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Biomedical subjects

U Otten

Publications and source records attributed to U Otten.

At least 73 records · Page 4Linked to original sources
PubMed1993

Glucocorticoid modulation of neurotrophin expression in immortalized mouse hippocampal neurons.

To see whether glucocorticoid hormones can influence the regulation of neurotrophin expression in hippocampal neurons, we have used reverse-transcription followed by polymerase chain reaction to investigate changes in the mRNA levels of nerve growth factor (NGF) and neurotrophin-3 (NT-3) in immortalized hippocampal neurons after dexamethasone application. Our results show that NGF mRNA levels rise in both embryonic and postnatal neurons, but with different time courses, while NT-3 levels rise in the embryonic but not in the postnatally derived cell line. Modulation of NT expression by glucocorticoids may therefore be developmentally regulated.

Animals↗
PubMed1993

Effects of perinatal hypo- and hyperthyroidism on the levels of nerve growth factor and its low-affinity receptor in cerebellum.

Deficits or excesses of thyroid hormones during critical periods of mammalian cerebellar development can lead to profound biochemical and morphological abnormalities in this system. The goal of this study was to investigate the effects of perinatal hypo- and hyperthyroidism on the ontogeny of nerve growth factor (NGF) and its low-affinity receptor (p75NGFR) in the rat cerebellum. The concentration of NGF and of p75NGFR immunoreactivity (IR) were measured, several days after birth, in cerebella of rats which had received propylthiouracil (PTU) or thyroxine. NGF concentration was markedly enhanced only on postnatal day 2 (P2) in hyperthyroid rats, whereas in hypothyroid (PTU-treated) rats NGF values were similar to age-matched controls. These observations suggest that thyroid hormone affects NGF synthesis during early periods of cerebellar development. In Purkinje cells of control animals, p75NGFR IR peaked at P10. In hypothyroid rats, the expression of p75NGFR was retarded, peaking at P15, whereas in hyperthyroid rats it was advanced, peaking at P8. The increased p75NGFR IR found in Purkinje cell bodies and the delayed disappearance of p75NGFR IR from the external granular layer of hypothyroid rats suggest different roles for thyroid hormone in the developing cerebellum. We conclude that p75NGFR and NGF are independently regulated by thyroid hormone during critical periods of cerebellar development. The effect of thyroid hormone deficiency on p75NGFR content in Purkinje cells may involve complex mechanisms such as impaired efficiency of axonal transport.

Animals↗
PubMed1993

Differential expression of interleukin-6 (IL-6) and interleukin-6 receptor (IL-6R) mRNAs in rat hypothalamus.

Using the sensitive technique of reverse transcription followed by polymerase chain reaction (RT-PCR), the expression of interleukin-6 (IL-6) and its receptor (IL-6R) in rat hypothalamus was investigated. Our results show a differential expression of the mRNAs for IL-6 and IL-6R during postnatal development: whereas IL-6 mRNA slightly declines with increasing age, IL-6R mRNA levels markedly increase (4-fold). The time course of these changes correlates with the maturation of the hypothalamo-pituitary-adrenocortical axis, indicating an involvement of the IL-6 system in neuroendocrine regulation. Expression of both genes in other brain areas suggest additional functions in the central nervous system (CNS).

Aging↗
PubMed1993

Spreading depression induces c-fos-like immunoreactivity and NGF mRNA in the rat cerebral cortex.

Application of potassium chloride (KCl) to the brain surface elicits spreading depression which leads to a marked induction of the proto-oncogene c-fos in the treated cerebral cortex at the earliest time examined (90 min). High levels of c-fos immunoreactivity are observed up to 6 h after KCl treatment. The areas affected include the cingulate, entorhinal and frontoparietal cortex throughout the treated hemisphere. The c-fos expression preceded an increase in both NGFmRNA and NGF-like protein(s). A maximal increase in c-fos was detected within 3 h, whereas NGFmRNA peaked at 12 h and NGF-like protein(s) reached their maximum level 24 h after KCl application. The most prominent increase in NGFmRNA was measured in the entorhinal cortex (50-fold), but other cortical areas also showed a moderate increase of 2-3-fold. In conclusion, our results provide evidence that increases in c-fos and NGF expression are early adaptive responses following brain injury.

Animals↗
PubMed1993

Nerve growth factor (NGF) regulates tachykinin gene expression and biosynthesis in rat sensory neurons during early postnatal development.

