Abstract:Spermatogonial stem cells (SSCs) provide the basis for lifelong male fertility through self-renewal and differentiation. Prepubertal male cancer patients may be rendered infertile by gonadotoxic chemotherapy and, unlike sexually mature men, cannot store sperm. Alternatively, testicular biopsies taken prior to treatment may be used to restore fertility in adulthood. Testicular SSC populations are limited, and in vitro culture systems are required to increase numbers of SSCs for treatment, demanding culture syst… Show more
“…SCs-derived sEVs were labeled by PKH26 and incubated with SSCLCs for 24 h. Red fluorescence staining was observed in SSCLCs while control group was not (Fig. 6 B), suggesting that sEVs derived from SCs could be taken in by SSCLCs, which had also been testified by other researchers previously [ 31 , 32 ]. Fig.…”
Section: Resultssupporting
confidence: 77%
“…Moreover, the sEVs released by SCs have emerged as vital components in intercellular communication between SC and SSC [ 29 , 30 ]. Researchers found that inhibition of SC derived-sEVs reduced spermatogonial proliferation and also alter the differentiation of SSCs in vitro [ 31 , 32 ]. The SC-SSC communication is a representative intercellular model that explains the role of sEVs in signal transduction.…”
Background
Small extracellular vesicles (EVs), exemplified by exosomes, mediate intercellular communication by transporting proteins, mRNAs, and miRNAs. Post-translational modifications are involved in controlling small EV secretion process. However, whether palmitoylation regulates small EV secretion, remains largely unexplored.
Methods
Vacuole Membrane Protein 1 (VMP1) was testified to be S-palmitoylated by Palmitoylation assays. VMP1 mutant plasmids were constructed to screen out the exact palmitoylation sites. Small EVs were isolated, identified and compared between wild-type VMP1 or mutant VMP1 transfected cells. Electron microscope and immunofluorescence were used to detect multivesicular body (MVB) number and morphology change when VMP1 was mutated. Immunoprecipitation and Mass spectrum were adopted to identify the protein that interacted with palmitoylated VMP1, while knock down experiment was used to explore the function of targeted protein ALIX. Taking human Sertoli cells (SCs) and human spermatogonial stem cell like cells (SSCLCs) as a model of intercellular communication, SSCLC maintenance was detected by flow cytometry and qPCR at 12 days of differentiation. In vivo, mouse model was established by intraperitoneal injection with palmitoylation inhibitor, 2-bromopalmitate (2BP) for 3 months.
Results
VMP1 was identified to be palmitoylated at cysteine 263,278 by ZDHHC3. Specifically, palmitoylation of VMP1 regulated its subcellular location and enhanced the amount of small EV secretion. Mutation of VMP1 palmitoylation sites interfered with the morphology and biogenesis of MVBs through suppressing intraluminal vesicle formation. Furthermore, inhibition of VMP1 palmitoylation impeded small EV secretion by affecting the interaction of VMP1 with ALIX, an accessory protein of the ESCRT machinery. Taking SCs and SSCLCs as a model of intercellular communication, we discovered VMP1 palmitoylation in SCs was vital to the growth status of SSCLCs in a co-culture system. Inhibition of VMP1 palmitoylation caused low self-maintenance, increased apoptosis, and decreased proliferation rate of SSCLCs. In vivo, intraperitoneal injection of 2BP inhibited VMP1 palmitoylation and exosomal marker expression in mouse testes, which were closely associated with the level of spermatogenic cell apoptosis and proliferation.
Conclusions
Our study revealed a novel mechanism for small EV secretion regulated by VMP1 palmitoylation in Sertoli cells, and demonstrated its pivotal role in intercellular communication and SSC niche.
“…SCs-derived sEVs were labeled by PKH26 and incubated with SSCLCs for 24 h. Red fluorescence staining was observed in SSCLCs while control group was not (Fig. 6 B), suggesting that sEVs derived from SCs could be taken in by SSCLCs, which had also been testified by other researchers previously [ 31 , 32 ]. Fig.…”
Section: Resultssupporting
confidence: 77%
“…Moreover, the sEVs released by SCs have emerged as vital components in intercellular communication between SC and SSC [ 29 , 30 ]. Researchers found that inhibition of SC derived-sEVs reduced spermatogonial proliferation and also alter the differentiation of SSCs in vitro [ 31 , 32 ]. The SC-SSC communication is a representative intercellular model that explains the role of sEVs in signal transduction.…”
Background
Small extracellular vesicles (EVs), exemplified by exosomes, mediate intercellular communication by transporting proteins, mRNAs, and miRNAs. Post-translational modifications are involved in controlling small EV secretion process. However, whether palmitoylation regulates small EV secretion, remains largely unexplored.
