Isolation, profiling, and tracking of extracellular vesicle cargo in Caenorhabditis elegans

Nikonorova, Inna A.; Wang, Juan; Cope, Alexander L; Tilton, Peter; Power, Kaiden M; Walsh, Jonathon; Akella, Jyothi S.; Ar, Krauchunas; Shah, Premal; Barr, Maureen M.

doi:10.1016/j.cub.2022.03.005

Cited by 30 publications

(36 citation statements)

References 88 publications

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“…In hermaphrodites, the parsimonious hypothesis is that NSPF proteins passively diffuse from the spermathecae, where activated sperm reside, to the uterus. The location of NSPF proteins within the uterus is similar to other proteins that males transfer, which originate in newly identified extracellular vesicles (Nikonorova et al 2022). While the NSPFs do not appear in these vesicles, it is possible that this alternative vesicle type also contributes to seminal fluid proteins, supporting our finding that nematodes employ a distinct mechanism for secreting seminal fluid proteins than either mammals or Drosophila (McGraw et al 2015).…”

Section: Discussionmentioning

confidence: 78%

A novel sperm-derived seminal fluid protein inCaenorhabditisnematodes

Kasimatis

Rehaluk

Rowe

et al. 2022

Preprint

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Post-insemination molecular interactions involving sperm are an ideal system for linking genotype, phenotype, and fitness. Nematode sperm contain subcellular vesicles known as membranous organelles (MOs) that fuse with the cell membrane upon sperm activation to release their soluble contents into the extracellular space. The second most abundant protein group in the MOs is the conserved Nematode-Specific Peptide family, group F (NSPF) gene family. We hypothesize that these proteins contribute to seminal fluid upon MO fusion and are part of post-insemination reproductive tract dynamics. We used experimental evolution to compete the wildtype allele against a deletion allele in 10 replicate populations. We calculated a significant mean selective disadvantage of 0.1% for the deletion allele, which indicated that the NSPF genes are an evolutionarily important component of male reproductive success. This conclusion was reinforced by qualitative trends from lower powered single-generation male fertility assays. We then characterized the functional location of NSPF proteins during fertilization using whole- worm immunostaining of a His-tagged nspf-1 transgene. Throughout male development, we found that NSPF presence and abundance was correlated with reproductive maturity in males, localizing to the seminal vesicle during L4 and to the spicules throughout adulthood. We confirmed that NSPF proteins are transferred to females during mating and hypothesize that proteins adhere to the spicules during transfer. NSPF proteins localize to the vulva and uterus lumen when transferred to mated females and in unmated adult hermaphrodites. These results suggest that the uterine localization of the NSPF proteins is likely a functional property of both male-derived sperm and self-sperm and not simply incidental to the point of transfer during mating. Our study demonstrates that nematodes use a novel mechanism for generating seminal fluid proteins and shows that the highly abundant NSPF proteins likely have a female-beneficial fitness effect.

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Section: Discussionmentioning

confidence: 78%

A novel sperm-derived seminal fluid protein inCaenorhabditisnematodes

Kasimatis

Rehaluk

Rowe

et al. 2022

Preprint

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“…These results indicate that polycystin EV biogenesis, targeting, and signaling may be co-regulated. By contrast, other ciliary EV cargoes do not colocalize with PKD-2 on the same ciliary EV even though these cargoes are co-expressed with lov-1 and pkd-2 in male-specific sensory neurons [25, 43]. Our EV proteomic analysis identified 2,888 cargo candidates and revealed a heterogeneous EV population.…”

Section: Discussionmentioning

confidence: 81%

Tracking N- and C-termini of C. elegans polycystin-1 reveals their distinct targeting requirements and functions in cilia and extracellular vesicles

