Astrocyte Granules Research Articles

Oligodendroglial fatty acid metabolism as a central nervous system energy reserve

Brain function requires a constant supply of glucose. However, the brain has no known energy stores, except for glycogen granules in astrocytes. In the present study, we report that continuous oligodendroglial lipid metabolism provides an energy reserve in white matter tracts. In the isolated optic nerve from young adult mice of both sexes, oligodendrocytes survive glucose deprivation better than astrocytes. Under low glucose, both axonal ATP levels and action potentials become dependent on fatty acid β-oxidation. Importantly, ongoing oligodendroglial lipid degradation feeds rapidly into white matter energy metabolism. Although not supporting high-frequency spiking, fatty acid β-oxidation in mitochondria and oligodendroglial peroxisomes protects axons from conduction blocks when glucose is limiting. Disruption of the glucose transporter GLUT1 expression in oligodendrocytes of adult mice perturbs myelin homeostasis in vivo and causes gradual demyelination without behavioral signs. This further suggests that the imbalance of myelin synthesis and degradation can underlie myelin thinning in aging and disease.

Commentary: Granins, Secretory Granule Biogenesis, and Transport

Secretory granules are the hallmark of secretory cells that secrete granins, peptide hormones, and enzymes for various physiological functions in endocrine, exocrine, and neurons. The granin family of proteins are highly acidic and are the major components in dense core secretory granules, including those in chromaffin cells. Granins include chromogranin (Cg) A, B, and secretogranin (Sg) II which are processed to various derived peptides. In this 16th International Symposium of Chromaffin Cell Biology, a number of papers focusing on the mechanisms of biogenesis and transport of secretory granules in chromaffin cells, other endocrine cells and astrocytes were presented. As well, new physiological roles of granin-derived peptides were discussed. Maite Montero-Hadjadje (France) demonstrated that via its terminal domains, CgA induced granule biogenesis and generated a regulated secretory pathway in fibroblasts. These granules recruited cytoskeletal proteins, actin/myosin and microtubule motors to their surface and exhibited Ca sensitive transport to the plasma membrane for release. Peng Loh (USA) showed that a C-terminal CgA-derived peptide, serpinin, upon being released after stimulation of the cells, upregulated the transcription of a protease inhibitor, protease nexin-1 (PN-1) in a cAMP-PKA-sp1 dependent pathway. This in turn stabilized and augmented the level of granule proteins in the Golgi complex, leading to enhanced granule biogenesis. Ricardo Borges (Spain) using chromaffin cells from CgA knockout (KO), CgB KO and Cg double KO mice showed that the absence of granins led to changes in the accumulation and secretion of catecholamines. They concluded that these Cgs are highly efficient in facilitatingmonoamine and calcium accumulation and in the kinetics of exocytosis of dense core granules (for more details, see review by Borges group below). Seung Yoo (Korea) demonstrated the presence of inositol 1,4,5, triphosphate (IP3) sensitive Ca stores and three forms of IP3 receptors/channels in secretory granules in astrocytes, that presumably interacts with the Cgs to modulate Ca channels and Ca release as in chromaffin granules. Several papers were presented demonstrating the role of cytoskeletal network/proteins in granule transport to the plasma membrane for exocytosis. Luis Gutierrez (Spain) presented confocal microscopy and 3D reconstructed images revealing the complex F-actin structure at the cortex of chromaffin cells. He showed that this F-actin structure underwent reorganization during secretion to reposition the vesicles for exocytosis, (for more details, see their review below). Heidi de Wit (The Netherlands) recapped her extensive studies on the morphology of synaptic protein knockout cells at the electron microscopy level, suggesting that syntaxin, SNAP25 and synaptogamin form the minimal docking machinery. She also discussed the role of Munc 18 in reorganizing cortical F-actin for vesicle docking at the plasma membrane, prior to secretion, in chromaffin cells of embryonic mice. (See her review below). Frederic Meunier (Australia) described a novel phosphatidylinisitol (4,5) biphosphate pathway that controls actin-mediated mobilization and translocation of secretory vesicles to the plasma membrane. Small GTPases such as the Rho GTPase family are involved in actin remodeling during exocytosis. Stephane Gasman (France) showed in chromaffin cells and PC12 cells that Rho A, which is associated with secretory Y. P. Loh (*) Section on Cellular Neurobiology, Program on Developmental Neuroscience, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA e-mail: lohp@mail.nih.gov

