Cortical Stem Cells Research Articles

What Makes Us Human: Insights from the Evolution and Development of the Human Neocortex.

"What makes us human?" is a central question of many research fields, notably anthropology. In this review, we focus on the development of the human neocortex, the part of the brain with a key role in cognition, to gain neurobiological insight toward answering this question. We first discuss cortical stem and progenitor cells and human-specific genes that affect their behavior. We thus aim to understand the molecular foundation of the expansion of the neocortex that occurred in the course of human evolution, as this expansion is generally thought to provide a basis for our unique cognitive abilities. We then review the emerging evidence pointing to differences in the development of the neocortex between present-day humans and Neanderthals, our closest relatives. Finally, we discuss human-specific genes that have been implicated in neuronal circuitry and offer a perspective for future studies addressing the question of what makes us human.

Context-dependent regulation of Notch signaling in glial development and tumorigenesis.

In the mammalian brain, Notch signaling maintains the cortical stem cell pool and regulates the glial cell fate choice and differentiation. However, the function of Notch in regulating glial development and its involvement in tumorigenesis have not been well understood. Here, we show that Notch inactivation by genetic deletion of Rbpj in stem cells decreases astrocytes but increases oligodendrocytes with altered internal states. Inhibiting Notch in glial progenitors does not affect cell generation but instead accelerates the growth of Notch-deprived oligodendrocyte progenitor cells (OPCs) and OPC-related glioma. We also identified a cross-talk between oligodendrocytes and astrocytes, with premyelinating oligodendrocytes secreting BMP4, which is repressed by Notch, to up-regulate GFAP expression in adjacent astrocytes. Moreover, Notch inactivation in stem cells causes a glioma subtype shift from astroglia-associated to OPC-correlated patterns and vice versa. Our study reveals Notch's context-dependent function, promoting astrocytes and astroglia-associated glioma in stem cells and repressing OPCs and related glioma in glial progenitors.

An In Vivo Platform for Rebuilding Functional Neocortical Tissue.

Recent progress in cortical stem cell transplantation has demonstrated its potential to repair the brain. However, current transplant models have yet to demonstrate that the circuitry of transplant-derived neurons can encode useful function to the host. This is likely due to missing cell types within the grafts, abnormal proportions of cell types, abnormal cytoarchitecture, and inefficient vascularization. Here, we devised a transplant platform for testing neocortical tissue prototypes. Dissociated mouse embryonic telencephalic cells in a liquid scaffold were transplanted into aspiration-lesioned adult mouse cortices. The donor neuronal precursors differentiated into upper and deep layer neurons that exhibited synaptic puncta, projected outside of the graft to appropriate brain areas, became electrophysiologically active within one month post-transplant, and responded to visual stimuli. Interneurons and oligodendrocytes were present at normal densities in grafts. Grafts became fully vascularized by one week post-transplant and vessels in grafts were perfused with blood. With this paradigm, we could also organize cells into layers. Overall, we have provided proof of a concept for an in vivo platform that can be used for developing and testing neocortical-like tissue prototypes.

How mechanisms of stem cell polarity shape the human cerebral cortex.

Apical-basal progenitor cell polarity establishes key features of the radial and laminar architecture of the developing human cortex. The unique diversity of cortical stem cell populations and an expansion of progenitor population size in the human cortex have been mirrored by an increase in the complexity of cellular processes that regulate stem cell morphology and behaviour, including their polarity. The study of human cells in primary tissue samples and human stem cell-derived model systems (such as cortical organoids) has provided insight into these processes, revealing that protein complexes regulate progenitor polarity by controlling cell membrane adherence within appropriate cortical niches and are themselves regulated by cytoskeletal proteins, signalling molecules and receptors, and cellular organelles. Studies exploring how cortical stem cell polarity is established and maintained are key for understanding the features of human brain development and have implications for neurological dysfunction.

