Direct Conversion of
            <i>C. elegans</i>
            Germ Cells into Specific Neuron Types

Tursun, Baris; Patel, Tulsi; Kratsios, Paschalis; Hobert, Oliver

doi:10.1126/science.1199082

Cited by 215 publications

(288 citation statements)

References 29 publications

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“…The development of different neuronal subtypes can be induced in the C. elegans germline by simultaneous removal of certain chromatin factors and forced expression of master neuronal transcription factors (Tursun et al 2011;Patel et al 2012), highlighting the plasticity of the germline. Notably, other genetic perturbations have been shown to cause C. elegans germ cells to express neuronal and muscle markers, without the need for driving transcription factors.…”

Section: Discussionmentioning

confidence: 99%

Germ Granules Prevent Accumulation of Somatic Transcripts in the AdultCaenorhabditis elegansGermline

et al. 2017

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The germ cells of multicellular organisms protect their developmental potential through specialized mechanisms. A shared feature of germ cells from worms to humans is the presence of nonmembrane-bound, ribonucleoprotein organelles called germ granules. Depletion of germ granules in Caenorhabditis elegans (i.e., P granules) leads to sterility and, in some germlines, expression of the neuronal transgene unc-119::gfp and the muscle myosin MYO-3. Thus, P granules are hypothesized to maintain germ cell totipotency by preventing somatic development, although the mechanism by which P granules carry out this function is unknown. In this study, we performed transcriptome and single molecule RNA-FISH analyses of dissected P granule-depleted gonads at different developmental stages. Our results demonstrate that P granules are necessary for adult germ cells to downregulate spermatogenesis RNAs and to prevent the accumulation of numerous soma-specific RNAs. P granule-depleted gonads that express the unc-119::gfp transgene also express many other genes involved in neuronal development and concomitantly lose expression of germ cell fate markers. Finally, we show that removal of either of two critical P-granule components, PGL-1 or GLH-1, is sufficient to cause germ cells to express UNC-119::GFP and MYO-3 and to display RNA accumulation defects similar to those observed after depletion of P granules. Our data identify P granules as critical modulators of the germline transcriptome and guardians of germ cell fate.

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Section: Discussionmentioning

confidence: 99%

Germ Granules Prevent Accumulation of Somatic Transcripts in the AdultCaenorhabditis elegansGermline

et al. 2017

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“…The extrachromosomal array was integrated using gamma-ray irradiation and outcrossed four times to WT N2. Strain QW1217 was made by combining zfIs124 with otIs355(Prab-3::NLS::tagRFP) (gift of O. Hobert) (21). Strain QW1473 (rgef::GFP and rab3::NLS-tagRFP) was made by combining strain evIs111(Prgef::GFP;dpy-20) with otIs355(Prab-3::NLS::tagRFP) (46).…”

Section: Methodsmentioning

confidence: 99%

“…Activity was reported by the ultrasensitive calcium indicator GCaMP6s expressed throughout the cytosol under the control of the pan-neuronal rgef-1 promoter (a gift from D. Pilgrim, University of Alberta, Edmonton, Alberta, Canada) (20). To facilitate segmentation into individual identifiable neurons, nuclei were tracked using calcium-insensitive, nuclear-bound red fluorescent protein (RFP), TagRFP, under the control of another pan-neuronal rab-3 promoter (a gift from O. Hobert, Columbia University, New York) (21). We developed an image analysis pipeline that converts the gross movements of the head into the time-varying position and posture of the crawling worm, and converts fluorescence measurements into near simultaneous activity patterns of all imaged neurons.…”