Trafficking and Fusion of Neuropeptide Y-Containing Dense-Core Granules in Astrocytes

It is becoming clear that astrocytes are active participants in synaptic functioning and exhibit properties, such as the secretion of classical transmitters, previously thought to be exclusively neuronal. Whether these similarities extend to the release of neuropeptides, the other major class of transmitters, is less clear. Here we show that cortical astrocytes can synthesize both native and foreign neuropeptides and can secrete them in a stimulation-dependent manner. Reverse transcription-PCR and mass spectrometry indicate that cortical astrocytes contain neuropeptide Y (NPY), a widespread neuronal transmitter. Immunocytochemical studies reveal NPY-immunoreactive (IR) puncta that colocalize with markers of the regulated secretory pathway. These NPY-IR puncta are distinct from the synaptic-like vesicles that contain classical transmitters, and the two types of organelles are differentially distributed. After activation of metabotropic glutamate receptors and the release of calcium from intracellular stores, the NPY-IR puncta fuse with the cell membrane, and the peptide-containing dense cores are displayed. To determine whether peptide secretion subsequently occurred, exocytosis was monitored from astrocytes expressing NPY-red fluorescent protein (RFP). In live cells, after activation of glutamate receptors, the intensity of the NPY-RFP-labeled puncta declined in a step-like manner indicating a regulated release of the granular contents. Because NPY is a widespread and potent regulator of synaptic transmission, these results suggest that astrocytes could play a role in the peptidergic modulation of synaptic signaling in the CNS.

Very‐high‐dose α‐tocopherol supplementation increases blood pressure and causes possible adverse central nervous system effects in stroke‐prone spontaneously hypertensive rats

Tocopherols and tocotrienols constitute the vitamin E family. Although alpha-tocotrienol is the most neuroprotective form of vitamin E proved to be effective against stroke, alpha-tocopherol is the most abundant in nature and is used most often for disease prevention/treatment. A recent metaanalysis of human studies suggested that alpha-tocopherol supplementation increases all-cause mortality. Therefore, we investigated the effects of alpha-tocopherol ( approximately 44 mg/kg body weight; equivalent to 2,600 mg/human/day) on the central nervous system (CNS) of stroke-prone spontaneously hypertensive rats (SHRSP). SHRSP treated with high dose alpha-tocopherol had significantly higher blood pressure than untreated controls fed a basal diet that contained approximately 4 mg tocopherols/kg body weight, but neither group experienced a change in degree of lipid peroxidation in serum or CNS tissue. Biochemical/immunohistochemical analyses demonstrated that expressions of phosphorylated neurofilament H protein, glial fibrillary acidic protein and cathepsin D in the CNS tissue were significantly enhanced in alpha-tocopherol-supplemented rats, whereas expressions of SOD2 and Bcl-xL were diminished in response to alpha-tocopherol supplementation. Similarly, the frequency of cathepsin D-positive cells, corresponding mostly to microglial cells, was significantly increased in alpha-tocopherol-supplemented rats. Alpha-tocopherol supplementation also increased the number of lysosomes and lipofuscin granules in perikarya of both hippocampal pyramidal and Purkinje cells. Furthermore, alpha-tocopherol supplementation increased the frequency of glial filaments and lipofuscin granules in astrocytes and lysosomes in microglial cells that were frequently occupied with phagocytosed inclusion structures. The present results are the first to suggest that a very high dose of alpha-tocopherol supplementation increases blood pressure in SHRSP rats and influences the CNS tissue in a manner that seems adverse.