Abstract 204: Notch2NL is a novel regulator of radiation sensitivity in non-small cell lung cancer and a positive regulator of MYC activity

Abstract Background: A gain-of-function genetic screen identified NOTCH2 N-terminal like (NOTCH2NL) as a candidate novel gene involved in radiation resistance in non-small cell lung cancer (NSCLC). NOTCH2NL is located on chromosome position 1q21.1 which was incorrectly aligned in previous builds of the reference human genome sequence. NOTCH2NL was derived from a segmental duplication event in NOTCH2 during the evolutionary divergence between humans and non-human primates, forming three Notch2NL proteins: Notch2NL-A, Notch2NL-B and Notch2NL-C. Consequently, the only species whose genomes contain NOTCH2NL are modern and ancient humans. Notch2NL-B is a secreted protein and has been shown to promote Notch signalling in cortical stem cell progenitors via inhibition of inhibitory cis-DLL1/NOTCH signalling. Methods: Stable overexpression of NOTCH2NLB and secreted Notch2NL-B protein were validated by RT-qPCR and Western Blotting. The effects of NOTCH2NLB overexpression; transfer of Notch2NL-B protein from conditioned media onto wild-type cell lines; and siRNA knockdown of NOTCH1-4 transmembrane receptors on Notch signalling were quantified by a Notch luciferase reporter assay, RT-qPCR of classical NOTCH target genes and cancer stem cell marker (CSC) and RNA-Seq. Clonogenic cell survival assays determined the impact of NOTCH2NLB overexpression on radiation response. Cell proliferation was measured using the IncuCyte Zoom system. shRNA stable cell lines to knockdown NOTCH2NL were generated to mitigate the NOTCH2NLB overexpression phenotype we observed. RNA-Seq data from the first 100 patients from the TRACERx lung cancer study was used to quantify hallmark gene expression signatures and NOTCH2NLB expression. Results: NOTCH2NLB overexpression upregulates CSL-dependent Notch reporter luciferase activity; confers a radiation-resistant phenotype and impacts proliferation rate in NSCLC cell lines. Furthermore, we demonstrate that NOTCH2NLB overexpression and secreted Notch2NL-B protein transfer increase expression of classical NOTCH target genes (c-MYC, HES1 and HEY1) and CSC marker, ALDH1A1. Analysis of RNA-Seq data from the Lung TRACERx cohort study shows that NOTCH2NL is commonly expressed in NSCLC. NOTCH2NL expression positively correlates with notch hallmark gene expression signature in squamous (R=0.59, p=5.6e-9) and non-squamous carcinoma (R=0.51, p=4.7e-10). Interestingly, NOTCH2NL expression also positively correlated with the myc hallmark gene expression signature in squamous cell carcinoma (R=0.45, p=2.4e-5). Conclusions: Notch2NL is a novel determinant of radiation therapy response and impacts canonical Notch signalling in lung cancer. It is commonly expressed in NSCLC and can act in a paracrine manner as a secreted protein. Notch2NL, through canonical notch signalling, can regulate MYC expression and increase activity in the MYC pathway. Citation Format: Simranpreet Kaur Summan, Barbara Chow, Rhona Millar, Su Kit Chew, Philip East, Robert Hynds, Carlos Martinez Ruiz, Eva Gronroos, Mariam Jamal-Hanjani, Kevin Litchfield, Nicholas McGranahan, Nnennaya Kanu, Charles Swanton, Crispin Thomas Hiley. Notch2NL is a novel regulator of radiation sensitivity in non-small cell lung cancer and a positive regulator of MYC activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 204.

Protective Mechanism of Proprotein Convertase Subtilisin-Like Kexin Type 9 Inhibitor on Rats with Middle Cerebral Artery Occlusion-Induced Cerebral Ischemic Infarction.

The aim of this study was to research the mechanism of proprotein convertase subtilisin-like kexin type 9 (PCSK9) inhibitor in neural protective effect on rat cerebral ischemic reperfusion injury (I/RI). The transient middle cerebral artery occlusion (tMCAO) model of rats was prepared by the suture method, and PCSK9 inhibitor was injected intraperitoneally immediately after I/R. The rats were scored for neurological deficits and the cerebral infarction volume was measured. The brain tissues were collected and western blot (WB) was used to detect the expression of PCSK9. The rat cortical neural stem cells were treated with oxygen glucose deprivation (OGD) to establish a cell model of ischemia/reperfusion. WB was used to detect the expression of PCSK9 and the apoptosis-related pathway proteins. After interfering with the expression of PCSK9 siRNA, the cell viability (cell counting kit-8 assay) and apoptosis (TUNEL staining, Annexin V/PI method) were detected, and the cell proliferation was detected by EdU staining and flow cytometry. The expression of PCSK9 in the brain tissue of the MCAO group was dramatically increased. PCSK9 inhibitor can improve neurobehavioral scores and reduce apoptosis and infarct volume. An OGD model of neural stem cells in vitro was constructed. Inhibiting PCSK9 with si-PCSK9 can increase cell viability, promote cell proliferation, and also reduce cell apoptosis. Inhibition of PCSK9 can decrease the cerebral infarct volume in rats with cerebral I/RI and improve the neural function. Mechanically, inhibition of PCSK9 can lead to the decrease of nerve cell apoptosis and promotion of cell proliferation.

