Biomedical subjects

C Grothe

Publications and source records attributed to C Grothe.

At least 37 records · Page 2Linked to original sources

PubMed1999

Localization and regulation of basic fibroblast growth factor (FGF-2) and FGF receptor-1 in rat superior cervical ganglion after axotomy.

In response to peripheral nerve lesion, synthesis of basic fibroblast growth factor (FGF-2) increases in sensory ganglia and motoneurons. Here, we investigated the axotomy-induced regulation of FGF-2 and FGF receptor-1 (FGFR-1) expression in the autonomic nervous system using the sympathetic superior cervical ganglion of the adult rat as a model. Transcripts for both proteins were detected by ribonuclease protection assay. Western blotting indicated the presence of all three FGF-2 isoforms (18, 21, and 23 kD) in the superior cervical ganglion. Immunohistochemical analysis revealed FGF-2 localization in nuclei of satellite cells surrounding postganglionic perikarya. After transection of the carotid nerves, the number of FGF-2-immunoreactive glial cells increased. FGF-2 mRNA was up-regulated within 6 h and remained elevated for 3 weeks. The 18-, 21-, and 23-kD isoforms were all increased 7 days after axotomy. FGFR-1 immunoreactivity was observed in neuronal and nonneuronal nuclei in the normal rat superior cervical ganglion. In contrast to FGF-2, expression of FGFR-1 was unchanged in ganglia after axotomy. Taken together, the present results suggest that FGF-2 participates in neuron-glial interactions of sympathetic ganglia and may be involved in sympathetic neuron survival or nerve regeneration after nerve lesion.

Animals↗

PubMed1998

Over-expression of the 18 kD and 21/23 kD fibroblast growth factor-2 isoforms in PC12 cells and Schwann cells results in altered cell morphology and growth.

Basic fibroblast growth factor (FGF-2) occurs in different isoforms which represent alternative translation products from a single mRNA. The question of whether the presence of multiple FGF-2 isoforms has physiological implications is compelling but unresolved so far. However, it has been shown recently that the FGF-2 isoforms are differentially regulated in sensory ganglia and peripheral nerve following nerve injury and, moreover, in the adrenal medulla during postnatal development and after hormonal stimuli suggesting that the isoforms may serve different physiological functions. To investigate isoform-specific effects we have established immortalized Schwann cells and PC12 cells stably over-expressing the 18 kD and the HMW isoforms. We found that the over-expression of the different isoforms alters morphology and growth of the Schwann cells. PC12 cells over-expressing the 18 kD FGF-2 were found to differentiate towards the neuronal phenotype whereas over-expression of the HMW isoforms resulted in a stabilization of the endocrine phenotype. Taken together, these data corroborate the idea of FGF-2 isoform-specific functions.

Animals↗

PubMed1998

Regulation of nerve growth factor and its low-affinity receptor (p75NTR) during myogenic differentiation.

In our preceding report, we have shown that nerve growth factor (NGF) and its low-affinity receptor (p75NTR) are expressed in C2C12 myoblasts and downregulated during myogenic differentiation. Furthermore, NGF affects myogenic differentiation and cell growth via p75NTR and downregulation of p75NTR is essential for myogenic differentiation (Seidl et al., 1998). Here we show that NGF and p75NTR are regulated by mechanisms preceding terminal differentiation in myogenic cells. These mechanisms include cell-density phenomena such as cell-cell contact as well as signaling of basic fibroblast growth factor (FGF-2) and its receptor (FGFR1). Downregulation of NGF and p75NTR occurred as a consequence of increasing cell density, an important trigger for the onset of myogenic differentiation. FGF-2 and FGFR1 were shown to be present in C2C12 cells and exogenous FGF-2 induced NGF and p75NTR expression, implying that FGF/FGFR signaling is an upstream regulator of the NGF/p75NTR system. The fact that FGF-2 could suspend yet not abolish density-induced downregulation indicates that cell-cell contact counteracts the FGF effect and ultimately terminates NGF/p75NTR signaling. This evidence, together with the observation that p75NTR expression is suppressed in muscle progenitors, which constitutively express adenovirus E1A proteins and thus lack the competence of myogenic differentiation, underline the important role for the NGF/p75NTR system in the interplay of multiple factors and biological systems that balance myogenic differentiation at the appropriate spatial and temporal level.