The regulatory effects of nerve growth factor (NGF) on tachykinin biosynthesis in rat primary sensory neurons during the period of postnatal development were examined under in vivo and in vitro conditions. Administration of NGF to neonatal rats led to a significant increase in protein levels of substance P (SP) and neurokinin A (NKA) in trigeminal and dorsal root ganglia (DRG). In addition, Northern blot analysis revealed that preprotachykinin mRNA was upregulated in sensory ganglia of neonatal animals after treatment with NGF. Using a well-defined in vitro system for neonatal rat DRG and trigeminal ganglia neurons, we found that addition of NGF induced SP and NKA protein levels in a dose-dependent manner. Furthermore, preprotachykinin mRNA was markedly increased in cultured DRG and trigeminal ganglia neurons in the presence of NGF. Thus, our results clearly demonstrate that NGF regulates tachykinin gene expression and biosynthesis both in vivo and in vitro during the developmental period of rat sensory neurons.

Animals↗
PubMed1993

Increased levels of NGF in sera of systemic lupus erythematosus patients.

Using a specific enzyme-linked immunosorbent assay (ELISA) for human nerve growth factor (NGF), serum levels in patients with systemic lupus erythematosus (SLE) were measured. We found a consistent increase in NGF levels in SLE patients compared with controls. A good correlation exists between serum NGF level and severity of clinical manifestation. We hypothesize that NGF might play a role in the pathogenesis of autoimmune disorders such as SLE.

Adolescent↗
PubMed1992

Multiple sclerosis patients express increased levels of beta-nerve growth factor in cerebrospinal fluid.

We describe the measurement of beta-nerve growth factor (NGF) content in cerebrospinal fluid (CSF) from multiple sclerosis (MS) patients compared with CSF from age-matched normal subjects using a specific sandwich immunoassay (ELISA). During acute attacks patients exhibit a significant increase of NGF content compared to controls. In contrast during remission the mean NGF levels in CSF markedly decrease. These results strongly indicate that increased NGF production in CSF is a characteristic feature of the MS inflammatory response.

Adolescent↗
PubMed1992

Expression of neurotrophins and the low-affinity NGF receptor in septal and hippocampal reaggregate cultures: local physiologic effects of NGF synthesized in the septal region.

Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) are members of a family of trophic factors designated the neurotrophins, each of which can bind to the low-affinity NGF receptor (LNGFR). To investigate the mechanisms that regulate the expression of the neurotrophins and the LNGFR in the developing brain, we grew cells from the embryonic mouse septum and hippocampus in reaggregating cell culture and compared neurotrophin and LNGFR expression in developing reaggregates with that seen in the developing septum and hippocampus in situ. NGF, BDNF, NT-3 and LNGFR were each expressed in septal and hippocampal reaggregates as well as the native septum and hippocampus. Additionally, the temporal expression profiles observed in reaggregates were generally similar to those seen in the respective brain regions in situ. In order to determine whether NGF can modulate neurotrophin or LNGFR expression, reaggregates were cultured in the continual presence of either exogenous NGF or anti-NGF antibodies. NGF-treated septal cultures expressed twice the level of LNGFR mRNA as was seen in untreated septal cultures; on the other hand, septal cultures grown in the presence of anti-NGF antibodies, to neutralize endogenously synthesized NGF, displayed a 3-fold decrease in LNGFR mRNA expression compared to untreated cultures. No effects of NGF or anti-NGF were observed on LNGFR expression in hippocampal reaggregates, or on neurotrophin mRNA expression in either reaggregate type. These results suggest that regulatory mechanisms intrinsic to the septal and hippocampal regions control neurotrophin and LNGFR expression. NGF is likely to be one of these regulatory cues since it acts locally in septal reaggregates to control the developmental expression of LNGFR mRNA. The possible roles of locally synthesized NGF and other neurotrophins in the development of septal neurons are discussed.

Aging↗
PubMed1991

Biosynthesis and release of tachykinins from rat sensory neurons in culture.