Walsh

Wang

DeHart

et al. 2022

Preprint

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The cilium acts as an antenna receiving and sending signals, the latter via extracellular vesicles (EVs). In C. elegans and mammals, the Autosomal Dominant Polycystic Kidney Disease (ADPKD) gene products polycystin-1 (PC1) and polycystin-2 (PC2) localize to both cilia and EVs, act in the same genetic pathway, and function in a sensory capacity, suggesting ancient conservation. However, the functions of the polycystins on cilia and EVs remain enigmatic. We used our C. elegans model and endogenously fluorescent-tagged LOV-1/polycystin-1 to study LOV-1 processing, trafficking, transport, EV biogenesis, and function in living animals. Super resolution, real time imaging reveals that LOV-1 is processed into N-terminal and C-terminal forms via a conserved GPCR proteolytic site (GPS). The LOV-1 NTM is secreted into the extracellular matrix and not localized to ciliary tip EVs. In contrast, LOV-1 CTM and PKD-2 are co-trafficked, co-transported, and co-localized in cilia and on environmentally-released ciliary EVs. LOV-1 CTM requires PKD-2 for ciliary EV localization, while PKD-2 localizes to ciliary EVs independent of LOV-1. We find that LOV-1 but not PKD-2 is required for chemosensation of an ascaroside mating pheromone. These findings indicate that the polycystins LOV-1 and PKD-2 function together and independently and provide insight to how cargo selected and packaged in ciliary EVs.

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“…Nikonorova et al demonstrated the biological function of exosome-loaded ENPP1 in mediating intercellular communication, involving various physiological and pathological states [16]. In recent years, ENPP1 has been found to play an important role in immune responses to various stimuli [17].…”

Section: Introductionmentioning

confidence: 99%

“…Ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), also known as plasma cell glycoprotein 1 (PC-1), is a type II transmembrane glycoprotein with nucleotide pyrophosphatase and phosphodiesterase activities [ 15 ]. Nikonorova et al demonstrated the biological function of exosome-loaded ENPP1 in mediating intercellular communication, involving various physiological and pathological states [ 16 ]. In recent years, ENPP1 has been found to play an important role in immune responses to various stimuli [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of Downregulated Exosome-Associated Gene ENPP1 as a Novel Lipid Metabolism and Immune-Associated Biomarker for Hepatocellular Carcinoma

Peng

et al. 2022

Journal of Oncology

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Exosome plays an important role in the occurrence and development of tumors, such as hepatocellular carcinoma (LIHC). However, the functions and mechanisms of exosome-associated molecules in LIHC are still underexplored. Here, we investigated the role of the exosome-related gene ENPP1 in LIHC. Comprehensive bioinformatics from multiple databases revealed that ENPP1 was significantly downregulated in LIHC tissues. The patients with downregulated ENPP1 displayed a poor prognosis. Immunohistochemistry (IHC) was used to further confirm the downregulated ENPP1 in LIHC tissues. In addition, the coexpression network of ENPP1 was also explored to understand its roles in the underlying signaling pathways, including fatty acid degradation and the PPAR signaling pathway. Simultaneously, GSEA analysis indicated the potential roles of ENPP1 in the lipid metabolism-associated signaling pathways in the pathogenesis of LIHC, including fatty acid metabolism, fatty acid synthesis, and so on. Finally, immunological analysis indicated that ENPP1 might also be involved in multiple immune-related features, including immunoinhibitors, immunostimulators, and chemokines. Taken together, these findings could enhance our understanding of ENPP1 in LIHC pathogenesis and immune response and provide a new target for ENPP1-related immunotherapy in clinical treatment.

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Isolation, profiling, and tracking of extracellular vesicle cargo in Caenorhabditis elegans

Cited by 30 publications

References 88 publications

A novel sperm-derived seminal fluid protein inCaenorhabditisnematodes

A novel sperm-derived seminal fluid protein inCaenorhabditisnematodes

Tracking N- and C-termini of C. elegans polycystin-1 reveals their distinct targeting requirements and functions in cilia and extracellular vesicles

Identification of Downregulated Exosome-Associated Gene ENPP1 as a Novel Lipid Metabolism and Immune-Associated Biomarker for Hepatocellular Carcinoma

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