Melatonin attenuates radiation-induced cortical bone-derived stem cells injury and enhances bone repair in postradiation femoral defect model

BackgroundThe healing of bone defects can be challenging for clinicians to manage, especially after exposure to ionizing radiation. In this regard, radiation therapy and accidental exposure to gamma (γ)-ray radiation have been shown to inhibit bone formation and increase the risk of fractures. Cortical bone-derived stem cells (CBSCs) are reportedly essential for osteogenic lineages, bone maintenance and repair. This study aimed to investigate the effects of melatonin on postradiation CBSCs and bone defect healing.MethodsCBSCs were extracted from C57BL/6 mice and were identified by flow cytometry. Then CBSCs were subjected to 6 Gy γ-ray radiation followed by treatment with various concentrations of melatonin. The effects of exogenous melatonin on the self-renewal and osteogenic capacity of postradiation CBSCs in vitro were analyzed. The underlying mechanisms involved in genomic stability, apoptosis and oxidative stress-related signaling were further analyzed by Western blotting, flow cytometry and immunofluorescence assays. Moreover, postradiation femoral defect models were established and treated with Matrigel and melatonin. The effects of melatonin on postradiation bone healing in vivo were evaluated by micro-CT and pathological analysis.ResultsThe decrease in radiation-induced self-renewal and osteogenic capacity were partially reversed in postradiation CBSCs treated with melatonin (P < 0.05). Melatonin maintained genomic stability, reduced postradiation CBSC apoptosis and intracellular oxidative stress, and enhanced expression of antioxidant-related enzymes (P < 0.05). Western blotting validated the anti-inflammatory effects of melatonin by downregulating interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) levels via the extracellular regulated kinase (ERK)/nuclear factor erythroid 2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) signaling pathway. Melatonin was also found to exhibit antioxidant effects via NRF2 signaling. In vivo experiments demonstrated that the newly formed bone in the melatonin plus Matrigel group had higher trabecular bone volume per tissue volume (BV/TV) and bone mineral density values with lower IL-6 and TNF-α levels than in the irradiation and the Matrigel groups (P < 0.05).ConclusionThis study suggested that melatonin could protect CBSCs against γ-ray radiation and assist in the healing of postradiation bone defects.

Leaf and stem anatomical traits of Muntingia calabura L. (Muntingiaceae) emphasizing the production sites of bioactive compounds

Muntingia calabura L. is a neotropical Muntingiaceae tree with medicinal properties and economic interest. Extracts from its leaves and stems are rich in carbohydrates, volatiles and phenolic compounds with proved anti-tumor, antinociceptive and thermogenic properties. However, little is known about the structural features of leaves and stem and where these compounds are produced. In addition to the glandular trichomes registered in the literature, we hypothesized that other morphotypes of secretory cells and tissues could be present in M. calabura. In this paper, we analysed the leaf and stem anatomical traits of M. calabura to assess the production sites of bioactive compounds. Leaf and stem samples were collected from plants living in an urban area in Botucatu city, southeastern Brazil, and submitted to standard methods for anatomical and histochemical assessments. The leafblade was hypoestomatic, heterobaric and exhibited dorsiventral mesophyll. The petiole had parenchyma cortex, central vascular system and parenchyma pith. The stem presented parenchyma cortex, peripheral vascular system and parenchyma pith. Druses of calcium occurred mainly in the inner cortex of stem, petiole and midrib. Abundant glandular and non-glandular trichomes characterized the epidermis of leaves and stem. Phenolic compounds and mucilage were detected in the glandular trichome head cells in all the analysed organs, in addition to the cortical cells of midrib, petiole and stem. Voluminous bi-compartmentalized cells containing phenolic compounds and mucilage characterized the epidermis in the adaxial surface of the leaf blade. These cells were a remarkable feature that deserve further investigation. Our results shed light to the poorly known secretory system of M. calabura and proved the occurrence of different morphotypes of secretory cells and tissues in its leaves and stem. The obtained data can guide more studies with ecological, structural and taxonomic approaches, and for researches on drug bioprospecting from this species.

