共查询到20条相似文献,搜索用时 0 毫秒
1.
Thyroidal stimulation of tubulin and actin in primary cultures of neuronal and glial cells of rat brain 总被引:1,自引:0,他引:1
The influence of triiodothyronine (T3) on the level of tubulin and other proteins in primary cultures of neuronal (N) and glial (G) cells from rat brain has been investigated. Quantitation of tubulin by [3H]colchicine binding assay revealed that when cells from 1 day rat brain were cultured for 18 hr with physiological doses (0.5-5 nM) of T3, the hormone elicited 35-40% increase in the soluble (30,000 g supernatant) tubulin content of G cells only. This stimulation was age-dependent and occurred neonatally at a time corresponding to the onset of synaptogenesis. In mouse and chick brain also, [3H]colchicine binding assay showed a similar selective stimulation of the soluble tubulin content of G cells by T3 with virtually no effect on N cells. However, SDS-polyacrylamide gel electrophoresis of the total proteins in the 30,000 g supernatants from N and C cells of rat brain, labeled for 18 hr with [14C]leucine in the presence of T3, revealed that T3 elicited 2-3-fold enhancement of radiolabeled tubulin in the N cells which is relatively greater than the 1.5-fold increase seen in the G cells. Analysis of the autoradiograms of these labeled proteins also revealed that in addition to tubulin, T3 stimulated the accumulation of radiolabeled actin by 1.5- and 2-fold in N cells and G cells respectively. Similar electrophoretic analysis of the solubilized labeled proteins in the 30,000 g pellets from N and G cells indicated that the failure to detect the stimulation of tubulin in the 30,000 g supernatants from N cells by [3H]colchicine binding assay could be at least partly due to rapid translocation of the dimeric soluble tubulin into insoluble membrane fractions or due to presence of higher oligomeric forms of tubulin which are insensitive to [3H]colchicine binding assay. 相似文献
2.
3.
Exposure of organ cultures of newborn rat brains to tri-iodothyronine (T3) followed by cell fractionation as well as direct exposure of prefractionated neuronal (N) and glial (G) cells to the hormone results in an almost selective induction of tubulin in the glial cells. This is established from two independent assays of tubulin, viz. colchicine binding and vinblastin precipitation. In the newborn rat brain, the tubulin content of the G cells is almost 3-fold higher than that of the N cells. Treatment with T3 elicits 40-50% stimulation of tubulin in the G cells within 2 hr without any significant increase in the N cells. Brains from 8- or 50-day-old rats are irresponsive to induction to tubulin by T3. The rate of incorporation of [3H]leucine into total protein is very similar in both N and G cells of newborn rat brain but that into tubulin of G cells is about 3-fold higher than that of N cells. T3 promotes this incorporation by over 30% in the G cells with only a marginal 5% increase in the N cells. The overall results suggest that the glial cells represent the target cells for the T3-induced synthesis of tubulin, the major structural protein of the developing brain. 相似文献
4.
5.
Cognitive aspects of the acute stress response are partly mediated through activation of the locus coeruleus (LC)-norepinephrine (NE) system via corticotropin-releasing factor (CRF). Apart from mediating the acute responses to stress, CRF can mediate the long-term impact of stress on the brain through its potent modulation of neuronal morphology. Importantly, the cellular pathways engaged by stress in general, and CRF in particular, in remodeling neuronal structure are poorly understood. Here, we demonstrate that apart from its well-established acute effects on LC neuronal activity, CRF also stimulates growth and arborization of LC neuronal processes. By contrast, urocortin 2 (UCN 2), a related peptide, inhibits outgrowth of such processes. These opposing effects are transduced by a common receptor (CRF(1)) but distinct intracellular signaling pathways. The structural effects of CRF required protein kinase A and mitogen-activated protein kinase, as well as Rac1, a member of the Rho family of GTPases that regulates the actin and microtubule cytoskeleton. By contrast, the effects of UCN II were mediated by the protein kinase C and RhoA pathways. This is the first study to link stress-related substrates to molecular mediators of actin cytoskeletal remodeling in the LC. We propose a model of dynamic LC neuronal plasticity that is reciprocally controlled by CRF and UCN II, eventually determining actin rearrangement by Rho-specific pathways. By regulating the extension of processes into pericoerulear regions where limbic afferents terminate, these peptides may determine the degree to which the LC-NE system is influenced by limbic structures that mediate emotional expression. 相似文献
6.