Regression stage senile plaques in the natural course of Alzheimer’s disease

Resolution process of cerebroparenchymal amyloid beta-protein (Abeta) deposition has become of increasing interest in the light of recent advance in the Abeta-vaccination therapy for Alzheimer's disease (AD). However, the neuropathological features of degraded and disappearing senile plaque remain poorly characterized, especially in the natural course of the disease. To clarify the natural removal processes of Abeta burden in the brain with AD, we devised a triple-step staining method: Bodian for dystrophic neurites, anti-glial fibrillary acidic protein for astrocytes, and anti-Abeta. We thus examined 24 autopsied AD brains. A novel form of senile plaques, termed 'remnant plaques', was identified. Remnant plaques were characterized by mesh-like astroglial fibrils within the entire plaque part, Abeta deposit debris exhibiting weak Abeta immunoreactivity, and only a few slender dystrophic neurites. In remnant plaques, amyloid burden was apparently decreased. The density of remnant plaques increased significantly with disease duration. Dual-labelling immunohistochemistry revealed many Abeta-immunoreactive granules in astrocytes and a modest number in microglia, both of which accumulated in senile plaques. We consider amyloid deposits of diffuse and neuritic plaques to be shredded by astrocytic processes from the marginal zone of plaques, and to gradually disintegrate into smaller compartments. Cerebroparenchymal Abeta deposits undergo degradation. After a long-standing resolution process, diffuse and neuritic plaques may finally proceed to remnant plaques. Astrocytes are actively engaged in the natural Abeta clearance mechanism in advanced stage AD brains, which may provide clues for developing new therapeutic strategies for AD.

Acceleration of ageing-related gliopathic changes and hippocampal dysfunction following intracerebroventricular infusion of cysteamine in adult rats

The sulphydryl agent, cysteamine, accelerates the ageing-related accumulation of peroxidase-positive (iron-rich) cytoplasmic inclusions in rat subcortical astroglia and induces their appearance in primary neuroglial cultures. In the present study, infusion of cysteamine into the lateral ventricle of young, adult rats (1 mg/day for three weeks followed by a one-month drug “washout” period) significantly increased numbers of peroxidase-positive astrocytic granules in the stratum oriens of the CA1 hippocampus relative to saline-infused controls. In contrast to the gliopathic changes, no evidence of neuronal or myelin damage was observed in the cysteamine-exposed rats. The cysteamine-treated animals exhibited significant impairment in spatial learning as determined using a three-panel runway task. The working memory deficits were more robust at the end of the drug washout period than immediately following cessation of the cysteamine infusion. Thus, the cysteamine-related memory deficits are of long duration and are not due to any acute neuroactive properties of the drug itself. Using hippocampal slices prepared after the drug washout period, we observed attenuated paired-pulse depression, with no significant effects on basal excitatory synaptic transmission or induction of long-term potentiation, in the cysteamine-infused animals relative to controls. We propose that, in cysteamine-treated rats and in the course of normal ageing, hippocampal dysfunction and associated cognitive deficits may be secondary to fundamental pathological processes originating within the astroglial compartment.

Identification of an astrocyte cell population from human brain that expresses perforin, a cytotoxic protein implicated in immune defense.

The brain is an immunoprivileged organ isolated from the peripheral immune system. However, it has been shown that resident cells, notably astrocytes and microglia, can express numerous innate immune molecules, providing the capacity to generate a local antipathogen system. Perforin is a cytolytic protein present in the granules of cytotoxic T lymphocytes and natural killer cells. Expression in cells other than those of the hemopoetic lineage has not been described. We report here that fetal astrocytes in culture (passages 2 to 15), astrocytoma, and adult astrocytes expressed perforin. Reverse transcriptase polymerase chain reaction followed by Southern blot was carried out using multiple specific primers and all cDNAs were cloned and sequenced. Human fetal astrocyte perforin cDNA sequence was approximately 100% identical to the reported perforin cDNA cloned from T cells. Western blot analysis using monoclonal and polyclonal antiperforin peptide antibodies revealed a protein of 65 kD in both human fetal astrocyte and rat natural killer cell lysates (n = 4). Immunostaining followed by FACS(R) and confocal and electron microscopy analysis revealed that perforin was expressed by 40-50% of glial fibrillary acidic protein positive cells present in the fetal brain culture (n = 11). Perforin was not localized to granules in astrocytes but was present throughout the cytoplasm, probably in association with the endoplasmic reticulum. Perforin was not detected in normal adult brain tissue but was present in and around areas of inflammation (white and grey matter) in multiple sclerosis and neurodegenerative brains. Perforin-positive cells were identified as reactive astrocytes. These findings demonstrate that perforin expression is not unique to lymphoid cells and suggest that perforin produced by a subpopulation of astrocytes plays a role in inflammation in the brain.