Adenovirus E1A Proteins↗

PubMed1998

Localization of fibroblast growth factor 2 (FGF-2) protein and the receptors FGFR 1-4 in normal human seminiferous epithelium.

Fibroblast growth factor 2 (FGF-2), which occurs in various isoforms both species and tissue specifically, regulates cell proliferation and differentiation via a dual receptor system consisting of heparan sulphate proteoglycans and receptor tyrosine kinases (FGFRs). This study demonstrates for the first time the distribution pattern of FGF-2 and the receptors FGFR 1-4 in the normal seminiferous epithelium of adult men. In western blot analyses, the polyclonal antibody, anti-FGF-2, shows two immunoreactive bands at 18 and 24 kDa. On paraffin sections, positive immunoreaction occurs within the cytoplasm of spermatogonia. The distribution pattern of the polyclonal anti-FGFR 1-4 antibodies is as follows: anti-FGFR-1 (one 68-kDa band) stains nuclei and cytoplasm of spermatogonia; anti-FGFR-3 (five bands at 68, 78, 105, 125 and 145 kDa) stains the nuclei of all germ cells except those of elongated spermatids; and anti-FGFR-4 (one 48-kDa band) stains the cytoplasm of primary pachytene spermatocytes. We were unable to demonstrate FGFR-2 immunoreactivity either in western blot analysis or on paraffin sections. This distribution pattern suggests that FGF-2 in spermatogonia is involved in the autocrine and paracrine regulation of the proliferation and differentiation of spermatogonia and spermatocytes via the receptors FGFR-1, FGFR-3 and FGFR-4.

Adult↗

PubMed1997

Expression of fibroblast growth factor-2 in hypoglossal motoneurons is stimulated by peripheral nerve injury.

We have studied the expression of basic fibroblast growth factor 2 (FGF-2) and FGF receptor 1 (FGFR1) in the hypoglossal motor system during degeneration and regeneration by using an RNase protection assay, in situ hybridization, and Western blot analysis. The FGF-2 transcript was found to be weakly expressed in the hypoglossal motoneurons of the adult rat. Both peripheral transection and crush injury of the hypoglossal nerve resulted in a marked up-regulation of the FGF-2 mRNA in motoneurons of the hypoglossal nucleus (with a peak at 10 and 11 days postlesion) as well as in the proximal and distal nerve stumps. The FGFR1 transcript was strongly expressed by hypoglossal motoneurons of unlesioned rats. Neither axotomy nor crush lesion of the hypoglossal nerve revealed any alteration of the expression level and cellular localization in the hypoglossal nucleus, but they did result in a significant increase of the FGFR1 mRNA level in the proximal and distal nerve stump. Western blot analysis of the hypoglossal nucleus revealed the presence of the 21 kD and 23 kD isoforms and of a weak expression of the 18 kD isoform. Hypoglossal nerve transection resulted in a complete down-regulation of the FGF-2 protein 3 days after lesion. After 14 days, however, the level of the three isoforms was increased above the control level. The regulation of FGF-2 in hypoglossal motoneurons after experimental nerve injury is in agreement with the idea of a lesion-related function of FGF-2. Together with previously reported neurotrophic effects, these results suggest that FGF-2 provides trophic support for lesioned motoneurons. At the injury site, FGF-2 could be involved in the regulation of the myelination.

Animals↗

PubMed1997

Differential expression of FGF-2 isoforms in the rat adrenal medulla during postnatal development in vivo.

Basic fibroblast growth factor (FGF-2) isoforms of the adrenal medulla are differentially expressed during rat postnatal development. While the 18 and 23 kDa isoforms continuously rise towards the adult expression level, the 21 kDa isoform displays a peak expression at postnatal day 28. The peak expression of the 21 kDa isoform correlates with the peak of the corticosterone concentration during postnatal development. Together with the previously demonstrated increase of the 21 kDa isoform in the adrenal medulla in vivo after glucocorticoid administration these results suggest that the differential regulation of the FGF-2 isoforms could be a physiologically occurring mechanism.