Stem Cell Treatment for Ischemic Stroke Recovery.

The role of cellular transplantation to promote functional recovery after stroke has been evaluated over the last two decades. Preclinical studies first established the potential for cultured neuronal cells derived from a teratocarcinoma cell line to be tested for safety and efficacy in the treatment of human stroke. In animal models of stroke that caused reproducible learning and motor deficits, injection of neuronal cells resulted in a return of learning behavior, retention time, and motor function. Clinical trials followed. Additional work with cells derived from a bone marrow neuroprogenitor line, fetal cortical stem cells, and other cell sources showed promise in preclinical studies and then these cells were tested in clinical studies. This report reviews the different biological repair approaches using cell implants, discusses clinical trial design and surgical methods, and the current state of research.

Mitochondrial dynamics in postmitotic cells regulate neurogenesis.

The conversion of neural stem cells into neurons is associated with the remodeling of organelles, but whether and how this is causally linked to fate change is poorly understood. We examined and manipulated mitochondrial dynamics during mouse and human cortical neurogenesis. We reveal that shortly after cortical stem cells have divided, daughter cells destined to self-renew undergo mitochondrial fusion, whereas those that retain high levels of mitochondria fission become neurons. Increased mitochondria fission promotes neuronal fate, whereas induction of mitochondria fusion after mitosis redirects daughter cells toward self-renewal. This occurs during a restricted time window that is doubled in human cells, in line with their increased self-renewal capacity. Our data reveal a postmitotic period of fate plasticity in which mitochondrial dynamics are linked with cell fate.

Abstract 342: Cortical Bone Stem Cell Therapy Alters Macrophage Phenotype and Reduces Cardiac Cell Death After Myocardial Infarction

Acute injuries to the heart, like myocardial infarction (MI), contribute to the development and pathology of heart failure (HF). Reperfusion of the ischemic heart greatly increases survival but results in reperfusion injury that can account for up to 50% of the final infarct size. The inflammatory response to MI-induced myocardial injury is thought to be responsible for the propagation of reperfusion injury into the infarct border zone, expanding myocardial damage. We have previously shown in a swine model of MI that intramyocardial injections of cortical bone-derived stem cells (CBSCs) into the infarct border zone has no acute cardioprotective effect but reduces scar size by half and prevents the decline of ejection fraction and LV dilation 3 months after MI. Our new preliminary data show that CBSCs have potent immunoregulatory capabilities. Therefore, we hypothesize that CBSC treatment has an effect on the immune response to MI that improves the wound healing response to myocardial injury and mitigates LV remodeling and infarct size 3 months later. To test this hypothesis, we characterized the effects of CBSC paracrine factors on macrophages in vitro and found that CBSC-treated macrophages express higher levels of CD206, produce more IL-1RA and IL-10, and phagocytose apoptotic myocytes more efficiently. In addition, macrophages were increased in CBSC-treated swine hearts 7 days after MI compared to controls with a corresponding increase in IL-1RA and TIMP-2. Apoptosis was decreased overall and in macrophages specifically in CBSC-treated animals. From these data we conclude CBSCs may exert an acute pro-reparative effect on the immune response after MI, reducing reperfusion injury and adverse remodeling resulting in improved functional outcomes at later time points.

OR19-02 Luteinizing Hormone/Human Chorionic Gonadotropin Receptor Protein Expression in Adrenocortical Progenitor Cells, Aldosterone Producing Cell Clusters and Adrenal Adenomas Derived from Postmenopausal Women