Methods
Vacuole Membrane Protein 1 (VMP1) was testified to be S-palmitoylated by Palmitoylation assays. VMP1 mutant plasmids were constructed to screen out the exact palmitoylation sites. Small EVs were isolated, identified and compared between wild-type VMP1 or mutant VMP1 transfected cells. Electron microscope and immunofluorescence were used to detect multivesicular body (MVB) number and morphology change when VMP1 was mutated. Immunoprecipitation and Mass spectrum were adopted to identify the protein that interacted with palmitoylated VMP1, while knock down experiment was used to explore the function of targeted protein ALIX. Taking human Sertoli cells (SCs) and human spermatogonial stem cell like cells (SSCLCs) as a model of intercellular communication, SSCLC maintenance was detected by flow cytometry and qPCR at 12 days of differentiation. In vivo, mouse model was established by intraperitoneal injection with palmitoylation inhibitor, 2-bromopalmitate (2BP) for 3 months.
Results
VMP1 was identified to be palmitoylated at cysteine 263,278 by ZDHHC3. Specifically, palmitoylation of VMP1 regulated its subcellular location and enhanced the amount of small EV secretion. Mutation of VMP1 palmitoylation sites interfered with the morphology and biogenesis of MVBs through suppressing intraluminal vesicle formation. Furthermore, inhibition of VMP1 palmitoylation impeded small EV secretion by affecting the interaction of VMP1 with ALIX, an accessory protein of the ESCRT machinery. Taking SCs and SSCLCs as a model of intercellular communication, we discovered VMP1 palmitoylation in SCs was vital to the growth status of SSCLCs in a co-culture system. Inhibition of VMP1 palmitoylation caused low self-maintenance, increased apoptosis, and decreased proliferation rate of SSCLCs. In vivo, intraperitoneal injection of 2BP inhibited VMP1 palmitoylation and exosomal marker expression in mouse testes, which were closely associated with the level of spermatogenic cell apoptosis and proliferation.
Conclusions
Our study revealed a novel mechanism for small EV secretion regulated by VMP1 palmitoylation in Sertoli cells, and demonstrated its pivotal role in intercellular communication and SSC niche.
“…The cargo of Sertoli cell‐derived exosomes has been a subject of extensive research, shedding light on their potential implications in spermatogenesis and beyond. The exchange of EVs, including exosomes, between SCs and other cell types within the testicular microenvironment is essential for supporting spermatogenesis and maintaining testicular homeostasis (Thiageswaran et al., 2022). The role of Sertoli cell‐derived exosomes in spermatogenesis is of significant interest in understanding the mechanisms involved in male fertility and reproductive capacity.…”
Section: Exosomes As Therapeutic Tools In Male Infertilitymentioning
BackgroundSpermatogenesis is a fundamental process crucial for male reproductive health and fertility. Exosomes, small membranous vesicles released by various cell types, have recently garnered attention for their role in intercellular communication.ObjectiveThis review aims to comprehensively explore the role of exosomes in regulating spermatogenesis, focusing on their involvement in testicular development and cell‐to‐cell communication.MethodsA systematic examination of literature was conducted to gather relevant studies elucidating the biogenesis, composition, and functions of exosomes in the context of spermatogenesis.ResultsExosomes play a pivotal role in orchestrating the complex signaling networks required for proper spermatogenesis. They facilitate the transfer of key regulatory molecules between different cell populations within the testes, including Sertoli cells, Leydig cells, and germ cells.ConclusionThe emerging understanding of exosome‐mediated communication sheds light on novel mechanisms underlying spermatogenesis regulation. Further research in this area holds promise for insights into male reproductive health and potential therapeutic interventions.
“…In an experiment conducted by Thiageswaran and co‐workers, they proved that the inhibition of Exo biogenesis in Sertoli cells can reduce the proliferation capacity of porcine spermatogonial stem cells in the co‐culture system. [ 52 ] Exos can be uptaken via sperms within the interstitial space and seminiferous tubules. [ 53 ] Like Sertoli cells, Leydig cells and local macrophages of testicular tissue can produce Exos in which selected non‐coding RNAs can be found in sperm transcriptome payload.…”
Section: Exos and The Male Reproductive Systemmentioning
Exosomes (Exos), belonging to extracellular vesicles, are cell‐derived nano‐sized vesicles with the potential to carry different kinds of biological molecules. Many studies have proved the impacts of exosomal cargo on several biological processes in female and male reproductive systems. It is also hypothesized that changes in exosomal cargo are integral to the promotion of certain pathological conditions, thus Exos can be used as valid biomarkers for the diagnosis of infertility and other abnormal conditions. Here, efforts are made to collect some recent data related to the physiological significance of Exos in the reproductive system, and their potential therapeutic effects. It is anticipated that the current review article will lay the groundwork for elucidating the source and mechanisms by which Exos control the reproductive system additionally supplying fresh methods and concepts for the detection and treatment of disorders associated with fertility for future studies.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