Adrenal Medulla↗

PubMed1997

Identification and characterization of differentiation-dependent Schwann cell surface antigens by novel monoclonal antibodies: introduction of a marker common to the non-myelin-forming phenotype.

In an attempt to identify and characterize novel Schwann cell surface molecules with putative functions during development, maintenance, and regeneration of the peripheral nervous system (PNS), we have produced monoclonal antibodies against viable neonatal rat Schwann cells. Using a sensitive live cell ELISA protocol, three monoclonal antibodies reactive with cultured Schwann cells, designated 27B10, 26F2, and 27C7 were isolated. The 27B10 and 26F2 antibodies specifically labelled forskolin-stimulated secondary Schwann cells in vitro as determined by live cell ELISA implying that the expression of the antigens in situ is regulated by axonal contact. The observation that the antigens seemed to be associated with both Schwann cell phenotypes clearly discriminated them from the well characterized myelin proteins as well as from molecules known to be confined to the non-myelin-forming phenotype. Interestingly, both antigens were found to be concentrated at the nodes of Ranvier. Further studies therefore have to show whether the identified antigens share structural or functional homology with adhesion or channel molecules, which display a similar distribution. Following transection of the adult sciatic nerve, the 26F2 antigen was rapidly down-regulated in the distal nerve stump. The 27C7 antibody reacted with an 80 kDa cell surface molecule common to non-myelin-forming Schwann cells. No differences in expression of the antigen between forskolin-treated and untreated Schwann cells in vitro were found, suggesting that the antigen is expressed independently from axonal contact. Two weeks after nerve transection in the absence of myelinating Schwann cells, the antigen was associated with S-100-positive Schwann cells of the distal nerve stump. The antigen was found to be expressed also by non-neuronal tissues, the level of the protein declined towards the adult stage. Comparison of the 27C7 antigen with previously described marker molecules suggests that we have identified a novel Schwann cell surface antigen of the non-myelin-forming phenotype.

Animals↗

PubMed1997

The multifunctionality of FGF-2 in the adrenal medulla.

Chromaffin cells of the adrenal medulla and their tumor counterparts, the pheochromocytoma (PC12) cells, are well-established model systems in neurobiology. The development of sympathoadrenal progenitor cells to chromaffin cells can be studied with regard to developmental signals which trigger the differentiation. With regard to potential treatments of neurological disorders like Parkinson's disease chromaffin cell grafting can be used as one therapeutical approach. The beneficial effect of chromaffin cell grafts is possibly not only related to the release of dopamine but may also be linked to the release of growth factors. One of the growth factors that is synthesized by chromaffin and PC12 cells is basic fibroblast growth factor (FGF-2). The experimental data available so far, are in agreement with different functional roles of FGF-2. This article summarizes the putative physiological functions of FGF-2 in the adrenal medulla. Three differential functional roles of FGF-2 are discussed: (1) as a differentiation factor for sympathoadrenal progenitor cells; (2) as a target-derived neurotrophic factor for preganglionic sympathetic neurons which innervate adrenal medullary cells; (3) as an auto-/paracrine factor in the adrenal medulla.

Adrenal Medulla↗

PubMed1997

Expression of fibroblast growth factor-2 and fibroblast growth factor receptor 1 messenger RNAs in spinal ganglia and sciatic nerve: regulation after peripheral nerve lesion.