Objective/BackgroundAdrenal pathologies are more common in women than men. Embryologically the adrenals and gonads develop from the adrenogenital ridge with differential migration and differentiation. We hypothesized that in adult females there are adrenocortical progenitor cells that express the LH/hCG-R and proliferate in response to elevated LH. Indeed, several case reports demonstrated LH/hCG-R expression in adrenal secretory tumors in postmenopausal and pregnant females. In aging adults, nests of cells known as aldosterone-producing cell clusters (APCCs) that may be precursors to aldosterone producing adenomas are frequently detected. We retrospectively studied the immunohistochemical expression of LH/hCG-R in normal adrenals, adrenal adenomas and APCCs in archival specimens derived from post-menopausal women.MethodsArchival specimens from adrenal adenomas derived from 23 women >55 years of age were examined. Clinical data was obtained in a blinded fashion and hormonal data was available in 9/23 cases; 6/9 were secreting cortisol and 3/9 adenomas were secreting aldosterone. In addition, 6 samples derived from a repository of normal adrenal tissues from deceased kidney donors (1 male, and 5 postmenopausal females) were studied. All specimens were immunostained for LH/hCG-R and the adrenal stem cell marker DLK1 that facilitates the maintenance of an undifferentiated phenotype. The normal adrenal tissues were also stained for aldosterone synthase (CYP11B2) to detect APCCs. The slides were reviewed and graded by a pathologist in a blinded fashion.ResultsExpression of LH/hCG-R was demonstrated in both normal and adenomatous tissues in all 23 specimens. The staining in adenomas was heterogeneous, with clusters of densely stained LH/hCG-R positive cells in all specimens. There were less densely stained clusters in normal adjacent adrenocortical tissue that was most prominent in the subcapsular, zona glomerulosa region, an area where the putative adrenal cortical stem cells are found as well as the zona reticularis. Double staining for the stem cell marker DLK1 and LH/hCG-R confirmed that these cells represent adrenocortical progenitor cells. CYP 11B2 immunohistochemistry of normal adrenals demonstrated cell foci dipping from the capsule into the zona fasciculata classified as APCCs that co-expressed cytoplasmic LH/hCGR.ConclusionAdrenal adenomas and APCCs derived from postmenopausal women exhibited heterogeneous but strong immunohistochemical expression of LH/hCG-R in all samples. Interestingly, DLK1-positive adrenocortical stem cells in the subcapsular zone also expressed LH/hCG-R. These data may provide insights into the female predominance of adrenal pathologies, particularly in postmenopausal women with high LH levels. The LH/hCG-R may be a viable target for treatment of adrenal adenomas in postmenopausal women.

Sp2 regulates late neurogenic but not early expansive divisions of neural stem cells underlying population growth in the mouse cortex.

Cellular and molecular mechanisms underlying the switch from self-amplification of cortical stem cells to neuronal and glial generation are incompletely understood, despite their importance for neural development. Here, we have investigated the role of the transcription factor specificity protein 2 (Sp2) in expansive and neurogenic divisions of the developing cerebral cortex by combining conditional genetic deletion with the mosaic analysis with double markers (MADM) system in mice. We find that loss of Sp2 in progenitors undergoing neurogenic divisions results in prolonged mitosis due to extension of early mitotic stages. This disruption is correlated with depletion of the populations of upper layer neurons in the cortex. In contrast, early cortical neural stem cells proliferate and expand normally in the absence of Sp2. These results indicate a stage-specific requirement for Sp2 in neural stem and progenitor cells, and reveal mechanistic differences between the early expansive and later neurogenic periods of cortical development.This article has an associated 'The people behind the papers' interview.

Abstract 257: Cortical Bone Stem Cell-Derived Exosomes Alter Wound Healing Response in Cardiac Fibroblasts and Cardiac Endothelial Cells

Rationale: Post-myocardial infarction (MI) results in remodeling that leads to the development of heart failure. We have previously seen improvements in post-MI wound healing and scar formation as a result of administration of cortical bone stem cell derived exosomes (CBSC exosomes). Objectives: We sought to further elucidate the mechanism through which CBSC exosomes improved scar formation and altered wound healing through in vitro experimentation. We continued to explore the unique characteristics of CBSC exosomes in order to understand why they have anti-fibrotic effects. Methods and Results: We performed migration assays on cardiac endothelial cells and cardiac fibroblasts, and saw alterations in cell migration in both cell types upon treatment with mCBSC CM and mCBSC exosomes. We discovered that migration of CECs was reduced by 20% under the influence of mCBSC CM and mCBSC exosomes, but rescued when exosomes were removed from mCBSC CM. We discovered that the removal of exosomes from mCBSC CM slowed cardiac fibroblast migration by 40%. We saw a decrease in the quantity of pro-fibrotic mRNA in cardiac endothelial cells and cardiac fibroblasts after treatment with mCBSC exosomes. Adult cardiac endothelial cell mRNA levels of VEGFA, Col4A1, and VCAM were reduced nearly two fold from control untreated cell levels following mCBSC exosome treatment. Adult cardiac fibroblast Col1A1, Col3A1, Col4A1, and MMP2 mRNA levels were reduced one-fold from control untreated cell levels following mCBSC exosome treatment. Conclusions: Our findings show that the wound healing induced by CBSC treatment post-MI involves a mechanism altering the migration of endothelial cells, sustaining the migratory capabilities of fibroblasts, and decreasing the production of pro-fibrotic mRNA in cardiac fibroblasts and cardiac endothelial cells.