Chauvet N Prieto M Fabre C Noren NK Privat A 《Molecular and cellular neurosciences》2003,22(4):467-486
p120 catenin (p120ctn) is implicated in the regulation of cadherin-mediated adhesion and actin cytoskeleton remodeling. The interaction of cytoplasmic p120ctn with the guanine exchange factor Vav2 is one of the signaling pathways implicated in cytoskeleton dynamics. We show here that p120ctn is regulated during rat brain development and is distributed at the membrane and within the cytoplasm where it associates with N-cadherin and Vav2, respectively. p120ctn shifts progressively from an axonal expression to a punctuate staining localized to a subset of synapses. In cultured hippocampal neurons, p120ctn redistributes from growth cones to synapses, where it partly colocalizes with N-cadherin or Vav2 and filamentous actin. In the adult forebrain, we show that p120ctn and Vav2 are highly expressed by neuroblasts migrating from the lateral subventricular zone to the olfactory bulb. The dynamic expression pattern of p120ctn and the biochemical evidences of its association with N-cadherin and Vav2 strongly suggest that p120ctn plays a major role in neuronal migration, neurite outgrowth and synapse formation, and plasticity. 相似文献
7.
8.
Kate E. Keller 《中国神经再生研究》2020,15(11):2031
正Glaucoma and the actin cytoskeleton: Glaucoma is an optic neuropathy, with pathophysiological changes affecting anterior and posterior tissues of the eye. The trabecular meshwork(TM) in the anterior segment regulates intraocular pressure(IOP), while photoreceptors in the posterior retina convert light into 相似文献
9.
A monoclonal antibody, G8, was isolated which recognizes a form of tubulin (G8-tubulin) with a novel distribution in the rat brain. Immunoblots of rat brain homogenates and immunohistochemical staining of rat brain sections of various ages with G8 revealed a restricted and developmentally regulated distribution of the G8-tubulin. G8 staining was primarily found in granule cell dendrites in the dentate gyrus, in pyramidal cell apical dendrites in the hippocampus, and in Purkinje cell dendrites in the cerebellum. This pattern was much more selective than that observed with three other anti-tubulin antibodies. Relative abundance of G8-tubulin in the brain increases with age between postnatal day 1 (P1) and adulthood. The results suggest that G8 is specific for a novel tubulin form which shows a characteristic distribution in the rat brain. This distribution may indicate that the G8-tubulin possesses functional specificity. 相似文献
10.
The distribution of mGlu1 alpha receptor and tubulin was immunocytochemically examined in the rat cerebellar cortex and primary rat cortical neurons at both immunofluorescence and electron microscopic level. In cryosections from rat cerebellar cortex mGlu1 alpha receptor immunoreactivity was expressed in cell bodies and dendrites of Purkinje and basket cells of the cerebellar molecular layer. Tubulin immunoreactivity was concentrated in the dendritic tree of the cerebellar molecular layer, as well as in the granule cell layer. In primary rat cortical neurons, both proteins colocalized throughout the proximal and distal dendrites of these cells. At the electron microscopic level, the receptor was present in dendritic shafts and dendritic spines of Purkinje cells at perisynaptic sites of asymmetrical synapses. Immunoreactivity corresponding to tubulin was associated with the plasma membrane of dendritic shafts of Purkinje cells, as well as throughout its cytoplasm as part of the cytoskeletal components. Interestingly, double labeling for both proteins reveals an association of tubulin with mGlu1 alpha receptor at the plasma membrane level of dendritic shafts of Purkinje cells. This suggests that tubulin interacts with mGlu1 alpha receptor and may be involved in the anchoring of the receptor to the plasma membrane. 相似文献
11.