In order to determine functional roles of basic fibroblast growth factor (FGF-2) in the peripheral nervous system we have analysed the expression of FGF-2 and FGF receptor 1 (FGFR1) in spinal ganglia and the sciatic nerve under normal conditions and after nerve crush using RNAse protection assay and in situ hybridization. In intact spinal ganglia, both FGF-2 and FGFR1 messenger RNAs are expressed, albeit at different levels. In situ hybridization identifies satellite cells as the source of FGF-2 and sensory neurons as the source of FGFR1 suggesting a paracrine mode of action of FGF-2 on sensory neurons. One day after crush lesion FGF-2 is significantly up-regulated in sensory ganglia L4-L6. Highest levels are found at day 7; control levels are approached after 28 days. FGFR1 messenger RNA, which is strongly expressed in intact spinal ganglia, displays no significant change after lesion. In the intact sciatic nerve, FGFR1 messenger RNA is detected at higher levels than FGF-2 messenger RNA. After injury, both transcripts display a time-dependent up-regulation in both the proximal and distal nerve stump. Schwann cells, as a putative source of the sciatic nerve-derived FGF-2, express both FGF-2 and FGFR1 messenger RNAs in vitro. The FGFR1 transcript level is increased in the presence of forskolin. FGF-2 does not affect expression of FGFR1 messenger RNA but stimulates its own expression. These results show that during peripheral nerve regeneration FGF-2 is up-regulated in both the crushed nerve and the respective spinal ganglia suggesting a possible physiological function of FGF-2 during the regeneration process.

Animals↗

PubMed1997

Differential regulation of fibroblast growth factor (FGF)-2 and FGF receptor 1 mRNAs and FGF-2 isoforms in spinal ganglia and sciatic nerve after peripheral nerve lesion.

To study the functional role of endogenous basic fibroblast growth factor-2 (FGF-2) during degeneration and regeneration of the sensory system, we have determined the expression and regulation of FGF-2 and FGF receptor (FGFR)-1 mRNAs in spinal ganglia and sciatic nerve during experimental transection and crush injury of the sciatic nerve. In contrast to levels of the FGFR-1 transcript, which is not altered, the level of FGF-2 mRNA is dramatically up-regulated in spinal ganglia after injury. In the proximal and distal nerve stumps both transcript levels are significantly elevated, albeit at different time points. The FGF-2 isoforms are differently up-regulated in spinal ganglia and sciatic nerve following peripheral nerve lesion. The differential response of FGF-2 mRNA and protein and of FGFR-1 mRNA in spinal ganglia and sciatic nerve after lesion is suggestive of different physiological functions: a local reaction at the lesion site where axonal regrowth occurs and a trophic reaction for the degenerating/regenerating sensory neurons.

Animals↗

PubMed1997

Localization, differential expression and retrograde axonal transport suggest physiological role of FGF-2 in spinal autonomic neurons of the rat.

Fibroblast growth factor-2 (FGF-2) has marked pharmacological neurotrophic effects on lesioned spinal autonomic neurons following target removal of the adrenal medulla, yet expression and axonal transport in autonomic neurons remain to be shown. We show here FGF-2 and FGF receptor type 1 (FGFR1) protein and mRNA expression in preganglionic intermediolateral neurons of the rat thoracic spinal cord. While immunoreactivity of both FGF-2 and FGFR1 co-localize to intermediolateral neurons, mRNA transcripts of FGFR1, but not of FGF-2, are detectable in intermediolateral preparations by RNase protection analysis, suggesting protein translocation in vivo. Unilateral microinjection of 125iodinated FGF-2 into the adrenal medulla (a major target of intermediolateral neurons) results in significant accumulation of specific radioactivity in thoracic spinal cord tissue, including the intermediolateral neurons, and the ipsilateral splanchnic nerve. Emulsion autoradiography demonstrated labelling over ipsilateral intermediolateral neurons only. Neuronal co-localization of FGF-2/FGFR1 protein, differential mRNA expression, specific retrograde axonal transport and the known neurotrophic actions in vivo, strongly suggest unique physiological roles of FGF-2 in the autonomic nervous system.

Adrenal Medulla↗

PubMed1996

In vivo and in vitro effect of glucocorticoids on fibroblast growth factor (FGF)-2 and FGF receptor 1 expression.