Abstract 707: Cortical-bone Stem Cell Therapy Alters the Inflammatory Response After Myocardial Infarction

Ischemic heart diseases like myocardial infarction are the largest contributors to cardiovascular disease world-wide. The resulting cardiac cell death impairs function of the heart and can lead to heart failure and death. Re-perfusion of the ischemic tissue is necessary but causes damage to the surrounding tissue by re-perfusion injury and initiates a sterile inflammatory response. Cortical bone stem cells (CBSCs) have been shown to increase pump function and decrease scar size in a large animal swine model of myocardial infarction. To explore the potential cause of these changes, we hypothesized that CBSCs were altering the inflammatory response after re-perfusion thereby changing the wound healing process. To test this, we performed serial immune cell analysis of the blood and tissue from Gottingen mini-swine that underwent 90 minutes of lateral anterior descending coronary artery ischemia followed by 7 days of re-perfusion to assess changes in immune cell recruitment and phenotype. Our findings indicate that CBSCs modify the cytokines released by immune cells in the infarct, decrease macrophage infiltration of the infarct 3 days after MI, and increase the recruitment of CD4+ T-cells to the infarct zone 7 days after MI. In addition, these changes reflect a pro-healing inflammatory environment. From this data, we conclude that CBSCs are influencing immune cell recruitment dynamics and phenotype, and these changes may contribute to the decreased scar size and increased pump function seen in CBSC-treated animals.

SAT-343 Immunohistochemical Expression of Luteinizing Hormone/Human Chorionic Gonadotropin Receptor (LH/hCG Receptor) in Adrenal Adenomas of Postmenopausal Women

Background: Increasingly, patients are found to have incidentally found adrenal adenomas on imaging performed for other purposes. Further biochemical work-up results in a diagnosis of mild hypercortisolism in as many as 30% of these patients. The incidence of adrenal adenomas and Cushing’s is higher in women than men with peak incidence ages 50-70. Elevated expression of gonadal markers such as LH/hCG receptor and the gonadal transcription factor GATA-4 has been well described in post-gonadectomy induced adrenal cortical neoplasia in neutered ferrets and certain mouse strains. The majority of these neoplasms of the adrenal cortex, which share a common embryologic progenitor with the gonads in the urogenital ridge, produce sex steroids and appear to differentiate towards a “gonadal” molecular profile. Isolated case reports have demonstrated LH/hCG receptor expression in adrenal secretory tumors in postmenopausal women and pregnant females. Taken together, there is strong evidence that a subset of adrenal tumors is driven by LH in postmenopausal women. Hypothesis: We hypothesized that adrenal adenomas producing secretory syndromes in postmenopausal women express LH/hCG receptor. Methods: 24 archival adrenal tissues from women >50 years of age with hypersecretory syndromes (primary aldosteronism and Cushing’s) were obtained with IRB approval. The samples were immunostained for expression of the LH/hCG receptor (monoclonal unconjugated LHR Antibody (LHCGR/1417), Novus Biologicals, CO) utilizing the Streptavidin–Biotin staining method. The slides were reviewed in a blinded fashion, and cytoplasmic, membrane, and nuclear expression of the markers were graded as no immunoreactivity, or 1+, 2+, or 3+ staining. Results: All 24 adenomas had demonstrated expression of LH/hCG receptor in both normal and adenomatous tissue. The staining in adenomas was heterogeneous, with clusters of densely stained LH/hCG receptor positive cells found selectively in areas of the adenomas. In addition, these clusters were found in normal adjacent adrenal tissue primarily in the subcapsular, zona glomerulosa region, an area where the putative adrenal cortical stem cells are found. Conclusion: LH/hCG receptor was found to be expressed in normal adrenocortical and adenomatous tissue in postmenopausal women, a population that has elevated serum LH levels. These findings may have clinical implications and provide a rationale for the treatment of secretory adrenal adenomas with GnRH agonists or antagonists in postmenopausal women.

