Final Stages Of Maturation Research Articles

Gonadal recrudescence and annual reproductive hormone pattern of captive female Spotted Scats (Scatophagus argus)

The Spotted Scat (Scatophagus argus), an important euryhaline fish inhabiting mangrove and coastal regions of Indo-Pacific waters, is both an ornamental and food fish in India. Detailed insight into maturation of Spotted Scat when maintained in aquaculture systems, therefore, needs to be elucidated. Lack of information on annual maturation dynamics of female scat collected from their natural habitat and reared in earthen ponds is the basis of this study. Oocytes were classified into five developmental stages: pre-vitellogenic, vitellogenic, late-vitellogenic, ripe, and follicular atresia. Ovarian maturity stages were subsequently categorized as immature (Stage 1), vitellogenesis (Stage 2), maturing (Stage 3), mature (Stage 4), and spent (Stage 5). In oocytes in primary, secondary and tertiary yolk stages, there are greater concentrations of E2 in vitellogenic females between March and June. Significant increases of E2, T, and 17-OHP paralleled the increase of diameter of late–vitellogenic oocytes in maturing females during July. The completion of vitellogenesis and initiation of germinal vesicle migration in the cytoplasm were evident in mature females (Stage 4) with a decreasing trend of sex steroids in and subsequent to the month of August. There were 50 % of oocytes in the final oocyte maturation stage (FOM) (490–620 μm) until completion of Stage 4 in September. The results of this study indicate there is complete ovarian maturation in female scats captured in their natural habitat and maintained in an earthen pond, which may be important information for hatchery management for induction of spawning of Spotted Scat in aquaculture systems.

Features of the reproductive system development in the installations of closed water supply system

The paper considers the functional changes in the reproductive function of sturgeon species of different ages and their hybrids. The dynamics of the development of gonads of females and males during the reproductive cycle is studied. According to the results of studies, it is shown that when the hydrological and hydrochemical regimes of cultivation are stabilized, spawning intervals are reduced, as well as the time of maturity. In this case, the physiological and biochemical status of females at the final stages of maturity is normal, which is confirmed by research results. The development of oocytes in females raised under regulated conditions takes place without any deviations.

Reproductive physiology of bonefishes (Albula spp.) across the Northwest Bahamas.

Bonefishes (Albula spp.) are classified within the superorder Elopomorpha, which is comprised of over 1000 species that share a unique leptocephalus larval stage. Bonefishes have a circum-tropical distribution, inhabiting inshore shallow water flats and gathering in presumptive nearshore pre-spawn aggregations (PSA) during spawning months. These fishes support economically important recreational fisheries and subsistence fisheries throughout their ranges, yet little is known regarding their reproductive biology. Analysis of oocyte development and nutrient composition, and sex and gonadotrophic hormone levels, was conducted on females sampled in Grand Bahama, Central Andros, and South Andros, The Bahamas, to assess their reproductive state. Fish collected from the flats habitats along all three islands exhibited four major reproductive phases (immature, developing, spawning capable, and regressing). In contrast, all females captured at presumptive PSA sites had eggs in the final stage of oocyte maturation, significantly higher levels of all reproductive hormones (17β-estradiol, testosterone, and LH), larger vitellogenic oocytes, and oocytes exhibiting germinal vesicle migration and germinal vesicle breakdown. In addition, monthly variability in hormone levels and spawning readiness between Grand Bahama and Andros PSAs suggest that peak spawning times may differ among regions. Fatty acid and free amino acid composition and profiles, with high proportions of docosahexaenoic acid, histidine, and taurine, suggest that these nutrients are not only relevant as energy reserves, but also help achieve buoyancy and osmoregulation of oocytes. This study expands upon our understanding of fish reproductive and developmental physiology, and indicates potential factors influencing the survival and recruitment of bonefishes.