Representing the most common cause of dementia, Alzheimer's disease (AD) has dramatically impacted the neurological and economic health of our society. AD is a debilitating neurodegenerative disease that produces marked cognitive decline. Much evidence has accumulated over the past decade to suggest soluble oligomers of beta-amyloid (Aβ) have a critical role in mediating AD pathology early in the disease process by perturbing synaptic efficacy. Here we critically review recent research that implicates synapses as key sites of early pathogenesis in AD. Most excitatory synapses in the brain rely on dendritic spines as the sites for excitatory neurotransmission. The structure and function of dendritic spines are dynamically regulated by cellular pathways acting on the actin cytoskeleton. Numerous studies analyzing human postmortem tissue, animal models and cellular paradigms indicate that AD pathology has a deleterious effect on the pathways governing actin cytoskeleton stability. Based on the available evidence, we propose the idea that a contributing factor to synaptic pathology in early AD is an Aβ oligomer-initiated collapse of a "synaptic safety net" in spines, leading to dendritic spine degeneration and synaptic dysfunction. Spine stabilizing pathways may thus represent efficacious therapeutic targets for combating AD pathology. 相似文献
12.
Ram P. Gupta Mohamed B. Abou-Donia 《Molecular and chemical neuropathology / sponsored by the International Society for Neurochemistry and the World Federation of Neurology and research groups on neurochemistry and cerebrospinal fluid》1997,30(3):223-237
Acrylamide and carbon disulfide produce central-peripheral distal axonopathy in experimental animals and humans. The main feature of this disease is the focal swellings containing neurofilaments in distal axons, followed by nerve degeneration beyond these swellings. We studied the possible role of tubulin assembly kinetics in this disease. The rats were either administered acrylamide (50 mg/kg, ip, saline) or exposed to carbon disulfide (700 ppm, 9h) via inhalation for 12 and 15 d, respectively. Tubulin, purified from both acrylamide-(10.37±0.3 vs 11.3±0.15) and carbon disulfide-treated (9.72±0.5 vs 11.18±0.25) rat brains showed increase inV max (OD/min × 103) of its polymerization. However, only acrylamide treatment showed a decrease in time toV max, when brain supernatant was used for tubulin polymerization. In vitro addition of acrylamide (0.1–1 mM) to bovine brain tubulin also showed a decrease in time toV max (16–21%) of its polymerization. Carbon disulfide treatment of rats, on the other hand, showed a decrease in MAP-2 and an increase in a 120-kDa peptide concentration. The latter showed immunoreactivity with anti-MAP-2. The increase in the rate of tubulin polymerization by acrylamide and carbon disulfide treatment may alter the rate of transport of axonal consituents, including neurofilament, and contribute toward their accumulation in the focal swellings observed in this neuropathy. 相似文献
13.
The neurofibromatosis 2 (NF2) suppressor gene encodes a protein termed merlin (or schwannomin) with sequence similarity to a family of proteins that link the actin cytoskeleton to cell surface glycoproteins. Members of this ERM family of proteins include ezrin, radixin, and moesin. These proteins contain a carboxyl (C-) terminus actin binding site. In contrast to the ERM proteins, merlin lacks the conventional C-terminal actin binding site, but still localizes to the ruffling edge of plasma membranes. In this study, we investigate the ability of merlin to interact with actin through a nonconventional actin binding domain. We demonstrate for the first time that merlin can associate with polymerized actin in vitro by virtue of an amino (N-) terminal actin binding domain including residues 178–367. Merlin actin binding is not affected by several naturally-occurring NF2 patient mutations or alternatively spliced isoforms. These results suggest that merlin, like other ERM proteins, can directly interact with the actin cytoskeleton. In addition, merlin associates with polymerized microtubules in vitro using a novel microtubule binding region in the N-terminal region of merlin that is masked in the full-length merlin molecule, such that wild-type functional merlin in the “closed” conformation fails to bind polymerized microtubules. These microtubule association results confirm the notion that merlin exists in “open” and “closed” conformations relevant to its function as a negative growth regulator. J. Neurosci. Res. 51:403–415, 1998. © 1998 Wiley-Liss, Inc. 相似文献
14.
