nerfin-1 159 and 17 clones (Supplemental Figs., S12A-D, S13), with 82% overlap between up-regulated genes in nerfin-1 159 and pros. , few genes were commonly down-regulated in -129 and 17 clones, with only 44% overlap. We have compared our list of genes with NB-and neuronal-specific genes previously identified through transcriptional profiling of FACS-released NBs and neurons (Berger et al., 2012) and found that 25 NB genes were up-regulated and 64 neuronal genes were down-regulated in both pros 17 and nerfin-1 159 mutant clones (data not shown). We have validated a group of these differentially expressed genes by quantitative PCR (qPCR), confirming the up-regulation of the NB-specific genes and the downregulation of the neuronal gene in both mutant tissues (Supplemental Fig. S14). . In addition, type I (Supplemental Fig. S12F-G ") and type II (Supplemental Fig. S5M-N", Q) are cloned. Interestingly, Dpn +, Mira - cells were observed in 1,259 mutant clones, suggesting that Dpn derepression precedes the onset of Mira expression in the process of dedifferentiation (Supplemental Fig. S12G - G ", yellow arrow). Furthermore, we found up-regulation of the GMC-specific target of pox-n (tap) (Bush et al., 1996) and NB-specific gene zipper (zip) in nerfin-1 159 mutant cells. Mammodian encoding of the mammalian protein NeuroD, which has been previously shown to be a target of the Nerfin-1 mammalian INSM1 homolog (Liu et al., 2006). Conversely, we found that the neuronal genes Sox21b, scarecrow (scro), and couch potato (cpo) (Kuzin et al 2005, Southall et al., 2014) were down-regulated in both pros 17 and nerfin-1 159 clones. Nerfin-1 and Pros regulates similar sets of genes, enabling them to inhibit self-renewal and promote differentiation.

nerfin-1 159 ectopic NBs arise from dedifferentiation of neurons. And clone were examined at 24 h (A - B '), 48 h (C - D'), and 72 h (E - F ') after induction and stained with the clone. NB marker Mira (red). The parental NBs, identified by their large size and superficial sections of the brain (0 μm), are marked with white arrows, and ectopic Mira + cells are marked with yellow arrows. (A ", B", C ", D", E ", F") The results are summarized in the schematics. (A - B ") No ectopic NBs have been identified in nerfin-1 159 and control clones at 24 h after induction clone. (C - F ") Ectopic NBs were detected in 1,259 clones at 48 and 72 h after cloning induction in the deepest sections of the clones. (G) Histogram depicting the frequency of ectopic NBs at 24, 48, and 72 h after clone induction in nerfin-1 159 clones (24 h: m = 0, SEM = 0, n = 22, 48 h: m = 1.36, SEM = 0.36, n = 25, 72 h: m = 5.83, SEM = 1.23, n = 23). (H) Histogram depicting the average ectopic NB diameter at 48 and 72 h after clone induction in nerfin-1 159 clones (48 h: m = 5.03, SEM = 0.17, n = 24, 72 h: m = 6.82, SEM = 0.19 , n = 129). (I) Schematic depicting the EdU pulse-chase experiment in control and nerve-1 159 clones. Animals were fed EdU-containing food for 4 hours at 24 hours after cloning induction and then chased with EdU-free food for 24 h. (J - K ") In control clones, no EdU + / Mira + cells were recovered (0%, n = 18). (L-M ") Inner-1 159 clones, EdU + / Mira + ectopic NBs located in deep sections normally occupied by post-mitotic neurons were recovered in 33% of the clones (n = 18, yellow arrow). (J - L ") Parental NBs are marked with white arrows. Bars, 10 μm. (***) P ≤ 0.001. See also Supplemental Figures S8 - S10.

Nerfin-1 promotes differentiation and prevents neuronal reversion into neuroblasts. (A) Type I Express NBs Mira (red) and Myc (orange) and segregates the homeodomain transcription factor Pros (blue) to the GMC. GMCs divide only once to produce two post-mitotic neurons that express Nerfin-1 (green). (B) Upon the loss of Nerfin-1 in neurons, Myc-dependent cellular growth ensures that neurons reach a size after treatment. The reverted cells first switch on stem cell-specific genes such as Mira while maintaining the expression of neuronal genes such as Pros. As these reverted NBs further increase in size, Pros is likely switched off, and the reverted progenitors begin to cycle at the speed of wild-type neuroblasts and undergo asymmetric self-renewal. This expansion of neural progenitors results in proliferative tumorous masses which consists of a mixture of neurons and NBs undergoing various stages of reversion. (C) When ectopically expressed during development, Nerfin-1 promotes premature neuroblast cell cycle exit via size-symmetric cell divisions.

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Born in 1959, Vincent Spira obtained a law degree at the University of Geneva. Admitted to the Geneva Bar in 1983, he worked for Brunschwig, Biaggi, Lévy until 1987, when he opened his own firm, in association with Jean-Charles Sommer and Christian Marti..

In 2008, he joined as a partner the study of Laurence Bory, Emmanuel Ducrest, Jacques Berta and Corinne Nerfin, now Bory & Associés in 2016.

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