Long-term effects of cysteamine on cognitive and locomotor behavior in rats: relationship to hippocampal glial pathology and somatostatin levels

Peroxidase-positive astrocytic inclusions, derived from effete, iron-laden mitochondria, accumulate in the rat hippocampus, striatum and other subcortical brain regions as a function of advancing age. The sulfhydryl agent, cysteamine (CSH), accelerates the appearance of this senescent glial phenotype both in primary astrocyte cultures and in the aging subcortical brain in situ. Earlier experiments have shown that short-term administration of CSH results in reversible depletion of brain somatostatin (SS) levels, cognitive deficits and decreases in locomotor activity. In the present study, we tested spatial learning/memory and motor functioning in rats at 4–5 weeks following cessation of chronic (6 week) CSH treatment to determine whether behavioral deficits may be associated with gliopathic changes within the dorsal hippocampus distinct from the behavioral abnormalities accruing to the immediate effects of the drug. CSH-treated rats displayed significantly impaired performance in the Morris water maze 4–5 weeks following termination of prolonged CSH treatment. In contrast, locomotor activity was not affected in this experimental paradigm. CSH-treated animals exhibited significantly higher numbers of peroxidase-positive astrocyte granules as well as total numbers of GFAP-positive astrocytes in the CA1 sector of the dorsal hippocampus relative to saline-treated controls. In the hilus of the dentate gyrus, numbers of both peroxidase-positive glial inclusions and astrocytes were unaffected by CSH exposure. At 5 weeks following cessation of CSH treatment, SS levels in the hippocampus and hypothalamus (but not cerebral cortex) were elevated relative to those of saline-treated controls. Our results indicate that chronic CSH exposure induces senescence-like changes in CA1 astrocytes which are associated with deficits in cognitive, but not locomotor, behavior and elevated levels of hippocampal and hypothalamic SS. Pathological glial-neuronal interactions within the hippocampus and other subcortical brain regions may play an important role in the cognitive decline observed during normal senescence and in aging-related neurodegenerative disorders.

Experimental induction of corpora amylacea-like inclusions in rat astroglia.

Corpora amylacea (CA) are glycoproteinaceous inclusions that accumulate in the human central nervous system during normal ageing, and to an even greater extent in Alzheimer's disease and other neurodegenerative disorders. They are particularly prominent in subpial and subependymal regions, and are most commonly located within astrocytes and their processes. We previously demonstrated that human CA share many tinctorial and histochemical properties in common with Gomori-positive cytoplasmic granules which accumulate in periventricular astrocytes of the ageing vertebrate brain and in rat astroglial cultures exposed to the sulphydryl agent, cysteamine (CSH). In the present study, long-term exposure of neonatal rat astrocyte cultures to CSH resulted in the formation of large spherical, PAS-positive cytoplasmic inclusions which are highly reminiscent of, if not identical to, human CA. As in the case of human CA and Gomori-positive astrocyte granules, the CSH-induced CA-like inclusions exhibit non-enzymatic peroxidase activity and consistent immunolabelling with antibodies directed against the mitochondrial protein, sulphite oxidase. Taken together, our findings suggest that progressive mitochondrial damage and macroautophagy play an important role in the biogenesis of CA (and Gomori-positive granules) in astrocytes of the ageing periventricular brain.

