Endocytic recycling in plants: pathways and regulation.

Endocytic recycling is an intracellular trafficking pathway that returns endocytosed molecules to the plasma membrane via the recycling endosome. This pathway plays a crucial role in remodelling plasma membrane composition and is thus essential for cellular homeostasis. In plants, endocytic recycling regulates the localization and abundance of receptors, transporters, and channels at the plasma membrane that are involved in many aspects of plant growth and development. Despite its importance, the recycling endosome and the underlying sorting mechanisms for cargo recycling in plants remain understudied in comparison to the endocytic recycling pathways in animals. In this review, we focus on the cumulative evidence suggesting the existence of endosomes decorated by regulators that contribute to recycling in plant cells. We summarize the chemical inhibitors used for analysing cargo recycling and discuss recent advances in our understanding of how endocytic recycling participates in various plant cellular and physiological events.

Circular RNAs in non-alcoholic fatty liver disease: Functions and clinical significance.

Nonalcoholic fatty liver disease (NAFLD), which affects approximately 25% of the global population, is an urgent health issue leading to various metabolic comorbidities. Circular RNAs (circRNAs), covalently closed RNA molecules, are characterized by ubiquity, diversity, stability, and conservatism. Indeed, they participate in various biological processes via distinct mechanisms that could modify the natural history of NAFLD. In this review, we briefly introduce the biogenesis, characteristics, and biological functions of circRNAs. Furthermore, we summarize circRNAs expression profiles in NAFLD by intersecting seven sequencing data sets and describe the cellular roles of circRNAs and their potential advantages as biomarkers of NAFLD. In addition, we emphatically discuss the exosomal non-coding RNA sorting mechanisms and possible functions in recipient cells. Finally, we extensively discuss the potential application of targeting disease-related circRNAs and competing endogenous RNA networks through gain-of-function and loss-of-function approaches in targeted therapy of NAFLD.

Extracellular Vesicle MicroRNAs in Heart Failure: Pathophysiological Mediators and Therapeutic Targets.

Extracellular vesicles (EVs) are emerging mediators of intracellular and inter-organ communications in cardiovascular diseases (CVDs), especially in the pathogenesis of heart failure through the transference of EV-containing bioactive substances. microRNAs (miRNAs) are contained in EV cargo and are involved in the progression of heart failure. Over the past several years, a growing body of evidence has suggested that the biogenesis of miRNAs and EVs is tightly regulated, and the sorting of miRNAs into EVs is highly selective and tightly controlled. Extracellular miRNAs, particularly circulating EV-miRNAs, have shown promising potential as prognostic and diagnostic biomarkers for heart failure and as therapeutic targets. In this review, we summarize the latest progress concerning the role of EV-miRNAs in HF and their application in a therapeutic strategy development for heart failure.

Roles of long noncoding RNAs and small extracellular vesicle-long noncoding RNAs in type 2 diabetes.

The prevalence of a high-energy diet and a sedentary lifestyle has increased the incidence of type 2 diabetes (T2D). T2D is a chronic disease characterized by high blood glucose levels and insulin resistance in peripheral tissues. The pathological mechanism of this disease is not fully clear. Accumulated evidence has shown that noncoding RNAs have an essential regulatory role in the progression of diabetes and its complications. The roles of small noncoding RNAs, such as miRNAs, in T2D, have been extensively investigated, while the function of long noncoding RNAs (lncRNAs) in T2D has been unstudied. It has been reported that lncRNAs in T2D play roles in the regulation of pancreatic function, peripheral glucose homeostasis and vascular inflammation. In addition, lncRNAs carried by small extracellular vesicles (sEV) were shown to mediate communication between organs and participate in diabetes progression. Some sEV lncRNAs derived from stem cells are being developed as potential therapeutic agents for diabetic complications. In this review, we summarize the current knowledge relating to lncRNA biogenesis, the mechanisms of lncRNA sorting into sEV and the regulatory roles of lncRNAs and sEV lncRNAs in diabetes. Knowledge of lncRNAs and sEV lncRNAs in diabetes will aid in the development of new therapeutic drugs for T2D in the future.