A long-term culture system of dissociated rat dorsal root (DRG) and trigeminal ganglion cells with high cell density has been developed. Two to 3 weeks after plating, the cultures consist of a nearly pure population of sensory neurons, which can be kept for more than 2 months in culture. Cultured neurons synthesize and release the tachykinins substance P (SP) and substance K (SK, neurokinin A) with a time course similar to that observed in vivo. High-pressure liquid chromatography (HPLC) analysis of peptides extracted from neuronal cultures and synthetic tachykinins revealed identical retention characteristics. Northern blot analysis of mRNAs from cultured cells with a specific tachykinin-probe demonstrated that the preprotachykinin-gene is expressed in preparations of both DRG and trigeminal ganglia cells. Depolarizing stimuli such as high potassium (47 mM) evoked a peptide release from cultured neurons in a strictly Ca(++)-dependent manner. Capsaicin, a compound known to stimulate nociceptive sensory neurons, dose-dependently released tachykinins in concentrations as low as 10(-9) M. Only total absence of Ca++ ions from the incubation medium abolished the capsaicin-induced peptide release. Nifedipine, a blocker of voltage-dependent L-type Ca++ channels, completely blocked the potassium-induced release of SP but did not reduce the capsaicin-evoked release. Mediator substances of pain and inflammation, such as bradykinin, serotonin, and histamine, triggered the release of tachykinins from sensory neurons in vitro. These results clearly demonstrate that the neurons characterized express properties similar to those of sensory neurons in vivo and provide model systems for detailed studies of the biosynthesis and release of neuropeptides as well as the participation of sensory neurons in pain and inflammatory reactions.

Animals↗
PubMed1991

Interleukin-1 beta and tumor necrosis factor-alpha synergistically stimulate nerve growth factor synthesis in rat mesangial cells.

Recent evidence indicates that cytokines are potent inducers of nerve growth factor (NGF) expression both in peripheral tissues and the central nervous system and that NGF, in addition to its neurotrophic action, also acts as an immunoregulatory agent. It was of interest to investigate whether inflammatory cytokines affect NGF production in renal mesangial cells, which play a crucial role in the modulation of the local immune function in the glomerulus. Our results show that the simultaneous addition of interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) elicited a marked (13-fold) increase of NGF protein released by cultured rat glomerular mesangial cells within 24 h, whereas IL-1 alpha in combination with TNF-alpha, as well as the cytokines alone, did not promote the synthesis of NGF. The synergistic effect was dose dependent (maximal at 1 nM) and due to enhanced gene expression, since the cytokine treatment caused a fivefold increase in NGF mRNA after 8 h. Stimulation of NGF synthesis was abolished by mepacrine and dexamethasone, indicating that phospholipase A2 may be involved in NGF regulation. Moreover, pretreatment of the cells with the lipoxygenase inhibitor nordihydroguaiaretic acid (NDGA) abolished induction of NGF by cytokines; in contrast, the specific cyclooxygenase inhibitors indomethacin and diclofenac failed to modify NGF production. These data suggest that a lipoxygenase metabolite produced in response to IL-1 beta and TNF-alpha acts as a mediator in NGF gene expression. In conclusion, these findings support a model in which a cytokine cascade including NGF may play an important role in the pathophysiology of inflammatory renal diseases.

Alkaloids↗
PubMed1990

Interleukin-1 beta and tumor necrosis factor-alpha synergistically stimulate nerve growth factor (NGF) release from cultured rat astrocytes.

Nerve growth factor (NGF) may mediate responses to brain injury. To examine regulation of NGF gene expression with respect to neural trauma we examined the effects of interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) on NGF production in cultures of rat astroglial cells. Purified neocortical astrocytes in serum-free medium were treated with IL-1 beta, TNF-alpha or both. Whereas IL-1 beta and TNF-alpha alone elicited only small effects, simultaneous addition elicited within 48 h a large (3- to 6-fold) increase in NGF content in culture supernatants. Our data are consistent with a role for cytokines in NGF synthesis and release in the injured central nervous system (CNS).

Animals↗
PubMed1990

Measurement of human nerve growth factor binding sites in brain and in peripheral tissues by a specific immunoprecipitation assay.

Using a monoclonal antibody against the human nerve growth factor (NGF) receptor (20.4 IgG), a specific immunoprecipitation assay for the quantitation of human NGF binding sites has been established. The procedure involves specific labeling of NGF receptors by covalent crosslinking to 125I-NGF with a carbodiimide reagent, and subsequent immunoprecipitation of the detergent-extracted 125I-NGF-receptor complexes with the 20.4 IgG. This two-site assay provides a specific method to measure NGF binding sites in peripheral tissues and central nervous system. Moreover, it allows analysis of the immunoprecipitated receptor species by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The finding that specific NGF binding sites are expressed on human neuronal and nonneuronal tissues, including lymphoid tissues, indicates that NGF exerts a broader physiological function than originally ascribed.

