Localization Of Nerve Growth Factor Research Articles

Immunohistochemical localization of nerve growth factor, tropomyosin receptor kinase A, and p75 in the bone and articular cartilage of the mouse femur.

Sequestration of nerve growth factor (NGF) significantly attenuates skeletal pain in both animals and humans. However, relatively little is known about the specific cell types that express NGF or its cognate receptors tropomyosin receptor kinase A (TrkA) and p75 in the intact bone and articular cartilage. In the present study, antibodies raised against NGF, TrkA, and p75 (also known as CD271) were used to explore the expression of these antigens in the non-decalcified young mouse femur. In general, all three antigens displayed a remarkably restricted expression in bone and cartilage with less than 2% of all DAPI+ cells in the femur displaying expression of any one of the three antigens. Robust NGF immunoreactivity was found in mostly CD-31− blood vessel-associated cells, a small subset of CD-31+ endothelial cells, an unidentified group of cells located at the subchondral bone/articular cartilage interface, and a few isolated, single cells in the bone marrow. In contrast, p75 and TrkA were almost exclusively expressed by nerve fibers located nearby NGF+ blood vessels. The only non-neuronal expression of either p75 or TrkA in the femur was the expression of p75 by a subset of cells located in the deep and middle zone of the articular cartilage. Understanding the factors that tightly regulate the basal level of expression in normal bone and how the expression of NGF, TrkA, and p75 change in injury, disease, and aging may provide insights into novel therapies that can reduce skeletal pain and improve skeletal health.

Localization of nerve growth factor (NGF) receptors in the mitochondrial compartment: Characterization and putative role

Background The neurotrophin NGF receptors trkA and p75NTR are expressed in the central and peripheral nervous system as well as in non-neuronal tissues; originally described to localize to the plasma membrane, recent studies have suggested other intracellular localizations for both NGF receptors. Scope of review In order to determine whether NGF receptors localize to the mitochondrial compartment mitochondria isolated from human kidney, rat tissues and a human podocyte as cell line before and after differentiation were used. Major conclusions Our results demonstrate that NGF receptors are localized in the mitochondrial compartment of undifferentiated human podocytes and in all tissues analyzed including rat central nervous system. In mitochondria p75NTR, but not trkA, co-immunoprecipitates with the adenine nucleotide translocator (ANT) and the phosphodiesterase 4 isoform A5 (PDE4A5). Moreover, NGF, via trkA, protects isolated mitochondria of rat brain cortex from mitochondrial permeability transition induced by Ca 2+. General significance Although NGF receptors have been described as mainly citoplasmatic so far, we proved evidence of their expression at the mitochondrial level and their interaction with specific proteins. Our results demonstrating the expression of NGF receptors in the mitochondria provide new insights into the role of NGF at subcellular level, in different areas of the organism, including CNS.

Expression of Nerve Growth Factor and Its Receptors, TrkA and p75, in Porcine Ovaries

The cellular localization of nerve growth factor (NGF) and its receptors (TrkA, p75) was investigated during the estrous cycle in gilts. Also, the levels of expression of these factors in walls of tertiary follicles and corpora lutea (CLs) were determined using Western blot. The ovaries from days 3, 7, 16 and 20 of the cycle revealed the presence of NGF and its receptors in oocytes of secondary and tertiary follicles, follicular cells of primary and secondary follicles, thecal and granulosa cells of tertiary follicles and steroidogenic cells of CLs. In wall cells of primary follicles, NGF, TrkA and p75 staining was strongest on day 16, while in secondary follicles, only p75 was more intensely stained on day 16 and 20. In walls of small (to 3 mm in diameter) and medium (4-6 mm in diameter) follicles, NGF staining was lower on day 16, and the p75 reaction was strongest on day 20. On day 20, NGF staining in large follicles (7-10 mm in diameter) was higher than in smaller follicles. The levels of NGF and p75 in small and medium follicles were highest on day 20. The contents of NGF and TrkA in large follicles on day 20 were higher than in smaller follicles. NGF and TrkA contents in CLs were highest on day 7. Our study demonstrates that NGF, TrkA and p75 are expressed in the ovary during the estrous cycle in gilts. These results suggest that NGF and its receptors may be important for ovarian function in cycling gilts.