Out-of-season artificial reproduction techniques of cultured female tongue sole (Cynoglossus semilaevis): Broodstock management, administration methods of hormone therapy and artificial fertilization

The present study aimed at the development of an effective and efficient artificial reproduction technique for cultured female tongue sole (Cynoglossus semilaevis) in out-of-season. After about four-month enriching cultivation of broodstocks, a series of experiments were conducted to develop a protocol for the hormonal induction of ovulation and artificial fertilization. The results showed that single injection of combination of luteinizing hormone releasing hormone analogue (LHRH-A3), human chorionic gonadotropin (HCG) and domperidone (DOM) is effective to induce ovulation at different final maturation stages (i.e., stage IV and V). The optimal single injection dosages of combined hormones were (0.15 μg LHRH-A3 + 4 IU HCG + 2 mg DOM) / kg BW and (0.05 μg LHRH-A3 + 2 IU HCG + 1 mg DOM) / kg BW for females at stage IV and V respectively. At the optimized dose of hormones, the fertilization and hatching rates were 44.6% and 80.4%, 30.4% and 63.4% for stage IV and V respectively. The lapsed time to ovulation was very predictable and ranged from 35.5 to 39.5 h post injection. Ovulated eggs in ovarian cavity should be stripped within 15 min in order to obtain the highest fertilization and hatching rates, otherwise the values decreased significantly thereafter. These researches are expected to provide improved implementation of artificial reproduction techniques for the production of fertilized eggs in the commercial aquaculture of tongue sole.

194 Extracellular vesicles derived from ampullary oviductal fluid improve developmental competence of bovine oocytes

Invivo, oocytes mature in the presence of follicular fluid (FF) and ampullary oviductal fluid (AOF), starting from premature to final maturation stages (3-6h postovulation). Extracellular vesicles (EVs) have been identified as important mediators of gamete/embryo-maternal interactions. They carry regulatory molecules, such as microRNAs and mRNAs. So far, the functionality of EVs derived from FF and AOF has not yet been determined. The aim of this study was to evaluate the effect of EVs derived from FF and AOF during invitro oocyte maturation (IVM) on the development and quality of resulting bovine embryos. Follicular fluid of a preovulatory follicle was collected by ovum pickup, and AOF was collected from the oviducts of slaughtered cows in early luteal phase. Then, EVs from FF and AOF were isolated by size exclusion chromatography and confirmed by western blot. The concentration of EVs was determined by NanoDrop and Nanoparticle tracking analysis. Integrity and size of EVs were assessed by electron microscopy. Bovine oocytes (n=1331) were allocated at random to five groups. In the control group (C), oocytes (n=347) were cultured for 22.5h in maturation medium (TCM-199 and epidermal growth factor 20ngmL−1) (MM). In the negative control group (NC), oocytes (n=331) were cultured for 18h in MM, and then transferred into fresh MM for 4.5h. In the FF group, oocytes (n=162) were cultured in MM supplemented with FF EVs (12.5µgmL−1) for 18h, and then transferred to MM for 4.5h. In the AOF group, oocytes (n=328) were cultured for 18h in EV-free MM, and then transferred to MM supplemented with AOF EVs (1.7µgmL−1) for 4.5h. In the FF+AOF group, oocytes (n=163) were cultured in MM supplemented with FF EVs for 18h, and then transferred into MM supplemented with AOF EVs for 4.5h. At 22.5h post-incubation, mature oocytes were fertilized invitro. At 21h post-insemination, presumptive zygotes were denuded and cultured in synthetic oviductal fluid with insulin-transferrin-selenium for 8 days. Cleavage rate and blastocyst rate were recorded at 48h and 8 days post-insemination, respectively. Blastocyst quality was assessed by differential apoptotic staining. The data were analysed using one-way ANOVA. Cleavage rate was not affected by the treatment, whereas blastocyst yield in the AOF group (48.75%) was significantly higher than that in the C (42.91%) and NC (37.72%) groups. In addition, embryos produced in the AOF group showed the highest number of trophectoderm cells and inner cell mass cells, and lowest number of apoptotic cells (P<0.05). Trophectoderm and inner cell mass cells were higher and apoptotic cells were lower in FF and FF+AOF compared with the C and NC groups. In conclusion, adding AOF EVs to oocyte MM in the last 4.5h of IVM improves quantity and quality of blastocysts. In addition, although FF EVs showed no effect on blastocyst yield, they improved blastocyst quality by increasing cell number.