Neuroprotective effects of creatine treatment on the differentiation of E14 rat cortical stem cells and expression of GABA-ergic neurons.

Along with the promotion of muscle growth and recovery, it has been observed that the supplement, creatine, also contains neuroprotective properties that aid in the developmental process of neuronal cells by increasing developmental competency in early differentiation. In clinical research, addiction treatments are beginning to introduce dietary supplements such as Creatine for patients in early recovery to improve recovery abstinence. Research has shown that supplemental Creatine induces higher cerebral phosphocreatine levels which could help with improved neuron connectivity. In order to examine the neuroprotective effects of Creatine, E14 rat cortical stem cells were differentiated and treated with different dosages of alcohol and creatine. Alcohol was used as a negative control in order to compare the differentiation and expression of GABAergic neurons associated with alcohol exposure and neuronal protection from Creative. We anticipate that Creatine acts as a neuroprotective agent in GABAergic neurons treated with alcohol

THE EFFECT OF VARIOUS ALCOHOL DOSAGES ON NEURAL VESICLE PACKAGING AND GABA RECEPTORS.

Abusive alcohol consumption has negative effects on proliferation, differentiation and migration of neuronal stem cell in both fetal development and adult neural plasticity. IN both cases, loss of neurons and astrocytes alters synaptic transmission in dopaminergic and GABAergic synapses within the limbic system. This is particularly evident within areas of the amygdala and nuclear accumbens. It is known that alcohol causes an imbalance between GABA and glutamate-NMDA transmission, and could cause altered levels of neurotransmitter reuptake or insensitivity to regular GABA levels. In this study, the effects of binge and chronic alcohol exposure to neuronal stem cell were observed. We examined synaptic vesicle packaging and GABAergic receptors on fully differentiated neurons. E14 Rat Cortical Stem Cells were grown, and differentiated, then exposed to alcohol in various doses. The expected results were to observe alterations in the density of synaptic vesicles, and an alteration in GABAergic receptor expression. These results would suggest that both chronic and binge drinking of alcohol causes alterations in synaptic transmission as well as neuron numbers.

NEUROPROTECTIVE EFFECTS OF CREATINE TREATMENT ON THE DIFFERENTIATION OF E14 RAT CORTISOL STEM CELLS AND EXPRESSION OF GABA-ERGIC NEURONS

Along with the promotion of muscle growth and recovery, it has been observed that the supplement, Creatine, also contains neuroprotective properties that aid in the developmental process of neuronal cells by increasing developmental competency in early differentiation. In clinical research, addiction treatments are beginning to introduce dietary supplements such as Creatine for patients in early recovery to improve recovery abstinence. Research has shown that supplemental Creatine induces higher cerebral phosphocreatine levels which could help with improved neuron connectivity. In order to examine the neuroprotective effects of Creatine, E14 rat cortical stem cells were differentiated and treated with different dosages of alcohol and creatine. Alcohol was used as a negative control in order to compare the differentiation and expression of GABAergic neurons associated with alcohol exposure and neuronal protection from Creative. We anticipate that Creatine acts as a neuroprotective agent in GABAergic neurons treated with alcohol.

The effect of various alcohol dosages on neural vesicle packaging and GABA receptor expression

Abusive alcohol consumption has negative effects on proliferation, differentiation and migration of neuronal stem cell in both fetal development and adult neural plasticity. IN both cases, loss of neurons and astrocytes alters synaptic transmission in dopaminergic and GABAergic synapses within the limbic system. This is particularly evident within areas of the amygdala and nuclear accumbens. It is known that alcohol causes an imbalance between GABA and glutamate-NMDA transmission, and could cause altered levels of neurotransmitter reuptake or insensitivity to regular GABA levels. In this study, the effects of binge and chronic alcohol exposure to neuronal stem cell were observed. We examined synaptic vesicle packaging and GABAergic receptors on fully differentiated neurons. E14 Rat Cortical Stem Cells were grown, and differentiated, then exposed to alcohol in various doses. The expected results were to observe alterations in the density of synaptic vesicles and an alteration in GABAergic receptor expression. These results would suggest that both chronic and binge drinking of alcohol causes alterations in synaptic transmission as well as neuron numbers.