No size-dependent net particle retention in the hindgut of horses.

Sieve analyses of hindgut contents of horses as well as observations in horses where plastic markers had been applied to a caecal cannula suggested that there may be a discrimination by particle size in the passage or retention of digesta. Here, we performed a similar experiment with five caecum-cannulated horses (562 ± 31 kg) fed a constant amount (6.81 kg dry matter/day) of grass hay. Passage markers representing the liquid (Co-EDTA) as well as the particulate digesta phase (Yb-undefined; Cr mordanted fibre 1-2 mm; Ce-mordanted fibre 8 mm) were given as a pulse-dose into the cannula to measure their mean retention times (MRT). The MRTs were compared by repeated-measurements analysis of variance. The MRT in the hindgut was 22.2 ± 2.4 h for Co, 25.0 ± 3.4 h for Yb, 26.2 ± 1.6 h for Cr and 26.3 ± 1.5 h for Ce. Whereas differences between the particle marker MRTs were not significant (padj. > 0.05), significant differences were observed between the solute marker Co and each of the particle markers Cr and Ce (padj. < 0.009). The results confirm the well-known significant, albeit small, difference in MRT in horses between the fluid and the particle digesta phase, and corroborate another recent study that used a combination of whole, marked hay and individual marker analysis in different particle size fractions of the faeces, which also did not detect a selective retention of any particle size class.

Emerging role of exosomal long non-coding RNAs in lung cancer.

BACKGROUND: Lung cancer is one of the most common malignancies worldwide. Also, it is the leading cause of cancer morbidity and mortality in men. Despite advances in lung cancer diagnosis and treatment, novel approaches are strongly needed to promote early diagnosis and effective treatment of lung cancer. Presently, accumulating data reveal that long noncoding RNAs (lncRNAs) are differentially enriched in exosomes and mediate multiple biological processes in lung cancer, suggesting the potential application of exosomal lncRNAs as diagnostic biomarkers and therapeutic targets. CONCLUSION: In this review, we described the emerging roles of lncRNAs specifically sorted into exosomes in lung cancer. We discussed the current knowledge of the exosomal lncRNA sorting mechanism and highlighted opportunities for exosome-derived lncRNAs as biomarkers in clinical practice. In particular, we systematically summarized the biological functions of exosomal lncRNAs in lung cancer.

Corrigendum: Insights Into Exosomal Non-Coding RNAs Sorting Mechanism and Clinical Application.

[This corrects the article DOI: 10.3389/fonc.2021.664904.].

Heterogeneous Nuclear Ribonucleoprotein A1 Loads Batched Tumor-Promoting MicroRNAs Into Small Extracellular Vesicles With the Assist of Caveolin-1 in A549 Cells.

MicroRNAs in small extracellular vesicle (sEV-miRNAs) have been widely investigated as crucial regulated molecules secreted by tumor cells to communicate with surroundings. It is of great significance to explore the loading mechanism of sEV-miRNAs by tumor cells. Here, we comprehensively illustrated a reasoned loading pathway of batched tumor-promoting sEV-miRNAs in non-small cell lung cancer (NSCLC) cell line A549 with the application of a multi-omics method. The protein heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) was strictly selected as a powerful sEV-miRNA loading protein from miRNA-binding proteome and further verified through small RNA sequencing after hnRNPA1 silence. In terms of the mechanism, SUMOylated hnRNPA1 in sEVs was verified to control sEV-miRNA loading. Subsequently, as a scaffolding component of caveolae, caveolin-1 (CAV1) was detailedly demonstrated to assist the loading of SUMOylated hnRNPA1 and its binding miRNAs into sEVs. Inhibition of CAV1 significantly prevented SUMOylated hnRNPA1 from encapsulating into sEVs, resulting in less enrichment of sEV-miRNAs it loaded. Finally, we confirmed that hnRNPA1-loaded sEV-miRNAs could facilitate tumor proliferation and migration based on database analysis and cytological experiments. Our findings reveal a loading mechanism of batched tumor-promoting sEV-miRNAs, which may contribute to the selection of therapeutic targets for lung cancer.

Insights Into Exosomal Non-Coding RNAs Sorting Mechanism and Clinical Application.