Aged↗
PubMed1990

Nerve growth factor expression in the developing hippocampus isolated in vitro.

Nerve growth factor (NGF) is synthesized in the hippocampus and neocortex and provides trophic support for afferent cholinergic neurons of the basal forebrain. To determine the capacity of the developing hippocampus to express NGF in the absence of NGF-responsive afferents, embryonic hippocampal cells isolated prior to septal innervation were studied in reaggregating cell culture. The expression of NGF protein in vitro was qualitatively and quantitatively similar to that observed in situ. The expression of NGF mRNA exhibited an initial increase in vitro but then plateaued and was maintained at a steady level. This latter finding was in contrast to the steady rise in NGF mRNA levels observed in situ. These data suggest that (i) intrinsic hippocampal interactions regulate the onset of NGF expression, but that (ii) additional extrinsic developmental signals may be required for proper regulation of hippocampal NGF expression during ontogeny.

Aging↗
PubMed1989

Nerve growth factor increases in adult rat brain after hypoxic injury.

Brain injury increases the synthesis of many growth and trophic factors. We have measured nerve growth factor (NGF) content in brain of adult rats with bilateral carotid artery occlusion exposed to 7% O2. Five days after hypoxia the NGF content was increased in neostriatum, hippocampus and cerebral neocortex, areas exhibiting focal neuronal degeneration and inflammation. The increase was not related to changes in choline acetyltransferase activity that was only reduced in heavily damaged cortex. The findings may indicate that the increase in NGF content was due to enhanced NGF synthesis by inflammatory cells including stimulated glia.

Animals↗
PubMed1989

Long-term cultivation of cryopreserved human fetal brain cells in a chemically defined medium.

Conditions for long-term cultivation of human fetal brain cells in a chemically defined medium were established using cryopreserved brain fragments obtained from legal abortions. Tissue of the same gestational age was pooled and the cells cultured in a fully defined medium containing insulin-like growth factors (IGF I and II). Primary cultures were kept for 2-4 weeks and secondary or tertiary cultures could be maintained for 3 months. The cultures were characterized by morphological, electrophysiological and biochemical methods. Glial cells were predominant during the first two weeks of culture. In later stages of cultivation, glial cells diminished in number and most cells were neuronal. Voltage-dependent Na+ channels were recorded from neurons. Biochemical studies indicated that the fetal brain cells contained and secreted immunoreactive somatostatin as well as the tachykinins, substance P and neurokinin A. Cultures grown in IGF II- or nerve growth factor-containing medium expressed increased choline acetyltransferase activity.

Brain↗
PubMed1989

Developmental regulation of nerve growth factor and its receptor in the rat caudate-putamen.

In prior studies, nerve growth factor (NGF) administration induced a robust, selective increase in the neurochemical differentiation of caudate-putamen cholinergic neurons. In this study, expression of NGF and its receptor was examined to determine whether endogenous NGF might serve as a neurotrophic factor for these neurons. The temporal pattern of NGF gene expression and the levels of NGF mRNA and protein were distinct from those found in other brain regions. NGF and high-affinity NGF binding were present during cholinergic neurochemical differentiation and persisted into adult-hood. An increase in NGF binding during the third postnatal week was correlated with increasing choline acetyltransferase activity. The data are consistent with a role for endogenous NGF in the development and, possibly, the maintenance of caudate-putamen cholinergic neurons.

Animals↗
PubMed1989

Nerve growth factor induces growth and differentiation of human B lymphocytes.

Nerve growth factor (NGF) is known to affect peripheral sympathetic and sensory neurons as well as defined populations of neurons in the central nervous system. This paper presents evidence that NGF is also active in modulation of B-cell-mediated immune responses. NGF receptors were immunoprecipitated from highly purified human B-cell populations, and to a lesser extent, from T-cell populations, by using a monoclonal antibody recognizing NGF receptors present on neural cells. NGF receptors were also detected in significant amounts in human spleen and lymph node tissue. In addition, NGF induced a dose-dependent increase in B-cell DNA synthesis as determined by incorporation of [3H]thymidine. This B-cell growth-promoting activity was inhibited by a neutralizing anti-NGF monoclonal antibody. Immunoglobulin secretion, principally affecting IgM synthesis, was also modulated by NGF. The concentrations that affected B-cell proliferation are consistent with the presence of functional high-affinity NGF receptors. The results suggest that NGF, in addition to its neurotrophic function, also acts as an immunoregulatory cytokine.

Antibodies, Monoclonal↗