Kuipers HF Rappert AA Mommaas AM van Haastert ES van der Valk P Boddeke HW Biber KP van den Elsen PJ 《Glia》2006,53(2):115-123
Statin treatment is proposed to be a new potential therapy for multiple sclerosis (MS), an inflammatory demyelinating disease of the central nervous system. The effects of statin treatment on brain cells, however, are hardly understood. We therefore evaluated the effects of simvastatin treatment on the migratory capacity of brain microglial cells, key elements in the pathogenesis of MS. It is shown that exposure of human and murine microglial cells to simvastatin reduced cell surface expression of the chemokine receptors CCR5 and CXCR3. In addition, simvastatin treatment specifically abolished chemokine-induced microglial cell motility, altered actin cytoskeleton distribution, and led to changes in intracellular vesicles. These data clearly show that simvastatin inhibits several immunological properties of microglia, which may provide a rationale for statin treatment in MS. 相似文献
15.
Low frequency stimulation modifies receptor binding in rat brain 总被引:4,自引:0,他引:4
Experiments were designed to reproduce the antiepileptic effects of low frequency stimulation (LFS) during the amygdala kindling process and to examine LFS-induced changes in receptor binding levels of different neurotransmitters in normal brain. Male Wistar rats were stereotactically implanted in the right amygdala with a bipolar electrode. Rats (n = 14) received twice daily LFS (15 min train of 1Hz, 0.1 ms at an intensity of 100 to 400 microA) immediately after amygdala kindling stimulation (1s train of 60 Hz biphasic square waves, each 1 ms at amplitude of 200-500 microA) during 20 days. The LFS suppressed epileptogenesis (full attainment of stage V kindling) but not the presence of partial seizures (lower stages of kindling) in 85.7% of the rats. Thereafter, normal rats (n = 7) received amygdala LFS twice daily for 40 trials. Animals were sacrificed 24 h after last stimulation and their brain used for labeling mu opioid, benzodiazepine (BZD), alpha(1)-adrenergic, and adenylyl cyclase binding. Autoradiography experiments revealed increased BZD receptor binding in basolateral amygdala (20.5%) and thalamus (29.3%) ipsilateral to the place of stimulation and in contralateral temporal cortex (18%) as well as decreased values in ipsilateral frontal cortex (24.2%). Concerning mu receptors, LFS decreased binding values in ipsilateral sensorimotor (7.2%) and temporal (5.6%) cortices, dentate gyrus (5.8% ipsi and 6.8% contralateral, respectively), and contralateral CA1 area of dorsal hippocampus (5.5%). LFS did not modify alpha(1) receptor and adenylyl cyclase binding values. These findings suggest that the antiepileptic effects of LFS may involve activation of GABA-BZD and endogenous opioid systems. 相似文献
16.
Analgesia from rostral brain stem stimulation in the rat 总被引:1,自引:0,他引:1
Rats implanted with bipolar stimulating electrodes in the rostral medial brain stem were tested for brain stimulation-produced analgesia using tail-flick, pinch and hot-plate tests. Potent analgesia across all three tests was obtained from stimulation of sites in the gray matter surrounding the aqueduct and the caudal portion of the third ventricle, the posterior hypothalamus, the midline area of the caudal thalamus and the pretectal region of the meso-diencephalic junction. The analgesia obtained from these sites was comparable to that produced by stimulation of the previously studied caudal periaqueductal gray matter: it outlasted the period of brain stimulation, was not due to a generalized motor debilitation of the animal, and was not correlated with changes in electrographic activity. Stimulation of sites in the caudal thalamus and pretectal area yielded analgesia without stimulation-induced aversive reactions, confirming the potential of these sites for use in the relief of clinical pain in man. 相似文献
17.