Exosomes are natural nanoscale bilayer phospholipid vesicles that can be secreted by almost all types of cells and are detected in almost all types of body fluids. Exosomes are effective mediators of cell-cell signaling communication because of their ability to carry and transfer a variety of bioactive molecules, including non-coding RNAs. Non-coding RNAs have also been found to exert strong effects on a variety of biological processes, including tumorigenesis. Many researchers have established that exosomes encapsulate bioactive non-coding RNAs that alter the biological phenotype of specific target cells in an autocrine or a paracrine manner. However, the mechanism by which the producer cells package non-coding RNAs into exosomes is not well understood. This review focuses on the current research on exosomal non-coding RNAs, including the biogenesis of exosomes, the possible mechanism of sorting non-coding RNAs, their biological functions, and their potential for clinical application in the future.

Body weight and sex effects on digesta mean retention time in growing Saanen goats.

Despite the important role of digesta mean retention time (MRT) on digestive efficiency of ruminants, it is poorly investigated in total gastrointestinal tract (GIT) of growing ruminants, especially in goats. The objective of this study was to evaluate the effect of body weight (BW) and sex on GIT MRT of particles and solutes in growing Saanen goats. A dataset from two studies, comprising 103 individual records of castrated males (n = 36), females (n = 34), and intact males (n = 33) Saanen goats slaughtered at 15, 22, 30, 37, and 45 kg BW, was used. Goats were fed basically with total mixed ration composed by dehydrated corn plant (Zea mays) milled to pass a 10-mm screen, cracked corn grain, and soybean (Glycine max) meal. Variables evaluated were BW, feed intake, feed intake level, composition of ingested diet, wet weight of GIT tissues, wet digesta pool size, digesta composition (dry matter and neutral detergent fiber [NDF]), indigestible NDF:NDF ratio of ingested diet and GIT digesta, MRT of particles (MRTiNDF) and solutes (MRTCr), and reticulorumen selectivity factors (large particles/solutes). Reticulorumen, omasum, abomasum, small intestine, cecum, and colon-rectum segments were evaluated. The dataset was analyzed as mixed models considering sex, BW, and sex × BW interaction as fixed effects, and study and residual error as random effects. Sex did not affect MRTiNDF in any GIT segments. Females and intact males presented similar reticulorumen MRTCr (5.6 h; P = 0.92) and they presented lower reticulorumen MRTCr than castrated males (7.0; P ≤ 0.04). Total GIT MRTCr was similar between castrated males and females (15.7 h; P = 0.11) and between females and intact males (14.2 h; P = 0.76). Body weight (BW) did not affect MRTiNDF in reticulorumen and colon-rectum and total GIT MRTCr (P ≥ 0.11). Reticulorumen and omasum MRTCr increased as BW increased (P < 0.01), and abomasum MRTCr decreased as BW increased (P = 0.02). Feed intake, and wet tissues and wet pool size of all GIT segments increased as BW increased, except abomasum wet pool size (P ≤ 0.01). The mechanism related to sex effect on MRT has to be elucidated. Reticulorumen MRTiNDF and total GIT MRTCr were modulated by intake and capacity of reticulorumen and GIT, respectively. On the other hand, reticulorumen MRTCr seemed to be regulated by reticulo-omasal orifice opening and saliva secretion.

Role and application of exosomes-mediated miRNAs in the treatment of glioma / 中国组织工程研究