Ontogeny of the specificity of gonadotropin receptors and gene expression in carp.

The pituitary gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are the principle endocrine drivers of reproductive processes in the gonads of jawed vertebrates. Canonically, FSH recruits and maintains selected ovarian follicles for maturation and LH induces the stages of germinal vesicle breakdown and ovulation. In mammals, LH and FSH specifically activate cognate G-protein-coupled receptors that affect the proteins involved in steroidogenesis, protein hormone synthesis, and gametogenesis. This dual-gonadotropin model also exists in some fish species, but not in all. In fact, due to their diverse number of species, extended number of ecological niches, and remarkably flexible reproductive strategies, fish are appropriate as models to understand the co-evolution of gonadotropins and their receptors. In this study, we cloned and characterized the expression profile over the final stages of ovarian maturation of carp (Cyprinus carpio) LHCGR and FSHR. Expression of both gonadotropin receptors increased in the later stage of early vitellogenesis, suggesting that both LH and FSH play a role in the development of mature follicles. We additionally tested the activation of cLHCGR and cFSHR using homologous and heterologous recombinant gonadotropins in order to gain insight into an evolutionary model of permissive gonadotropin receptor function. These data suggest that carp (Cyprinus carpio) gonad development and maturation depends on a specific gonadotropin profile that does not reflect the temporally distinct dual-gonadotropin model observed in salmonids or mammals, and that permissive gonadotropin receptor activation is a specific feature of Ostariophysi, not all teleosts.

Quantitative metabolomics approach reveals dynamics of primary metabolites in ‘Kinnow’ mandarin (C. nobilis × C. deliciosa) during advanced stages of fruit maturation under contrasting growing climates

ABSTRACTThis study employs a quantitative metabolomics approach to study the dynamics of primary metabolites in ‘Kinnow’ mandarin as influenced by maturation and ripening under contrasting growing climates. Fruit samples were harvested at regular intervals during the advanced stages of maturation and ripening from two contrasting growing locations, one under subtropical–arid (STA) climate and the other under subtropical–humid (STH) climate. Fruit juice samples were analysed using a targeted liquid chromatography-mass spectrometry (LC/MS) technique for identification and quantitation of primary metabolites such as sugars, organic acids and some vitamins. Significant (p < 0.05) differences in the metabolite concentrations were observed to be influenced by maturation stage and growing location. An increase in the concentration of sugars and B-complex vitamins was observed during the final stages of maturation approaching commercial maturity, whereas organic acids and vitamin C concentrations decreased. Fruit grown under STA climate exhibited a higher concentration of vitamin C and a lower concentration of organic acids than those grown under STH climate. The results suggest that fruit metabolism during the final stages of maturation and ripening under arid versus humid climatic conditions is a key determinant of the flavour and nutritional quality of ‘Kinnow’ mandarin.

Oocyte hydration in round goby Neogobius melanostomus from the Gulf of Gdańsk: another invasive strategy?

Summary The round goby (Neogobius melanostomus) is one of the most abundant non-native species in the Gulf of Gdansk that use various strategies to increase fecundity and achieve invasive success. Oocyte hydration appears to be a specific osmotic adaptation of round goby's gametes for successful fertilization and hatching. The aim of this study was to demonstrate details of oocyte final maturation and hydration during the spawning-capable phase in round goby and also compare various indices such as 17β-estradiol (E2), hepatosomatic index (HSI) and gonadosomatic index (GSI) among oocyte final maturation stages. E2 fluctuated according to the development of oocyte batches. HSI did not show significant differences throughout the oocyte maturation. GSI was an inaccurate estimator of oocyte maturation in round goby. This study, for the first time, shows that hydration does accompany oocyte final maturation in this species. These findings are important to the cognition of biology and the adaptive strategies of this invasive species to environmental factors and can be essential components of the protection and preservation of native living resources in the Gulf of Gdansk.