Nerve growth factor localization in the nasal mucosa of patients with persistent allergic rhinitis

Nerve growth factor localization in the nasal mucosa of patients with persistent allergic rhinitis

Nerve growth factor localization in the nasal mucosa of patients with persistent allergic rhinitis

Nerve growth factor (NGF) and NGF receptors have been shown to be expressed by structural and infiltrating inflammatory cells in the human allergic bronchial mucosa and conjunctiva. In the nose, a positive immunostaining for NGF was recently reported in biopsies of subjects undergoing surgery for refractory nasal obstruction. This study was aimed at studying by immunohistochemistry NGF expression and localization in the nasal mucosa from subjects with moderate/severe persistent allergic rhinitis and natural allergen exposure. Immunostaining for NGF, tryptase and eosinophil cationic protein was performed in human nasal turbinate sections of 25 patients affected by persistent allergic rhinitis and sensitization to Dermatophagoides pteronyssinus. NGF was consistently expressed in the epithelium and in the submucosa of allergic rhinitic subjects, preferentially localized in eosinophils and mast cells. A strong NGF immunostaining was found in mucous cells of the epithelial lining and in the submucosal glands. As previously shown for allergic asthma and allergic conjunctivitis, NGF is also detectable in the nasal mucosa of patients with persistent allergic rhinitis. The preferential NGF localization in mucous cells of the epithelial lining and in the submucosal glands suggests a possible role for NGF in modulating secretion in allergic rhinitis and possibly other allergic diseases.

Localization of nerve growth factor and its receptors in the human nasal mucosa

Nerve growth factor (NGF) is a pluripotent mediator, the levels of which are elevated in nasal lavage fluids of individuals with allergic rhinitis at baseline. Levels of NGF increase after allergen challenge. In the current study, we tested the hypotheses that mast cells are the main source of human nasal NGF, and that NGF can potentially affect mucosal elements other than nerves. Immunostaining with antibodies against NGF, tryptase, CD3, eosinophil cationic protein, and the high-affinity (tyrosine kinase A) and low-affinity (p75) receptors for NGF was performed by using human nasal turbinate sections. Double immunofluorescence staining demonstrated NGF in only 2% (median) (1.3, 2.3; 25th, 75th percentiles) of mast cells, 0.2% (0, 0.4) of T cells, but in 62.2% (56.5, 68) of activated eosinophils. With immunohistology, NGF expression was consistently strongest in the submucosal glands and lesser in the epithelial lining. Both high-affinity and low-affinity receptors for NGF were localized not only on nerves, as expected, but also on nasal epithelium, submucosal glands, and some interstitial cells, but not on vascular endothelium. This study demonstrates that submucosal glands, nasal epithelium, and eosinophils constitute the major sources of NGF in the human nasal mucosa. That NGF receptors are found in cells other than nerves supports the notion that the role of this neurotrophin is broader than simple modulation of the sensorineural system. The distribution of NGF and its receptors and its established release during allergic reactions suggest that this factor participates in the pathophysiology of allergic rhinitis.

Localization of nerve growth factor, neurotrophin-3, and glial cell line-derived neurotrophic factor in nestin-expressing reactive astrocytes in the caudate-putamen of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated C57/Bl mice

To address the hypothesis that reactive astrocytes in the basal ganglia of an animal model of Parkinson's disease serve neurotrophic roles, we studied the expression pattern of neurotrophic factors in the basal ganglia of C57/Bl mice that had been treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to induce the degeneration of nigral dopamine neurons and parkinsonism. MPTP induced significant neuronal degeneration in the substantia nigra pars compacta as detected with Fluoro-Jade B staining, and this was accompanied by an increase in nestin-expressing astrocytes within the caudate-putamen. The number of nestin-positive reactive astrocytes in the caudate-putamen peaked within 3-5 days following MPTP treatment and then declined progressively toward the basal level by 21 days after treatment. Immunofluorescence and confocal microscopy confirmed coexpression of nestin or Ki-67 (cell proliferation marker) in glial fibrillary acid protein-positive astrocytes in the caudate-putamen. Double immunolabeling further revealed immunoreactivities for nerve growth factor (NGF), neurotrophin-3 (NT3), and glial cell line-derived neurotrophic factor (GDNF) in nestin-positive reactive astrocytes. Semiquantification of data obtained from mice 5 days after MPTP injection indicated that the majority of nestin-expressing cells expressed NGF (92%), NT3 (90%), or GDNF (86%). Our results present novel evidence of neurotrophic features among reactive astrocytes in the dopamine-depleted striatum. These nestin-expressing reactive astrocytes may therefore play neurotrophic roles in neural remodeling of the basal ganglia in Parkinson's disease.

