The mammalian sperm must be highly motile for a long time to fertilize egg. It has been supposed that ATP required flagellar movement depends predominantly on mitochondrial respiration. We assessed the contribution of respiration mouse motility. Mouse maintained vigorous motility with high beat frequency in an appropriate solution including substrate such as glucose. active contained large amount ATP. When carbonyl cyanide m-chlorophenylhydrazone (CCCP) was applied suppress oxidative...

Highlights•CaV2.3 mediates spatiotemporal regulation of acrosome exocytosis (AE)•GM1 regulates Ca2+ influx, AE, and fertilization via CaV2.3's α1E α2δ1 subunits•Mechanism involves GM1's lipid sugar moieties α2δ1SummaryMembrane cell function is poorly understood. In early development, sterol efflux the ganglioside GM1 regulate sperm (AE) competence through unknown mechanisms. Here, we show that focal enrichment trigger influx necessary for AE CaV2.3, whose activity has been highly...

Development of assisted reproductive technologies (ART) in the dog has resisted progress for decades, due to their unique physiology. This lack is remarkable given critical role ART could play conserving endangered canid species or eradicating heritable disease through gene-editing technologies—an approach that would also advance as a biomedical model. Over 350 disorders/traits dogs are homologous with human conditions, almost twice number any other species. Here we report first live births...

Many studies have been conducted to elucidate the relationship between energy metabolic pathways (glycolysis and respiration) flagellar motility in mammalian sperm, but contribution of glycolysis sperm has not yet fully elucidated. In present study, we performed detailed analysis mouse for further understanding motility. Mouse maintained vigorous presence substrates either or respiration. By contrast, inhibition by alpha-chlorohydrine caused a significant decrease bend angle bending wave,...

Function of the mammalian translocator protein (TSPO; previously known as peripheral benzodiazepine receptor) remains unclear because its presumed role in steroidogenesis and mitochondrial permeability transition established using pharmacological methods has been refuted recent genetic studies. Protoporphyrin IX (PPIX) is considered a conserved endogenous ligand for TSPO. In bacteria, TSPO was identified to regulate tetrapyrrole metabolism chemical catalysis PPIX presence light, vertebrates,...

SUMMARY Salmonid fish sperm motility is known to be suppressed in millimolar concentrations of extracellular K+, and dilution K+ upon spawning triggers cAMP-dependent signaling for initiation. In a previous study, however, we demonstrated that suspending 10% glycerol solution subsequent into low-osmotic induced independently cAMP. the present further investigated glycerol-induced mechanism. We found treatment with solutions consisting organic or inorganic ions, as well glycerol, an...

Abstract For nanobiotechnology to achieve its potential, complex organic–inorganic systems must grow utilize the sequential functions of multiple biological components. Critical challenges exist: immobilizing enzymes can block substrate‐binding sites or prohibit conformational changes, substrate composition interfere with activity, and multistep reactions risk diffusion intermediates. As a result, most tethered reaction reported involves only 3 enzymes. Inspired by oriented immobilization...

Abstract Background Rearranged during transfection (RET) tyrosine kinase signaling has been previously implicated in endocrine resistant breast cancer, however the mechanism by which this cascade promotes resistance is currently not well described. We recently reported that glial cell-derived neurotrophic factor (GDNF)-RET appears to promote a positive feedback loop with transcription early growth response 1 (EGR1). Here we investigate behind and test hypothesis GDNF-RET forms regulatory...

Despite numerous applications, we lack fundamental understanding of how variables such as nanoparticle (NP) size influence the activity tethered enzymes. Previously, showed that biomimetic oriented immobilization yielded higher specific activities versus nonoriented adsorption or carboxyl-amine binding. Here, standardize NP attachment strategy (oriented via hexahistidine tags) and composition (Ni-NTA coated gold NPs), to test impact (⌀5, 10, 20, 50 nm) on multilayer formation, activity,...

The RET tyrosine kinase signaling pathway is involved in the development of endocrine resistant ER+ breast cancer. However, we know little about how cells activate and initiate an phenotype. Here show that both sensitive cancers have a functional pathway, but cancer lack ligands are necessary to drive resistance. Transcription one ligand, GDNF, sufficient confer resistance MCF-7 cell line. Endogenous GDNF produced by translated, secreted into media, activates nearby cells. In patients,...