Mitochondrial constituents of corpora amylacea and autofluorescent astrocytic inclusions in senescent human brain

Corpora amylacea (CA) are cytoplasmic inclusions that accumulate in human brain in the course of normal aging, and to an even greater extent, in Alzheimer's disease and other neurodegenerative conditions. In senescent and Alzheimer-diseased human brains, astrocytes in limbic and periventricular regions exhibit red autofluorescent inclusions, homologous to Gomori-positive astrocyte granules previously described in the brains of aging rodents and other vertebrates. We have shown that Gomori inclusions in situ and in culture are derived from autophagocytosed mitochondria exhibiting iron-mediated peroxidase activity. In the human brain, the autofluorescent inclusions share many properties with CA. Both types of inclusion progressively accumulate in periventricular regions with advancing age, are largely astrocytic in origin, and contain various heat shock proteins and ubiquitin. Using histochemistry in conjunction with cofocal microscopy, we demonstrated that both CA and the red autofluorescent granules exhibit non-enzymatic peroxidase activity and an affinity for CAH and PAS. The only major divergent histochemical feature between the Gomori-positive astrocyte granules and CA is the presence of orange-red autofluorescence in the former and the absence of endogenous fluorescence in the latter. On the basis of numerous shared topographic and histochemical features, we hypothesized that CA are largely derived from autofluorescent (Gomori-positive) astrocyte granules which reside in periventricular regions of the senescent CNS. Immunofluorescent labeling and laser scanning confocal microscopy demonstrated consistent colocalization of the mitochondrial proteins, sulfite oxidase, and heat shock protein 60, to both CA and the autofluorescent astroglial inclusions. In addition, both CA and the autofluorescent astrocyte granules exhibit staining for DNA which colocalizes to mitochondrial antigens and therefore likely represents mitochondrial nucleic acid in dual-labeled preparations. These observations suggest that a) Gomori-positive astrocyte granules in human brain are homologous to those described in rodents, b) Gomori-positive granules may be structural precursors of CA in senescent human brain, and c) in the aging human brain, degenerate mitochondria within periventricular astrocytes give rise to autofluorescent cytoplasmic granules and corpora amylacea.

The origin and composition of peroxidase-positive granules in cysteamine-treated astrocytes in culture

Gomori astrocytes, which are prominent in periventricular regions of the brain, contain inclusions that stain with Gomori dyes, and exhibit an orange-red autofluorescence and a non-enzymatic peroxidase activity. Recently, such astrocytes have been induced in dispersed glial cultures by exposure to cysteamine. Using these cells, we have shown that the peroxidase-positive inclusions (Gomori bodies) are multicompartmental, that iron co-localizes with the peroxidase activity, and that the iron is often segregated in one of the compartments of the body. The goal of the present study was to determine the origin and process of formation of these bodies. The results indicate that cysteamine induces aberrations in mitochondrial structure associated with the acquisition of iron and the associated peroxidase activity. Mitochondria thus transformed appear to initiate an autophagic process in which they, and adjacent structures, are sequestered. The presence of acid phosphatase activity in a number of mature Gomori bodies attests to the participation of lysosomal elements in this process. These results indicate, therefore, that the Gomori body is a complex autophagosome in which the iron-containing compartments, putatively responsible for the peroxidase activity, represent undegraded transformed mitochondria.

Stress protein co-localization to autofluorescent astrocytic inclusions in situ and in cysteamine-treated glial cultures

In the aging brain, a unique subpopulation of limbic and perientricular astrocytes accumulates red autofluorescent, peroxidase-positive cytoplasmic inclusions distinct from lipofuscin. Cysteamine (CSH) exposure rapidly induces identical inclusions in cultured, immature astroglia. CSH induces a cellular stress response prior to astrocyte granulation. To determine whether stress proteins are actual consituents of the autofluorescent granules, 12-week-old rat brain sections and CSH-treated astroglial cultures were immunostained with various anti-stress protein antibodies and evaluated by laser scanning confocal microscopy. We observed intense co-localization of heat shock protein (HSP) 27 and ubiquitin (Ub) to the autofluorescent astrocyte granules in situ and in CSH-treated glial cultures. In both preparations, glucose regulated protein (GRP) 94 consistently exhibited partial co-localization to the granule periphery and adjacent cytoplasm. In contrast, HSP72 co-localization to these inclusions was only occasionally seen and the granules appeared entirely devoid of HSP90 and αB-crystallin. Acute exposure of cultured astroglia to CSH induced intense cytoplasmic Ub staining, suggesting that activation of the Ub pathway may be an early event in the biogenesis of these astrocytic granules. Taken together, our results support the notion that the autofluorescent astrocyte inclusions are stress or heat shock granules which progressively accumulated in the aging periventricular brain. Moreover, CSH greatly accelerates the appearance of this senescent astrocyte phenotype in primary culture.