Expression of Nerve Growth Factor (NGF), and Its Receptors trkA and p75 in Ovaries of the Cyclic Golden Hamster (<i>Mesocricetus auratus</i>) and the Regulation of Their Production by Luteinizing Hormone

In the present study, changes in localization of nerve growth factor (NGF) and its receptors, trkA and p75 in the ovary were investigated during the estrous cycle of the golden hamster. The effect of LH surge on changes in localization of NGF, trkA and p75 in the ovary was also investigated. NGF and its receptors trkA and p75 were localized in oocytes, granulosa cells and theca cells of various stages of follicles throughout the estrous cycle. NGF and its two receptors were also present in numerous interstitial cells and luteal cells. The number of interstitial cells staining positively for NGF and its two receptors was greater in ovaries of day 1 (day 1=day of ovulation) than the other days during the estrous cycle. Treatment with the antiserum against luteinizing hormone releasing hormone (LHRH-AS) at 1100 h on day 4 completely blocked ovulation. There were few positive reactions for NGF and its two receptors in interstitial cells 24 hr after LHRH-AS injection. The effect of LHRH-AS treatment was blocked by a single injection of 10 IU human chorionic gonadotropin. The distinct widespread distribution of NGF and its two receptors in the ovary of golden hamsters suggest that NGF may be an important growth factor for regulation of ovarian function. Furthermore, the LH surge may be an important factor for inducing production of NGF and its two receptors in interstitial cells of the cyclic golden hamster.

Wide range of lineages of cells expressing nerve growth factor mRNA in the nerve lesions of patients with vasculitic neuropathy: An implication of endoneurial macrophage for nerve regeneration

In situ localization of nerve growth factor (NGF) mRNA was examined in the nerve lesions of patients with vasculitic neuropathy. Double labeling of in situ hybridization for NGF mRNA and immunohistochemistry for cell markers showed that NGF mRNA was expressed in a wide range of lineages of cells: Schwann cells, infiltrating macrophages, T cells and perivascular cells. Round-shaped macrophages with early-phase features expressed high levels of NGF mRNA, in contrast to late-phase polymorphic macrophages, which expressed low levels of NGF mRNA. NGF mRNA was also expressed universally in T cells with various cell surface markers. Epineurial macrophages surrounding vasculitic lesions and endoneurial T cells expressed high levels of NGF mRNA in the damaged nerves. Moreover, the extent of endoneurial NGF expression level in macrophages was closely related to the degree of axonal regeneration. These results suggest that NGF is expressed in a wide range of lineages of cells but is differentially expressed spatially in vasculitic nerve lesions, and that the expressed NGF, particularly in macrophages, may play an important role in the nerve regeneration process.

Neurotrophin and neurotrophin receptor protein expression in the human lung.

Neurotrophins (NTs) promote survival and differentiation of central and peripheral neurons, and display several activities also in non-neuronal cells. Human lungs synthesize and release NTs, which are probably involved in the pathophysiology of pulmonary disturbances. In this article the expression and anatomic localization of nerve growth factor, brain-derived neurotrophic factor, and NT-3 and of corresponding high-affinity receptors TrkA, TrkB (full-length and truncated [TR-] isoforms), TrkC, and of the low-affinity p75 receptor, were assessed in surgical samples from adult human lung by reverse transcriptase-polymerase chain reaction, Western blot, and immunohistochemistry. NTs and their cognate receptor mRNA and protein transcripts were detected by reverse transcriptase-polymerase chain reaction and immunoblotting, respectively, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) mRNA and corresponding protein transcripts being the most expressed. High levels of TrkB-[TR-] mRNA and of its protein transcript were also demonstrated, whereas a low expression of p75 mRNA and of corresponding protein transcript were found. Microanatomic analysis of immunohistochemical study revealed that bronchial epithelial cells were immunoreactive for different NTs, with a higher intensity of BDNF immune staining compared with other NTs, but did not express NT receptor immunoreactivity. Alveolar cells were immunoreactive for TrkA and TrkC receptor protein, but did not display immunoreactivity for NTs or other receptors investigated. Gland cells expressed NT and high-affinity NT receptor immunoreactivity, but not p75 receptor immunoreactivity. NT and low-affinity receptor immunoreactivity was observed within neurons and satellite cells of parasympathetic ganglia as well as in nerve fiber-like structures supplying the bronchopulmonary tree. An obvious immunoreactivity for NTs and NT receptor protein was also observed in intrapulmonary branches of pulmonary artery. Pulmonary lymphocytes and macrophages express nerve growth factor and high-affinity NT receptor immunoreactivity. The role of NTs in non-neuronal tissue including lung has not been clarified yet. The widespread expression of NTs and their receptors in different components of the lung suggests that these factors may contribute to regulate cell function in human lung.