The Effect of Wnt Pathway Modulators on Human iPSC-Derived Pancreatic Beta Cell Maturation.

Current published protocols for targeted differentiation of human stem cells toward pancreatic β-cells fail to deliver sufficiently mature cells with functional properties comparable to human islet β-cells. We aimed to assess whether Wnt-modulation could promote the final protocol stages of β-cell maturation, building our hypothesis on our previous findings of Wnt activation in immature hiPSC-derived stage 7 (S7) cells compared to adult human islets and with recent data reporting a link between Wnt/PCP and in vitro β-cell maturation. In this study, we stimulated canonical and non-canonical Wnt signaling in hiPSC-derived S7 cells using syntetic proteins including WNT3A, WNT4, WNT5A and WNT5B, and we inhibited endogenous Wnt signaling with the Tankyrase inhibitor G007-LK (TKi). Whereas neither canonical nor non-canonical Wnt stimulation alone was able to mature hiPSC-derived S7 cells, WNT-inhibition with TKi increased the fraction of monohormonal cells and global proteomics of TKi-treated S7 cells showed a proteomic signature more similar to adult human islets, suggesting that inhibition of endogenous Wnt contributes toward final β-cell maturation.

Origin of Hematopoietic Stem Cells in Embryonic Development

Bone marrow is the main hematopoietic organ of mature mammals. Hematopoietic stem cells (HSCs) are maintained in it throughout an organism’s whole life; it also regulates the production of mature blood cells. Adult HSCs are characterized by polypotency, self-renewal, and a specific phenotype, as well as by the ability to direct migration to the hematopoietic organs. The ability of the HSCs to restore hematopoiesis after transplantation into another organism is the main functional criterion for their existence in tissue. However, HSCs, which function in postnatal ontogenesis and possess these properties, represent the final stage of maturation of their precursors (pre-HSCs), which arise in prenatal development. In embryogenesis, hematopoiesis occurs in several transitional blood-forming organs: the yolk sac, aorta-gonad-mesonephros (AGM), placenta, and liver. Despite the long history of research on hematopoietic system ontogenesis, the anatomical site of the origination of the first pre-HSCs, which give rise to the definitive HSC line, is still not entirely clear. The review summarizes modern concepts of the features of the hematopoietic cells formed in the yolk sac, AGM, and placenta, and their contribution to embryonic liver colonization and in definitive hematopoiesis. Further study of the mechanisms of HSC formation in embryogenesis is of undoubted importance, not only for an understanding of the fundamental aspects of hematopoietic system functioning but also for improvement in treatment methods for hematological diseases.

Effects of follicular fluid of preovulatory follicles of repeat breeder dairy cows with subclinical endometritis on oocyte developmental competence