In order to clarify the physiological function of fibroblast growth factor (FGF-2) in the adrenal medulla the regulation of FGF-2 and FGF receptor 1 (FGFR1) was studied in vitro and in vivo in response to glucocorticoids. To assess the effects of glucocorticoids, in vivo extracts of adrenal medulla and adrenal cortex were analyzed by RNase protection assay and Western blot analysis. PC12 cells were chosen as a model system to study the effects of glucocorticoids in vitro. In PC12 cells, dexamethasone (DEX) was found to stimulate dramatically the expression of both FGF-2 mRNA and protein. Western blot analysis revealed that exclusively the 21-kDa FGF-2 isoform was enhanced. In contrast to the FGF-2 mRNA level FGFR1 was not affected by treatment with glucocorticoids. In vivo FGF-2 mRNA level and 21-kDa FGF-2 isoform level are significantly enhanced in the adrenal medulla 24 h after DEX injection. In vivo application of DEX leads to an increase of the medullary and cortical FGFR1 transcript levels. Glucocorticoid effects on FGF-2 expression were not found in adrenal cortex, heart, skeletal muscle, and kidney, respectively, in vivo and in L6 rat myoblasts in vitro. In addition to adrenal medullary cells glucocorticoids elevated the FGF-2 mRNA and protein level also in vivo in the brain and in vitro in immortalized Schwann cells. The present results suggest that the 21-kDa FGF-2 isoform mediates a physiological function specific for neuronal tissue which is modulated by glucocorticoids.

Adrenal Cortex↗

PubMed1996

Fibroblast growth factor receptor 1 in skeletal and heart muscle cells: expression during early avian development and regulation after notochord transplantation.

Basic fibroblast growth factor (bFGF, FGF-2) mediates several biological functions during embryonic development. With regard to skeletal muscle formation, it has been suggested that FGF-2 is involved in the growth and differentiation of myogenic precursor cells. To identify the FGF-responsive cells we studied the expression of FGF receptor type I (FGFR-1) during early embryonic development of the chick. FGFR-1 immunoreactivity is present at all stages examined (embryonic day [E] 2-E5). Expression of FGFR-1 is found in the somite myotome, limb bud muscle cells, eye and tongue muscle cells, and myocardium. Transplantation of an additional notochord into the paraxial mesoderm, which prevents the formation of a myotome, reveals the absence of FGFR-1 immunoreactivity on the operated side. The distinct expression pattern of FGFR-1 in migrating and differentiating muscle cells indicates that in addition to the stimulation of proliferation of myoblasts, FGF-2 exerts other (nonmitogenic) effects on postmitotic myocytes.

Animals↗

PubMed1996

A sensitive RNase protection assay using 33P labeled antisense riboprobes.

This article presents for the first time a modified protocol for RNase protection analysis that allows the substitution of 32P with 33P without loss of the high sensitivity of this method achieved with 32P. With this protocol, we were able to detect at least 1 pg of specific mRNA. In the RNase protection analysis 33P labeled riboprobes are more advantageous with regard to an easier handling and better resolution.

Animals↗

PubMed1996

Fibroblast growth factor receptor 1 in the adrenal gland and PC12 cells: developmental expression and regulation by extrinsic molecules.

In the present study we have analyzed the expression of fibroblast growth factor receptor 1 (FGFR-1) mRNA in the developing and adult rat adrenal gland and in PC12 cells under different culture conditions. For this purpose a sensitive ribonuclease protection assay using 33P-labelled riboprobes was established. 33P-labelled riboprobes show a high resolution and are relatively easy to handle. FGFR-1 mRNA was found to be present in the postnatal and adult adrenal gland. In the cortex high levels of FGFR-1 mRNA were detected at postnatal day (P) 1 and P8, during the third week the mRNA levels declined, and reached low levels during adulthood. PC12 cells also contained detectable amounts of FGFR-1 mRNA. With the exception of NGF, however, the different treatment procedures did not affect FGFR-1 mRNA levels. The expression pattern of the FGFR-1 transcript matches that of the expression of FGF-2 and of the mitotic activity in the developing and adult cortex. This supports the idea that FGF-2 might act as an autocrine mitogen for adrenocortical cells. In the medulla FGFR-1 mRNA levels were low at the first 3 postnatal weeks and increased towards the adult. In accordance with the developing expression pattern of FGF-2 in the medulla and in vitro effects of this protein on chromaffin and PC12 cells an autocrine/paracrine role as a maintenance and differentiation factor for chromaffin cells is conceivable.