Role of the Cellular Stress Response in the Biogenesis of Cysteamine‐Induced Astrocytic Inclusions in Primary Culture

Cysteamine (CSH; 2-mercaptoethylamine) stimulates the accumulation of peroxidase-positive inclusions in cultured astroglia akin to those observed in the aging periventricular brain. Because CSH induces the synthesis of a stress protein (heme oxygenase) in rat liver, we hypothesized that aspects of the cellular stress response may play a role in the biogenesis of CSH-induced astrocyte granules. In the present study, we performed indirect immunofluorescent staining and immunoblotting for various stress proteins in rat neuroglial cultures. Exposure of astrocyte cultures to CSH enhanced immunostaining for heme oxygenase-1 (HO-1) and heat-shock proteins 27, 72, and 90, but not glucose-regulated protein 94, relative to untreated cultures. CSH-pretreated astrocytes exhibited enhanced tolerance to H2O2 toxicity relative to untreated cells, providing physiological evidence of an antecedent stress response in the former. In addition, exposure for 12 days to H2O2, a known inducer of the stress response, elicited astrocyte granulation similar to that observed with CSH. Chronic induction of HO-1 and other stress proteins may participate in the biogenesis of metalloporphyrin-rich inclusions in CSH-treated astroglial cultures and in astrocytes of the aging periventricular brain.

Colocalization of Organelle-Specific Proteins to Autofluorescent Astrocyte Granules by Laser Scanning Confocal Microscopy

Astrocytes in aging periventricular brain regions and in cysteamine-treated neonatal brain cell cultures contain cytoplasmic inclusions that exhibit an affinity for Gomori stains, orange-red autofluorescence, and nonenzymatic peroxidase activity. In order to delineate the cellular constituents participating in the biogenesis of these astrocytic inclusions, colocalization of FITC-immunolabeled organelles to the red autofluorescent granules was analyzed using a laser scanning confocal imaging system. Areas of true colocalization exhibited yellow fluorescence which persisted in Z-axis image reconstructions. We observed no colocalization of catalase- and PMP70-positive peroxisomes or the Golgi apparatus to the autofluorescent inclusions. The rough endoplasmic reticulum exhibited infrequent dot-like regions of colocalization consistent with previous electron microscopic observations. A minority of early endosomes colocalized to the autofluorescent inclusions in perinuclear cytoplasm. There was extensive colocalization of lgp120-labeled lysosomes to larger autofluorescent granules in close proximity to the nucleus whereas smaller autofluorescent granules often remained unlabeled. A macroautophagic process involving lysosomes and to lesser extent, early endosomes and the rough endoplasmic reticulum, may participate in the biogenesis of autofluorescent astrocytic inclusions in cysteaminetreated glial cultures and in the aging periventricular brain.

Isolation of pseudoperoxidase-positive astrocyte granules from intact rat brain and cysteamine-treated neuroglial cultures

A subpopulation of astrocytes in periventricular regions of aging brain and in cysteamine (CSH)-treated glial cultures contain autofluorescent cytoplasmic granules that exhibit an affinity for Gomori's chrome alum hematoxylin (CAH), and non-enzymatic peroxidase activity. Although shown to be histochemically distinct from lipofuscin, the lack of pure preparations of these glial inclusions has hindered the elucidation of their precise chemical constituents. Using sucrose gradient fractionation and density centrifugation on percoll, we obtained enriched preparations of astrocyte cytoplasmic granules from intact rat brain and CSH-treated astrocyte cultures. The presence and relative purity of these inclusions were confirmed by laser scanning confocal microscopy for red autofluorescent granules, diaminobenzidine histochemistry for non-enzymatic peroxidase activity and chrome alum hematoxylin (Gomori) staining. In the enriched fractions, the smaller granules (0.5-4.0 microns) were spherical and weakly autofluorescent, whereas larger inclusions (5.0-10.0 microns) tended to be intensely autofluorescent and pleomorphic. As in situ, the purified material was argyrophilic and did not stain for lipids. Isolation of these astrocytic inclusions should permit a more thorough characterization of their biochemical contents.