The aims of the present study were to determine the concentrations of lipopolysaccharide (LPS), hormonal progesterone, estradiol-17β, insulin growth factor (IGF-1) and magnesium in the serum and the preovulatory follicle follicular fluid (FF) in repeat breeder (RB) cows without (nSCE) or with subclinical endometritis (SCE), and further to examine the effects of this FF on developmental competence of cattle oocytes. In Experiment 1, 13 of 23 clinically healthy Holstein RB cows were identified (uterine PMNs) to have SCE. The cows were estrous synchronized, and 6–12 h after detection of standing estrus, FF and blood of the preovulatory follicles were collected. The mean (±SD) LPS (862.3 ± 148.1 compared with 1063.4 ± 262.8 EU/ml, P = 0.04) and estradiol-17β (188.9 ± 15.8 compared with 162.0 ± 31.5 ng/ml, P = 0.02) concentrations of FF was different between nSCE and SCE cows. In Experiment 2, FF of RB cows with relatively lesser (nSCE, n = 4) and greater (SCE, n = 4) percentages of uterine PMNs was separately added to the oocyte maturation medium for in vitro embryo production. Addition of FF from SCE cows to the oocyte maturation medium resulted in a lesser rate of development to the blastocyst stage than that of the nSCE cows (21.9 ± 1.8 compared with 27.8 ± 2.5%, P < 0.05). Results of the present study indicate greater FF LPS concentration may result in a lesser quality microenvironment milieu for the final stages of oocyte maturation in RB dairy cows with subclinical endometritis. In addition, supplementation of oocyte maturation medium with FF of preovulatory follicles from RB cows with subclinical endometritis resulted in a lesser potential of in vitro oocyte developmental competence.

Beyond the NEC: Modulation of Herpes Simplex Virus Nuclear Egress by Viral and Cellular Components

Herpesvirus virions are complex molecular machines that, like most viruses, must be stable in the extracellular environment yet disassemble readily upon receipt of the appropriate molecular cues. While we have learned much about the assembly and composition of these fascinating particles, many central questions remain unanswered. This review focuses on an early stage in herpesvirus assembly called nuclear egress whereby viral DNA genomes packaged within an icosahedral protein capsid are translocated from the nucleoplasm across both the inner and outer nuclear membranes to reach the cytoplasm where the final stages of virion maturation take place. Here, I provide an overview of our current understanding of herpes simplex virus nuclear egress, the hurdles that the virus must overcome during nuclear egress, and discuss the literature pointing to the regulation and facilitation of this process by viral and cellular components.

SUPPLEMENTATION OF L-CARNITINE IN DIETS FOR SILVER CATFISH AND ITS EFFECTS ON REPRODUCTIVE ASPECTS

The objective of the present study was to assess the reproductive performance of silver catfish Rhamdia quelen fed with diets supplemented with growing levels of L-carnitine. 288 juveniles with average initial weight of 94.36 ± 3.76g, fed during 126 days with diets containing 400, 800, 1200, 1600 and 2000 mg. kg-1 of L-carnitine and one control diet. Females fed with diets supplemented with 400 mg. kg-1 of L-carnitine presented increase in fertilization and fertility, reduction in levels of triglycerides and plasma calcium when fed with diets supplemented with 1600 mg. kg-1 of L-carnitine. Males fed with diets supplemented with 1600 mg. kg-1 of L-carnitine presented increase in spermatozoa and improvement in semen quality. Structural changes were observed in the development of germ cells in females and males, even though all fish were in final stages of maturation in all treatments. The offspring from breeding animals fed with 400 mg. kg-1 of L-carnitine presented better potentials of development during the embryonic process. Therefore, the L-carnitine supplementation in diets stimulates the gonad development of breeding animals and also speeds up the initial development of silver catfish offspring.

Atypical septate junctions maintain the somatic enclosure around maturing spermatids and prevent premature sperm release in Drosophila testis.

ABSTRACTTight junctions prevent paracellular flow and maintain cell polarity in an epithelium. These junctions are also required for maintaining the blood-testis barrier, which is essential for sperm differentiation. Septate junctions in insects are orthologous to the tight junctions. In Drosophila testis, major septate junction components co-localize at the interface of germline and somatic cells initially, and then condense between the two somatic cells in a cyst after germline meiosis. Their localization is extensively remodeled in subsequent stages. We find that characteristic septate junctions are formed between the somatic cyst cells at the elongated spermatid stage. Consistent with previous reports, knockdown of essential junctional components – Discs-large-1 and Neurexin-IV – during the early stages disrupted sperm differentiation beyond the spermatocyte stage. Knockdown of these proteins during the final stages of spermatid maturation caused premature release of spermatids inside the testes, resulting in partial loss of male fertility. These results indicate the importance of maintaining the integrity of the somatic enclosure during spermatid coiling and release in Drosophila testis. It also highlights the functional similarity with the tight junction proteins during mammalian spermatogenesis.This article has an associated First Person interview with the first author of the paper.