Adrenal Glands↗

PubMed1996

Fibroblast growth factor and its implications for developing and regenerating neurons.

FGF is a multifunctional heparin-binding protein which was characterized by its mitogenic and angiogenic action outside the nervous system. Recent data confirm this multifunctionality also with regard to the nervous system. The distribution of FGF and its receptors seems not to be in agreement with the idea of a single function for one population but argues for a more complex action, which might be dependent on the development stage and cell type. FGF and its receptors are widely distributed in the nervous system. In brainstem and spinal cord motoneurons and in sensory ganglia the FGF-2 staining pattern is developmentally regulated suggesting a functional change during embryonic and postnatal development. In addition, after nerve lesion the FGF-2 expression is altered in sensory and motoneurons. Administration of FGF-2 reveals trophic effects on survival and transmitter metabolism in vivo and in vitro. According to a more general neurotrophic factor concept, a physiological role of FGF for distinct neuron populations during development is likely. In the motor system, for example, FGF could act synergistically with certain neurotrophins, CNTF, or other non-identified co-factors. In the sensory system, a possible non-neurotrophic role for at least postnatal and adult sensory neurons has to be further addressed in the future. In order to further define and characterize the actions of the FGFs a mapping of the different family members and their respective receptor molecules during development and in the adult has to be done.

Animals↗

PubMed1995

Fibroblast growth factor (FGF)-2 sense and antisense mRNA and FGF receptor type 1 mRNA are present in the embryonic and adult rat nervous system: specific detection by nuclease protection assay.

In the present study, we attempted to clarify the controversial question whether basic fibroblast growth factor (bFGF, FGF-2) mRNA is present or absent in the embryonic central nervous system (CNS). For this purpose we analyzed the expression of the FGF-2 mRNA in the embryonic and adult forebrain, brainstem, and spinal cord using the highly specific ribonuclease protection assay. Using this method we were able to detect FGF-2 mRNA in the rat CNS of embryonic day (E) 16 and 17, however, at lower levels compared to adult FGF-2 mRNA levels. In addition, we show that a FGF-2 antisense transcript is expressed in embryonic CNS tissue. Furthermore, using this method, we demonstrate FGF receptor 1 mRNA in the rat embryonic and adult CNS. The presence of FGF-2 and FGF receptor 1 suggests a physiological role for this growth factor during the development of the embryonic CNS.

Aging↗

PubMed1995

Fibroblast growth factor-2-like immunoreactivity in auditory brainstem nuclei of the developing and adult rat: correlation with onset and loss of hearing.

Fibroblast growth factor-2 (FGF-2; basic FGF) is widely distributed in the developing and adult brain and has numerous effects on cultured and lesioned neural cells. The physiological role of FGF-2 in the unlesioned nervous system, however, is still not understood. We have studied the distribution of FGF-2 in the developing, adult, and functionally impaired central auditory system of the rat using specific antibodies and peroxidase-antiperoxidase immunocytochemistry. FGF-2-like immunoreactivity (FGF-2-IR) occurred in neuronal cell bodies and/or nerve fibers but was very rarely observed in glial cells. Several auditory brainstem nuclei, including the superior paraolivary nucleus, the medial superior olive, the lateral and ventral trapezoid nuclei, and the central nucleus, as well as the external cortex of the inferior colliculus, were entirely devoid of FGF-2-IR. In the dorsal cochlear nucleus, the lateral superior olive, and the nuclei of the lateral lemniscus, FGF-2-IR was not detectable in nerve cell bodies prior to adult age. Neurons in the medial geniculate body exhibited FGF-2-IR only transiently, from postnatal day (P) 5 until P16. Neurons in the medial nucleus of the trapezoid body were immunoreactive from P8 onwards. FGF-2-IR in anteroventral and posteroventral cochlear neurons disappeared at P14, i.e., at the onset of hearing, but immunoreactivity returned after P21. A transient expression of FGF-2 around the time when hearing function commences was observed in the dorsal cortex of the inferior colliculus.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