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confidence: 99%

Pan-neuronal imaging in roaming Caenorhabditis elegans

Venkatachalam

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

221

216

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We present an imaging system for pan-neuronal recording in crawling Caenorhabditis elegans. A spinning disk confocal microscope, modified for automated tracking of the C. elegans head ganglia, simultaneously records the activity and position of ∼80 neurons that coexpress cytoplasmic calcium indicator GCaMP6s and nuclear localized red fluorescent protein at 10 volumes per second. We developed a behavioral analysis algorithm that maps the movements of the head ganglia to the animal's posture and locomotion. Image registration and analysis software automatically assigns an index to each nucleus and calculates the corresponding calcium signal. Neurons with highly stereotyped positions can be associated with unique indexes and subsequently identified using an atlas of the worm nervous system. To test our system, we analyzed the brainwide activity patterns of moving worms subjected to thermosensory inputs. We demonstrate that our setup is able to uncover representations of sensory input and motor output of individual neurons from brainwide dynamics. Our imaging setup and analysis pipeline should facilitate mapping circuits for sensory to motor transformation in transparent behaving animals such as C. elegans and Drosophila larva.C. elegans | Drosophila | calcium imaging | thermotaxis U nderstanding how brain dynamics creates behaviors requires quantifying the flow and transformation of sensory information to motor output in behaving animals. Optical imaging using genetically encoded calcium or voltage fluorescent probes offers a minimally invasive method to record neural activity in intact animals. The nematode Caenorhabditis elegans is particularly ideal for optical neurophysiology owing to its small size, optical transparency, compact nervous system, and ease of genetic manipulation. Imaging systems for tracking the activity of small numbers of neurons have been effective in determining their role during nematode locomotion and navigational behaviors like chemotaxis, thermotaxis, and the escape response (1-6).Recordings from large numbers of interconnected neurons are required to understand how neuronal ensembles carry out the systematic transformations of sensory input into motor patterns that build behavioral decisions.Several methods for fast 3D imaging of neural activity in a fixed imaging volume have been developed for different model organisms (7)(8)(9)(10)(11)(12)(13)(14). High-speed light sheet microscopy, light field microscopy, multifocus microscopy, and two-photon structured illumination microscopy have proved effective for rapidly recording large numbers of neurons in immobilized, intact, transparent animals like larval zebrafish and nematodes (15)(16)(17)(18)(19). However, these methods are problematic when attempting to track many neurons within the bending and moving body of a behaving animal. Panneuronal recording in moving animals poses higher demands on spatial and temporal resolution. Furthermore, extracting neuronal signals from recordings in a behaving animal requires an effective analysis pipel...

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“…Furthermore, a fundamental characteristic of germ cells is their totipotency, allowing the zygote to differentiate into all somatic tissues after fertilization. Ectopic germline expression of master transcription factors necessary for speci fi c somatic differentiation programs, or the elimination of certain post-transcriptional regulators leads to a loss of germ cell fate identity and the formation of neuronal, muscle, or gut cells in the germ line (Ciosk et al 2006 ;Tursun et al 2011 ) . It is conceivable that general transcriptional activity is tightly controlled to avoid unwanted cell fate commitment.…”

Section: Introductionmentioning

confidence: 99%

Translational Control in the Caenorhabditis elegans Germ Line

Nousch

Eckmann

2012

Advances in Experimental Medicine and Biology

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Translational control is a prevalent form of gene expression regulation in the Caenorhabditis elegans germ line. Linking the amount of protein synthesis to mRNA quantity and translational accessibility in the cell cytoplasm provides unique advantages over DNA-based controls for developing germ cells. This mode of gene expression is especially exploited in germ cell fate decisions and during oogenesis, when the developing oocytes stockpile hundreds of different mRNAs required for early embryogenesis. Consequently, a dense web of RNA regulators, consisting of diverse RNA-binding proteins and RNA-modifying enzymes, control the translatability of entire mRNA expression programs. These RNA regulatory networks are tightly coupled to germ cell developmental progression and are themselves under translational control. The underlying molecular mechanisms and RNA codes embedded in the mRNA molecules are beginning to be understood. Hence, the C. elegans germ line offers fertile grounds for discovering post-transcriptional mRNA regulatory mechanisms and emerges as great model for a systems level understanding of translational control during development.

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Direct Conversion of C. elegans Germ Cells into Specific Neuron Types

Cited by 215 publications

References 29 publications

Germ Granules Prevent Accumulation of Somatic Transcripts in the AdultCaenorhabditis elegansGermline

Germ Granules Prevent Accumulation of Somatic Transcripts in the AdultCaenorhabditis elegansGermline

Pan-neuronal imaging in roaming Caenorhabditis elegans

Translational Control in the Caenorhabditis elegans Germ Line

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