Uncommon presentation of a huge intrathoracic ganglioneuroma in an 8-year-old child: a rare case report

Ganglioneuroma (GN) is a rare benign neurogenic tumor which arises from the neural crest cells that represent the final maturation stage of neuroblasts. GN is commonly encountered in adolescent or young adult. Until now, only a few cases of intrathoracic GN were reported in pediatric patients, especially in below 10 years of age. We report here an 8-year-old boy, who first presented to primary care with symptoms of upper respiratory tract infection. Radiologic investigations revealed a giant posterior mediastinal mass extending from T4 to T9 vertebrae without evidence of vascular invasion or infiltration into adjacent structures. The patient subsequently underwent left thoracotomy and resection of mass under general anesthesia. He was discharged 3 days after operation without complication. Histopathologic examination confirmed the diagnosis of thoracic GN. Grades of the neuroblastic differentiation increase with the median age at diagnosis. Although GNs are usually benign, they can grow aggressively and cause compression to an adjacent structure. Therefore, surgical resection is the only treatment. Debulking of tumor provides an alternative solution, especially when vital structures are involved. Complete resection remains the gold standard treatment for GN. However, in cases of incomplete resection, all residual tumors require regular clinical and radiodiagnostic follow-up.

Mast Cells as Key Players in Allergy and Inflammation.

Mast cells (MCs) are a key structural and functional component of both the innate and the adaptive immune systems. They are involved in many different processes, but play a major role in the response to infections and in inflammatory reactions. In addition, MCs are the main effector cells in allergy. MC biology is far more complex than initially believed. Thus, MCs may act directly or indirectly against pathogens and show a wide variety of membrane receptors with the ability to activate cells in response to various stimuli. Depending on where MCs complete the final stages of maturation, the composition of their cytoplasmic granules may vary considerably, and the clinical symptoms associated with tissue MC activation and degranulation may be also different. MCs are activated by complex signalling pathways characterized by multimolecular activating and inhibitory interactions. This article provides a comprehensive overview of MC biology, focusing predominantly on mechanisms of MC activation and the role of MCs in the pathogenesis of allergic diseases.

The Influence of Pentraxin 3 on the Ovarian Function and Its Impact on Fertility.

Follicular development is a highly coordinated process that in humans takes more than 6 months. Pituitary gonadotropins and a variety of locally produced growth factors and cytokines are involved in determining a precise sequence of changes in cell metabolism, proliferation, vascularization, and matrix remodeling in order to obtain a follicle with full ovulatory and steroidogenic capability. A low-grade inflammation can alter such processes leading to premature arrest of follicular growth and female reproductive failure. On the other hand, factors that are involved in inflammatory response as well as in innate immunity are physiologically upregulated in the follicle at the final stage of maturation and play an essential role in ovulation and fertilization. The generation of pentraxin 3 (PTX3) deficient mice provided the first evidence that this humoral pattern recognition molecule of the innate immunity has a non-redundant role in female fertility. The expression, localization, and molecular interactions of PTX3 in the periovulatory follicle have been extensively studied in the last 10 years. In this review, we summarize findings demonstrating that PTX3 is synthesized before ovulation by cells surrounding the oocyte and actively participates in the organization of the hyaluronan-rich provisional matrix required for successful fertilization. Data in humans tend to confirm these findings, indicating PTX3 as a biomarker of oocyte quality. Moreover, we discuss the emerging evidence that in humans altered PTX3 systemic levels, determined by genetic variations and/or low-grade chronic inflammation, can also impact the growth and development of the follicle and affect the incidence of ovarian disorders.