Gomori‐positive astrocytes: Biological properties and implications for neurologic and neuroendocrine disorders

Granule laden astrocytes exhibiting an affinity for chrome alum hematoxylin and aldehyde fuchsin (Gomori stains) have been described in the periventricular brain of all terrestrial vertebrate species examined to date including humans. The astrocytic inclusions are rich in sulfhydryl groups, emit an orange-red autofluorescence, and stain intensely with diaminobenzidine, a marker of endogenous peroxidase activity. The distinct autofluorescence pattern and the absence of neutral lipid, acid phosphatase, and beta-glucuronidase activity exclude lipofuscin or lysosomes as components of these astrocytic granules. The emission of orange-red autofluorescence and the nonenzymatic nature of the peroxidase activity implicate the presence of porphyrins and metalloporphyrins such as heme as major constituents of these cytoplasmic gliosomes. The role of Gomori-positive astrocytes under normal and pathologic conditions is incompletely understood. In vivo, numbers of astrocytic granules increase as a function of advancing age, in response to chronic estrogen stimulation, and following X-irradiation. In vitro, these cells accumulate with increasing time in culture and following exposure to the sulfhydryl agent, cysteamine. Gomori-positive astrocytes may supply heme to neurons for the synthesis of cytochromes, catalases, and other heme enzymes. They may play a role in photostimulation of sexual cyclicity, the promotion of neuritic development, the degradation of toxic lipoperoxides, and the metabolism of various neurotransmitters. Conversely, these cells may contribute to the pathogenesis of several neurologic and neuroendocrine disorders. Examples of the latter include a) augmentation of goldthioglucose neurotoxicity, b) induction of hypothalamic anovulation and reproductive failure, c) exacerbation of porphyric encephalopathy, and d) potentiation of parkinsonism and other free radical-related neurodegenerations.

Effects of progestins on the estradiol-related accumulation of astrocytic granules in the hypothalamic arcuate nucleus

Adult female rats treated with a single 2 mg injection of estradiol valerate (EV) develop anovulatory sterility (persistent estrus) and neuropathologic changes in the hypothalamic arcuate nucleus. A prominent feature of the latter is the accumulation of peroxidase-positive cytoplasmic inclusions in arcuate astrocytes which results from aberrant patterns of chronic, ovarian E 2 secretion in this model. In the present study, we tested an hypothesis that progesterone may antagonize the pathologic effects of E 2 within the medial basal hypothalamus analogous to effects in peripheral steroid target tissues. In intact EV-treated rats, we observed complete suppression of the astrocytic reaction by subsequent injections of medroxyprogesterone acetate (MPA). However, chronic progesterone exposure did not significantly antagonize this estrogenic affect in ovariectomized rats implanted subcutaneously with steroid-releasing Silastic capsules. Taken together, these results suggest that progestins may block the development of an E 2-related arcuate lesion by suppressing the pituitary-ovarian axis rather than by directly antagonizing the dystrophic effects of E 2 at the hypothalamic level.

Ovary-Dependent Degeneration in the Hypothalamic Arcuate Nucleus*

The effects of estradiol valerate and constant light exposure on the histological appearance of the arcuate nucleus were assessed in female rats. Both of these treatments caused significant increases in the numbers of reactive microglial cells and astrocytic granules in the nucleus. Ovariectomy before either treatment prevented the glial reaction, indicating that the experimental manipulations triggered the secretion of an ovarian product which appears to be selectively toxic to the arcuate nucleus. The fact that monthly injections of estradiol valerate in male rats produced the same profile of degeneration in the arcuate nucleus suggests that the neuropathological agent may itself be estradiol. Ovariectomy also significantly reduced arcuate microglial reactivity associated with normal aging, which suggests that cyclic surges of endogenous estradiol may be capable of gradually producing an arcuate lesion. This phenomenon may accout for the hypothalamic reproductive failure associated with normal aging in the rat.