Presenilin I (PSI) has been shown to interact with microfilament-associated proteins of the filamin family. Here, we investigated a possible association of PSI with the cytoskeleton. Immunoblotting of detergent-insoluble fractions of rat brain homogenate revealed enrichment of neuron-specific 36 and 14 kDa proteolytic fragments of PSI, whereas 30 and 20 kDa fragments were found in the detergent-soluble fraction. Specific severing of microfilaments with gelsolin in the detergent-insoluble pellet and subsequent centrifugation led to the detection of both actin and PSI fragments in the supernatant. In addition, in vitro translated PSI cosedimented with actin filaments. Our findings provide biochemical evidence for the association of PSI fragments with actin filaments. 相似文献
18.
Christensen RN Ha BK Sun F Bresnahan JC Beattie MS 《Journal of neuroscience research》2006,84(1):170-181
Microglia are key mediators of the immune response in the central nervous system (CNS). They are closely related to macrophages and undergo dramatic morphological and functional changes after CNS trauma or excitotoxic lesions. Microglia can be directly stimulated by excitatory neurotransmitters and are known to express many neurotransmitter receptors. The role of these receptors, however, is not clear. This study describes the microglial response to the glutamate receptor agonist kainate (KA) and shows via immunochemistry that the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-type glutamate receptor subunit GluR1 is present on cultured microglia. In the presence of 100 microM or 1 mM KA, cultured microglia underwent dramatic morphological and cytoskeletal changes as observed by time-lapse photography and quantitative confocal analysis of phalloidin labeling. KA-stimulated microglia showed condensation of cytoplasmic actin filaments, rapid de- and repolymerization, and cytoplasmic redistribution of condensed actin bundles. Rearrangement of actin filaments-thought to be involved in locomotion and phagocytosis and to indicate an increased level of activation (for reviews see Greenberg [ 1995] Trends Cell Biol. 5:93-99; Imai and Kohsaka [ 2002] Glia 40:164-174)-was significantly increased in treated vs. control cultures. Morphological plasticity and membrane ruffling were also seen. These findings suggest direct microglial excitation via glutamate receptor pathways. Thus, neurotransmitter release after brain or spinal cord injury might directly modulate the inflammatory response. 相似文献
19.
20.
The slow component (SC) of axonal transport conveys structural proteins, regulatory proteins, and glycolytic enzymes toward the axon tip at 1–6 mm/day. Following axon interruption (axotomy), the rate of outgrowth corresponds to the rate of SCb—the fastest subcomponent of SC. Both axonal outgrowth and SCb accelerate 20–25% after axotomy. Tubulin and actin are the major proteins being carried by SCb. To further characterize the acceleration of SCb, we measured the equilibrium between subunits and polymers for both actin and tubulin. We radiolabeled newly synthesized proteins in rat motor neurons by microinjecting [35S]methionine into the spinal cord 7 days after crushing the sciatic nerve (85 mm from the spinal cord). Nerves were removed 7 days later for homogenization in polymer-stabilizing buffer (PSB) and centrifugation, followed by SDS-PAGE of supernatants (S) and pellets (P). We removed β-tubulin, actin, and the medium-weight neurofilament protein (NF-M) from each gel by using the fluorogram as a template. After solubilizing gel segments for liquid scintillation spectrometry, we expressed counts as a polymerization ratio: P/[S + P]. In the nerve segments that contained radiolabeled SCb proteins, located 24–36 mm from the spinal cord, axotomy increased the polymerization ratio of SCb actin from 0.23 to 0.36 (P < 0.05) but had no effect on SCb β-tubulin. In a separate experiment, we added 12 μM taxol to PSB to stabilize newly assembled microtubules. Adding taxol did not alter the polymerization ratio for SCb β-tubulin in sham-axotomized nerves but did increase the ratio in axotomized nerves, from 0.44 to 0.63 (P < 0.05); polymerization ratios for SCb actin were unaffected. We conclude that the assembly of microfilaments and microtubules increases to provide cytoskeletal elements for axon sprouts. The resulting loss of actin and tubulin subunits may play a role in the acceleration of SCb. © 1996 Wiley-Liss, Inc. 相似文献