BACKGROUND: Exosomes have become more and more popular in the field of tumor research in recent years. They can carry a large number of bioactive molecules directly to the receptor cells and participate in the. information exchange between cells. As the highest content and most kinds of non-coding RNA in exosomes, miRNAs play an important role in the proliferation, invasion, drug resistance and immunity of glioma. In addition, miRNAs have potential application value in the auxiliary diagnosis, treatment orientation and prognosis evaluation of glioma. OBJECTIVE: To describe the mechanism of exosome miRNAs sorting, the regulatory role in gliomas and its clinical application, to provide literature and theoretical basis for further exploring the relationship between exosome miRNAs and gliomas, and to exert positive significance in the diagnosis and treatment of gliomas. METHODS: The first author searched CNKI, WanFang database, PubMed database and EI database. The key words were “glioma, exosomes, miRNAs, sorting mechanism, biomarkers” in Chinese and English. Totally 118 literatures were retrieved. According to the inclusion and exclusion criteria, and literature supplement, totally 52 literatures were selected for summary and induction, mainly regarding the sorting mechanism of exosome miRNAs, the regulatory role in glioma and clinical application. RESULTS AND CONCLUSION: At present, the effective treatment of gliomas cannot meet the clinical needs even though a variety of high-intensity combined treatment schemes are adopted in this study. In this case, it is an important supplement for the clinical treatment of gliomas to propose the application of exosomes-mediated miRNAs in gene therapy of gliomas. Exosomes-mediated miRNAs cannot only play a regulatory role in the occurrence and development of glioma, invasion and metastasis, radiochemotherapy resistance, and immune regulation, but also play a potential role as a biomarker in the grading diagnosis, prognosis evaluation, treatment and other aspects of glioma. This will provide a solid theoretical basis for us to further explore the relationship between exosome miRNAs and glioma, and also has a positive clinical significance in the innovative diagnosis and treatment of glioma.

Physical characteristics of gastrointestinal content of llama (Lama glama).

Changes in digesta dry matter (DM) and mean digesta particle size (MPS) along the gastrointestinal tract are well known in ruminants, but not in camelids. We collected digesta from the dorsal (d) and ventral (v) first forestomach compartment (C1), the second forestomach compartment (C2), three proximal segments and the subsequent glandular part of the third compartment (C3A-D), the caecum and the faeces twelve llamas (Lama glama). DM analysis indicates the presence of digesta stratification in the C1, the presence of fluid in the C2 to facilitate the sorting function of this compartment, the fluid-absorbing function of the proximal parts of the C3, the secretion of enzymes and digestive acids in the C3D, and the water-resorbing function of the lower intestinal tract. These findings illustrate the functional resemblance between the gastrointestinal tract of camelids and cattle-like ruminants (C1 equivalent to the rumen with stratified contents, C2 to the reticulum, C3A/B/C to the omasum and C3D to the abomasum). MPS analysis revealed a progressive reduction in MPS from the C1 to the distal C3. This gradual transition is different from the clear-cut threshold in ruminants between the reticulum and the omasum and had so far only been described in dromedaries (Camelus dromedarius). These findings indicate that regardless of the convergent property of rumination and resemblance of general mechanisms involved in contents stratification and particle sorting, differences between ruminants and camelids exist that could be interpreted as a more efficient functionality of the ruminant forestomach.

Sorting of a HaloTag protein that has only a signal peptide sequence into exocrine secretory granules without protein aggregation.

The mechanism involved in the sorting and accumulation of secretory cargo proteins, such as amylase, into secretory granules of exocrine cells remains to be solved. To clarify that sorting mechanism, we expressed a reporter protein HaloTag fused with partial sequences of salivary amylase protein in primary cultured parotid acinar cells. We found that a HaloTag protein fused with only the signal peptide sequence (Met(1)-Ala(25)) of amylase, termed SS25H, colocalized well with endogenous amylase, which was confirmed by immunofluorescence microscopy. Percoll-density gradient centrifugation of secretory granule fractions shows that the distributions of amylase and SS25H were similar. These results suggest that SS25H is transported to secretory granules and is not discriminated from endogenous amylase by the machinery that functions to remove proteins other than granule cargo from immature granules. Another reporter protein, DsRed2, that has the same signal peptide sequence also colocalized with amylase, suggesting that the sorting to secretory granules is not dependent on a characteristic of the HaloTag protein. Whereas Blue Native PAGE demonstrates that endogenous amylase forms a high-molecular-weight complex, SS25H does not participate in the complex and does not form self-aggregates. Nevertheless, SS25H was released from cells by the addition of a ß-adrenergic agonist, isoproterenol, which also induces amylase secretion. These results indicate that addition of the signal peptide sequence, which is necessary for the translocation in the endoplasmic reticulum, is sufficient for the transportation and storage of cargo proteins in secretory granules of exocrine cells.