Localization of nerve growth factor (NGF) receptors in the human nasal mucosa

Localization of nerve growth factor (NGF) receptors in the human nasal mucosa

Immunohistochemical Localization of Nerve Growth Factor, Glial Fibrillary Acidic Protein and Ciliary Neurotrophic Factor in Mesencephalon, Rhombencephalon, and Spinal Cord of Developing Mongolian Gerbil

The distribution of the nerve growth factor (NGF), the glial fibrillary acidic protein (GFAP) and the ciliary neurotrohic factor (CNTF) was performed in coronal sections of the mesencephalon, rhombencephalon and spinal cord in the developing Mongolian gerbils. Generally, NGF specifically recognizes neurons with the NGF receptor, whereas GFAP does the glia, and CNTF does the motor neurons. The receptor expression was examined separately in gerbils between embryonic days 15 (E15) and postnatal weeks 3 (PNW 3). The NGF-IR was first observed in the spinal cord at E21, which might be related to the maturation. The GFAP reactivity was peaked at the postnatal days 2 (PND2), while the highest CNTF-reaction was expressed at PNW 2. The GFAP stains were observed in the aqueduct and the spinal cord, which appeared to project laterally at E19. The CNTF was observed only after the birth and found in both the neurons and neuroglia of the substantia nigra, mesencephalon, cerebellum and the spinal cord from PND1 to PNW3. These results suggest that NGF, GFAP and CNTF are important for the development of the neurons and the neuroglia in the central nervous system at the late prenatal and postnatal stages.

Cellular Localization of Nerve Growth Factor (NGF) and Its Receptors TrkA and p75LNGFR in the Ovary of the Japanese Monkey (Macaca fuscata fuscata).

Cellular Localization of Nerve Growth Factor (NGF) and Its Receptors TrkA and p75LNGFR in the Ovary of the Japanese Monkey (Macaca fuscata fuscata).

Immunohistochemical localization of nerve growth factor in fractured and unfractured rat bone.

We detected nerve growth factor (NGF) by immunohistochemical localization in both fractured and unfractured rat rib. In unfractured bone, periosteal mesenchymal osteoprogenitor cells appeared to be the only skeletal cells which stained for NGF. Adjacent skeletal muscle fibers exhibited NGF staining both in fractured and unfractured bone. Fracture callus periosteal osteoprogenitor cells, marrow stromal cells, osteoblasts, young osteocytes and endothelial cells of new capillaries had moderate to heavy staining for NGF at 1 and 3 weeks after fracture. Deeply positioned osteocytes and osteoclasts showed no NGF staining. Most chondrocytes of fracture calluses stained for NGF, however, some chondrocytes did not stain which may indicate that NGF is produced at particular stages of chondrocytic differentiation. In calluses, periosteal matrix stained heavily for NGF when juxtaposed to cartilage and less obviously when associated with new bone at both 1 and 3 weeks post-fracture. However, other fibrous, cartilaginous and osseous matrices did not stain for NGF at any time. At 6 weeks post-fracture, NGF staining was largely confined to periosteal osteoprogenitor cells. The detection of NGF in periosteal osteoprogenitor cells of unfractured rib points to these cells having a role in nerve maintenance in intact bone. Furthermore, the localization of NGF in osteoprogenitor cells, marrow stromal cells, osteoblasts, certain chondrocytes, endothelial cells, periosteal matrix of the fracture callus and skeletal muscle may mean that these entities participate in fracture innervation. The presence of NGF in the callus may also indicate a direct, as yet undefined action of this neurotrophin on skeletal cell metabolism.