Secretion of the acrosome, a single vesicle located rostrally in head mammalian sperm, through process known as "acrosome exocytosis" (AE), is essential for fertilization. However, mechanisms leading to and regulating this complex are controversial. In particular, poor understanding Ca2+ dynamics between sperm subcellular compartments regulation membrane fusion have led competing models AE. Here, we developed transgenic mouse expressing an Acrosome-targeted Sensor Exocytosis (AcroSensE)...

Abstract The murine epididymis has 10 distinct segments that provide the opportunity to identify compartmentalized cell physiological mechanisms underlying sperm maturation. However, despite essential role of in reproduction, remarkably little is known about segment-specific functions this organ. Here, we investigate dramatic segmental localization ganglioside GM1, a glycosphingolipid already play key roles capacitation and acrosome exocytosis. Frozen tissue sections epididymides from adult...

Tenascin and fibronectin are components of the extracellular matrices that oppose promote adhesion, respectively. Using immunohistochemical techniques, we studied distribution tenascin in mouse ovary, which dynamic reconstruction degeneration occur during folliculogenesis, atresia, ovulation, corpus luteum formation luteolysis. In growing follicles, was only detected theca externa layer, while interna layer basement membrane. During follicular granulosa cells, surrounded by membrane, began...

Maintaining activity of enzymes tethered to solid interfaces remains a major challenge in developing hybrid organic-inorganic devices. In nature, mammalian spermatozoa have overcome this design by having glycolytic with specialized targeting domains that enable them function while cytoskeletal element. As step toward designing ATP-generating system, we implemented biomimetic site-specific immobilization strategy tether two representing different functional enzyme families: triose...

Background Rapid diagnosis for time-sensitive illnesses such as stroke, cardiac arrest, and septic shock is essential successful treatment. Much attention has therefore focused on new strategies rapid objective diagnosis, Point-of-Care Tests (PoCT) blood biomarkers. Here we use a biomimicry-based approach to demonstrate diagnostic platform, based enzymes tethered nanoparticles (NPs). As proof of principle, oriented immobilization pyruvate kinase (PK) luciferase (Luc) silica NPs achieve...

Peptidylarginine deiminase enzymes (PADs) convert arginine residues to citrulline in a process called citrullination or deimination. Recently, two PADs, PAD2 and PAD4, have been linked hormone signaling vitro the goal of this study was test for links between PAD2/PAD4 vivo.Preliminary analysis Padi2 Padi4 single knockout (SKO) mice did not find any overt reproductive defects we predicted that likely due genetic compensation. To hypothesis, created Padi2/Padi4 double (DKO) mouse model tested...

Acrosome exocytosis (AE), in which the sperm's single exocytotic vesicle fuses with plasma membrane, is a complex, calcium-dependent process essential for fertilization. However, our understanding of how calcium signaling regulates AE still incomplete. In particular, interplay between intra-acrosomal dynamics and intermediate steps leading to not well-defined. Here, we describe method that provides spatial temporal insights into acrosomal their relationship membrane fusion subsequent...

Abstract Background Canine visceral hemangiosarcoma (HSA) is a highly aggressive cancer of endothelial origin that closely resembles angiosarcoma in humans, both clinically and histopathologically. Currently there an unmet need for new diagnostics therapies forms this disease. The goal study was to utilize Chromatin run-on sequencing (ChRO-seq) immunohistochemistry (IHC) identify gene protein expression signatures may be important drivers HSA progression. Results ChRO-seq performed on tissue...

Discovering regulatory interactions between genes that specify the behavioral properties of cells remains an important challenge. We used dynamics transcriptional changes resolved by PRO-seq to identify a network responsible for endocrine resistance in breast cancer. show GDNF leads switching active state bi-stable feedback loop GDNF, EGR1, and master transcription factor ERα. stimulates MAP kinase, activating factors SRF AP-1. initiates immediate response, EGR1 suppressing Newly translated...