Sex hormone binding globulin during an annual reproductive cycle in the hepatopancreas and ovary of pejerrey (Odontesthes bonariensis)

In the present study, we determined the hepatopancratic shbg transcript abundance and ovarian immunoreactive Shbg (ir-Shbg) localization in pejerrey females at different gonadal stages during an annual ovarian cycle. In the hepatopancreas, shbg expression remains was constant in pre-vitellogenic stages, decreased at final vitellogenesis to increase again in final maturation and atretic stages to previous levels at post-vitellogenic stages. Related to this, also we found a negative significant relation between sex steroid and shbg expression. On the other hand, in the ovary we found ir-Shbg inside of cortical alveoli, from previtellogenic stages to final maturation. This localization of Shbg in a teleost fish ovary suggests a new role for Shbg in oocytes, that may also extend the oocyte fertilization/development process.

Human Dental Pulp Stem Cells and Gingival Mesenchymal Stem Cells Display Action Potential Capacity In Vitro after Neuronogenic Differentiation.

The potential of human mesenchymal stromal/stem cells (MSCs) including oral stem cells (OSCs) as a cell source to derive functional neurons has been inconclusive. Here we tested a number of human OSCs for their neurogenic potential compared to non-OSCs and employed various neurogenic induction methods. OSCs including dental pulp stem cells (DPSCs), gingiva-derived mesenchymal stem cells (GMSCs), stem cells from apical papilla and non-OSCs including bone marrow MSCs (BMMSCs), foreskin fibroblasts and dermal fibroblasts using non-neurosphere-mediated or neurosphere-mediated methods to guide them toward neuronal lineages. Cells were subjected to RT-qPCR, immunocytofluorescence to detect the expression of neurogenic genes or electrophysiological analysis at final stage of maturation. We found that induced DPSCs and GMSCs overall appeared to be more neurogenic compared to other cells either morphologically or levels of neurogenic gene expression. Nonetheless, of all the neural induction methods employed, only one neurosphere-mediated method yielded electrophysiological properties of functional neurons. Under this method, cells expressed increased neural stem cell markers, nestin and SOX1, in the first phase of differentiation. Neuronal-like cells expressed βIII-tubulin, CNPase, GFAP, MAP-2, NFM, pan-Nav, GAD67, Nav1.6, NF1, NSE, PSD95, and synapsin after the second phase of differentiation to maturity. Electrophysiological experiments revealed that 8.3% of DPSC-derived neuronal cells and 21.2% of GMSC-derived neuronal cells displayed action potential, although no spontaneous excitatory/inhibitory postsynaptic action potential was observed. We conclude that DPSCs and GMSCs have the potential to become neuronal cells in vitro, therefore, these cells may be used as a source for neural regeneration.

Variants in exons 5 and 6 of ACTB cause syndromic thrombocytopenia

Germline mutations in the ubiquitously expressed ACTB, which encodes β-cytoplasmic actin (CYA), are almost exclusively associated with Baraitser-Winter Cerebrofrontofacial syndrome (BWCFF). Here, we report six patients with previously undescribed heterozygous variants clustered in the 3′-coding region of ACTB. Patients present with clinical features distinct from BWCFF, including mild developmental disability, microcephaly, and thrombocytopenia with platelet anisotropy. Using patient-derived fibroblasts, we demonstrate cohort specific changes to β-CYA filament populations, which include the enhanced recruitment of thrombocytopenia-associated actin binding proteins (ABPs). These perturbed interactions are supported by in silico modeling and are validated in disease-relevant thrombocytes. Co-examination of actin and microtubule cytoskeleton constituents in patient-derived megakaryocytes and thrombocytes indicates that these β-CYA mutations inhibit the final stages of platelet maturation by compromising microtubule organization. Our results define an ACTB-associated clinical syndrome with a distinct genotype-phenotype correlation and delineate molecular mechanisms underlying thrombocytopenia in this patient cohort.