Localization of nerve growth factor, trkA and p75 immunoreactivity in the hippocampal formation and basal forebrain of adult rats

In the immunohistochemical staining of nerve growth factor, it has been reported that fixation-dependent lability of nerve growth factor hampers its localization. In the present study, we used two different polyclonal antibodies to immunostain nerve growth factor in rat brain tissue. We found that in paraformaldehyde-fixed (immersion- or perfusion-fixed) brains, nerve growth factor-like immunoreactivity was located primarily in the cytoplasmic membrane and fiber tract of hippocampal neurons and was sparse in cortical neurons. When fresh frozen brain sections were fixed in paraformaldehyde solution, nerve growth factor-like immunoreactivity was distributed evenly in the cell body. However, when fresh frozen brain sections were fixed in acetone, immunoreactivity to nerve growth factor was present as discrete or confluent dense particles in the cell body, especially in the nuclear region. Also, when paraformaldehyde-perfusion-fixed brain sections were heat treated in salt solution before immunostaining, nerve growth factor-like immunoreactivity could be retrieved in the cytoplasmic and nuclear regions. The hippocampal formation, cerebral cortex and basal forebrain expressed nerve growth factor-like immunoreactivity. Double immunostaining in fresh frozen brains showed that the low-affinity nerve growth factor receptor (p75) co-expressed with nerve growth factor and trkA proto-oncogene in basal forebrain neurons.Our study shows that formaldehyde fixation can mask nerve growth factor antigen, and special treatment, such as heating, is needed to retrieve nerve growth factor antigen to permit immunohistochemical detection. For immunohistochemical study of nerve growth factor in rat brain tissue, successful immunostaining can be obtained by using fresh frozen brains to prevent the masking effect of fixatives or by using paraformaldehyde-fixed brains with heat treatment. It is likely that nerve growth factor is synthesized and accumulated mainly in the cell body but not in the fiber tracts, which is similar to the distribution of its messenger RNA. The co-existence of p75 with nerve growth factor and trkA in basal forebrain neurons suggests the role of low- and high-affinity receptors in regulating the trophic effect of nerve growth factor.

Localization of nerve growth factor (NGF) and low‐affinity NGF receptors in touch domes and quantification of NGF mRNA in keratinocytes of adult rats

Touch domes are clearly delineated mechanoreceptors that are visible on the depilated skin of mammals. These structures consist of a sharply circumscribed disk of thickened epithelium surmounting a group of Merkel cells that are innervated by type I sensory neurons. These characteristic cutaneous structures provide an ideal opportunity for investigating whether the localization of nerve growth factor (NGF) in the skin is related to sites of sensory axon termination. For these reasons, we have used immunocytochemistry to study the distribution of NGF and the low-affinity NGF receptor (p75NGFR) in the touch domes of adult rat skin. Intense NGF-like immunoreactivity was sharply restricted to keratinocytes (excluding the stratum corneum) of the thickened epidermis of touch domes. The epidermis immediately surrounding touch domes and the epidermis of the tylotrich hair follicle associated with touch domes were not stained by anti-NGF antiserum. Merkel cells of the basal epidermis of touch domes were immunonegative for NGF but were immunopositive for p75NGFR as were the type I nerve endings innervating these cells. Quantitative Northern blotting revealed that the level of NGF mRNA was substantially higher in keratinocytes isolated from the stratum granulosum and stratum spinosum than in keratinocytes isolated from the stratum germinativum. These findings indicate that NGF synthesis in mature skin has a highly restricted regional distribution that is primarily associated with the innervation of a specialized touch receptor.

