TY - JOUR
T1 - Cell-Specific Transcriptional Profiling of Ciliated Sensory Neurons Reveals Regulators of Behavior and Extracellular Vesicle Biogenesis
AU - Wang, Juan
AU - Kaletsky, Rachel
AU - Silva, Malan
AU - Williams, April
AU - Haas, Leonard A.
AU - Androwski, Rebecca J.
AU - Landis, Jessica N.
AU - Patrick, Cory
AU - Rashid, Alina
AU - Santiago-Martinez, Dianaliz
AU - Gravato-Nobre, Maria
AU - Hodgkin, Jonathan
AU - Hall, David H.
AU - Murphy, Coleen T.
AU - Barr, Maureen M.
N1 - Funding Information:
We thank WormBase; the National Bioresource Project for strains; Christina DeCoste for assistance with FACS; Ken Nguyen, Leslie Gunther, and Geoff Perumal for help with HPF-FS, embedding, and serial section protocols performed at Einstein; William Rice and Ed Eng at the New York Structural Biology Center (NYSBC) for help with electron tomography; Lillian Hunter for assistance with strain construction; members of the M.M.B. and C.T.M. labs and Rutgers C. elegans community for discussion and insight, more than we ever learned in school; Dr. Emily Troemel for critical reading of the manuscript; and Rutgers Genetics Department for sabbatical time and critical bridge funding. Use of the NYSBC facilities was supported by the Albert Einstein College of Medicine. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). This work was funded by NIH DK059418 and DK074746 (to M.M.B.), NIH OD 010943 (to D.H.H.), and the Medical Research Council (to M.G.-N. and J.H.).
Publisher Copyright:
© 2015 Elsevier Ltd. All rights reserved.
PY - 2015/12/21
Y1 - 2015/12/21
N2 - Cilia and extracellular vesicles (EVs) are signaling organelles [1]. Cilia act as cellular sensory antennae, with defects resulting in human ciliopathies. Cilia both release and bind to EVs [1]. EVs are sub-micron-sized particles released by cells and function in both short- and long-range intercellular communication. In C. Elegans and mammals, the autosomal dominant polycystic kidney disease (ADPKD) gene products polycystin-1 and polycystin-2 localize to both cilia and EVs, act in the same genetic pathway, and function in a sensory capacity, suggesting ancient conservation [2]. A fundamental understanding of EV biology and the relationship between the polycystins, cilia, and EVs is lacking. To define properties of a ciliated EV-releasing cell, we performed RNA-seq on 27 GFP-labeled EV-releasing neurons (EVNs) isolated from adult C. Elegans. We identified 335 significantly overrepresented genes, of which 61 were validated by GFP reporters. The EVN transcriptional profile uncovered new pathways controlling EV biogenesis and polycystin signaling and also identified EV cargo, which included an antimicrobial peptide and ASIC channel. Tumor-necrosis-associated factor (TRAF) homologs trf-1 and trf-2 and the p38 mitogen-activated protein kinase (MAPK) pmk-1 acted in polycystin-signaling pathways controlling male mating behaviors. pmk-1 was also required for EV biogenesis, independent of the innate immunity MAPK signaling cascade. This first high-resolution transcriptome profile of a subtype of ciliated sensory neurons isolated from adult animals reveals the functional components of an EVN.
AB - Cilia and extracellular vesicles (EVs) are signaling organelles [1]. Cilia act as cellular sensory antennae, with defects resulting in human ciliopathies. Cilia both release and bind to EVs [1]. EVs are sub-micron-sized particles released by cells and function in both short- and long-range intercellular communication. In C. Elegans and mammals, the autosomal dominant polycystic kidney disease (ADPKD) gene products polycystin-1 and polycystin-2 localize to both cilia and EVs, act in the same genetic pathway, and function in a sensory capacity, suggesting ancient conservation [2]. A fundamental understanding of EV biology and the relationship between the polycystins, cilia, and EVs is lacking. To define properties of a ciliated EV-releasing cell, we performed RNA-seq on 27 GFP-labeled EV-releasing neurons (EVNs) isolated from adult C. Elegans. We identified 335 significantly overrepresented genes, of which 61 were validated by GFP reporters. The EVN transcriptional profile uncovered new pathways controlling EV biogenesis and polycystin signaling and also identified EV cargo, which included an antimicrobial peptide and ASIC channel. Tumor-necrosis-associated factor (TRAF) homologs trf-1 and trf-2 and the p38 mitogen-activated protein kinase (MAPK) pmk-1 acted in polycystin-signaling pathways controlling male mating behaviors. pmk-1 was also required for EV biogenesis, independent of the innate immunity MAPK signaling cascade. This first high-resolution transcriptome profile of a subtype of ciliated sensory neurons isolated from adult animals reveals the functional components of an EVN.
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U2 - 10.1016/j.cub.2015.10.057
DO - 10.1016/j.cub.2015.10.057
M3 - Article
C2 - 26687621
AN - SCOPUS:84955652335
SN - 0960-9822
VL - 25
SP - 3232
EP - 3238
JO - Current Biology
JF - Current Biology
IS - 24
ER -