Intestinal localization of nerve growth factor in normal rats, mice, and megacolonic mice

Intestinal localization of nerve growth factor in normal rats, mice, and megacolonic mice

Immunohistochemical localization of nerve growth factor in the rat pineal gland

Sympathetic nerve fibers arising from the superior cervical ganglia are the main innervation of the rat pineal gland. Since most organs innervated by these ganglia contain nerve growth factor (NGF), the hypothetical existence of NGF in the pineal gland was investigated. The peroxidase anti-peroxidase technique was applied for the immunohistochemical demonstration of NGF using a polyclonal antiserum on Bouin-fixed, paraffin-embedded pineal glands from adult, young and 6-hydroxydopamine (6-OHDA)-treated rats. Few immunopositive cells were observed in the adult pineal gland. A more conspicuous population of immunoreactive cells displayed a stellate shape resembling the interstitial or glial cells previously described in the rat pineal gland. Since NGF plays a trophic effect on sympathetic neurons during development and adulthood, we postulate that its presence in the pineal gland may exert a trophic role on its sympathetic innervation.

Immunohistochemical localization of nerve growth factor (NGF) and NGF receptor (NGF-R) in the developing first molar tooth of the rat

Abstract Nerve growth factor (NGF) is a well established target-derived trophic factor supporting sympathetic and sensory innervation in the peripheral tissues as well as cholinergic innervation in the brain. Despite its name, NGF may have broader biological functions early in development in a wide range of non-neuronal differentiating cells. The many effects of NGF are directly dependent on initial binding of NGF to specific plasma membrane receptors on target cells. Here we use immunohistochemical methods to show that NGF and its receptor (NGF-R) are localized in a variety of embryonic epithelial and mesenchymal cells in the rat developing molar tooth. Dental cells known to play important roles in morphogenesis and inductive tissue interactions show NGF-like reactivity. Thus, labelling is seen in epithelial preameloblasts and mesenchymal odontoblasts. We also show a transient expression of NGF-R in restricted parts of the dental epithelium (inner dental epithelium) and dental mesenchyme differentiating cells (post-mitotic, polarizing odontoblasts). The expression patterns of NGF are different to those of NGF-R during embryogenesis and this is illustrated in detail in the developing tooth. The histochemical findings reported here support the notion that NGF may have multiple roles during morphogenetic and cytodifferentiation events in the tooth.

Nerve growth factor and serum differentially regulate development of the embryonic otic vesicle and cochleovestibular ganglion in vitro

The preceding paper ( P. Bernd and J. Represa, 1989, Dev. Biol. 134 ) describes the characterization and localization of nerve growth factor (NGF) receptors in inner ear primordia, the otic vesicle (OV) and cochleovestibular ganglion (CVG), obtained from 72-hr (stage 19–20) quail embryos. The studies described in this paper investigated whether NGF serves as a mitogen, a survival factor, and/or a differentiation factor in this system. Explants of isolated OV and CVG were maintained for 24 hr in serum-free medium alone (M-199), M-199 containing serum, M-199 containing NGF, or M-199 containing both serum and NGF. [ 3H]Thymidine was also present for the entire culture period. Both OV and CVG incorporated greater amounts of [ 3H]thymidine in the presence of serum or NGF, and their combined effect was additive. NGF's effects were dose dependent, saturable, and specific (blocked by anti-NGF). NGF caused little or no morphological differentiation of OV and no increase in protein levels, in contrast to OV grown in the presence of serum. CVG had both cochlear and vestibular portions present in all cases, but the apparent size and protein content of CVG was increased in the presence of either serum or NGF. Effects of serum and NGF were completely, but reversibly, blocked by amiloride, suggesting that the Na +H + exchange system had been activated. In order to determine whether increases in [ 3H]thymidine incorporation were due to increased cell survival or perhaps to an increase in proliferation, explants were initially grown for a 24-hr period in serum-free medium, followed by reactivation for an additional 24 hr in medium containing serum and/or NGF. It is likely that cells requiring either serum or NGF for survival would die during a 24-hr period in their absence. Our results revealed that the level of [ 3H]thymidine incorporation in OV was the same after reactivation. In the case of CVG, only NGF treatment yielded similar results; [ 3H]thymidine incorporation was lower in CVG reactivated with serum. It appears, therefore, that serum has probable proliferative effects upon OV and CVG, as well as survival effects for CVG. NGF, however, does not appear to affect survival in either OV or CVG, so that increases in [ 3H]thymidine incorporation in response to NGF are most likely due to proliferative effects upon OV or CVG, at least at this embryonic stage.