Fibroblast Growth Factor Immunoreactivity Research Articles

Effects of subcutaneous injection of ozone during wound healing in rats

Fibroblast growth factor 2 (FGF2) regulates the wound repair process and it is secreted by inflammatory and endothelial cells, and by myofibroblasts. This study aimed to establish the expression patterns of FGF2 and myofibroblastic differentiation during wound healing in rats treated with subcutaneous ozone injection. We created full-thickness excisional wounds in rats, and the healing process was analyzed through morphometric analyses and digital quantification of immunoreactivity of smooth muscle actin and FGF2. Ozone therapy-treated wounds presented granulation tissue with a reduced number of inflammatory cells and greater dermal cellularity, and intense collagen deposition. FGF2 immunoreactivity, microvessel density, and amount of myofibroblasts were significantly higher in treated wounds compared to controls. In conclusion, it was demonstrated that subcutaneous injections of ozone accelerate and ameliorate wound repairing process. Moreover, injectable ozone therapy’s action mechanism may be associated with FGF2 overexpression.

The effect of organic silicon injection on Achilles tendon healing in rats.

The aim of this study was to assess the efficacy of a soluble absorbable silicon compound on healing of the Achilles tendon. The Achilles tendons of 21 Wistar albino rats were cut and repaired. A 0.01 ml organic silicon solution (silanol) was injected peri/intratendinously into the left leg of all rats and the same dose of saline into the right leg postoperatively. Rats were randomly divided into 3 groups for biomechanical testing on Day 10 (7 rats) and Day 20 (7 rats) and histological and immunohistochemical assessment on Day 20 (7 rats). Fibroblast cell count and diameter, tissue vascularity and blood vessel diameter were evaluated by histomorphometry. Basic fibroblast growth factor (bFGF) immunoreactivity was analyzed with immunohistochemistry on Day 20. Failure load and stiffness of the repaired tendons were measured on Days 10 and 20. The number of fibroblasts per area, average fibroblast diameter, number of vessels parallel to collagen bundles per area and average vessel diameter were significantly higher in the organic silicon group than in the control group (p<0.05). Strong immunoreactivity of bFGF in the silicon group was detected. Failure load was significantly higher in the silicon group than in the control group on Day 10 (p=0.041). On Day 20, while a difference still existed, this difference was not significant. There was no effect of the silicon injection on stiffness of healing tendons. Organic silicon appears to have a positive effect on tendon healing and is suitable for further studies on host healing response modification.

Serotonin-Producing Enterochromaffin Cell Tumors of the Pancreas

Serotonin-producing tumors of the pancreas are rare endocrine neoplasms composed of enterochromaffin (EC) cells that have been mainly described in the literature as case reports. This study analyzes the clinicopathologic features of a series of pancreatic EC cell neoplasms and their similarities to and differences from intestinal EC cell tumors. The morphological, immunohistochemical, ultrastructural, and fluorescent in situ hybridization features of 15 pancreatic and 20 intestinal serotonin-producing neoplasms were compared. In addition, we reviewed the literature on pancreatic serotonin-producing tumors to better understand the clinicopathologic features of this rare tumor type. The lack of substance P and acidic fibroblast growth factor immunoreactivity; the low immunohistochemical expression of CDX2, vesicular monoamine transporter 1, connective tissue growth factor, and prostatic acid phosphatase; the lack of S100-positive sustentacular cells; the strong expression of vesicular monoamine transporter 2; and peculiar ultrastructural features characterize pancreatic EC cell tumors and differentiate them from intestinal ones, although both categories show similar chromosome 18 cytogenetic alterations. The review of the literature indicates that pancreatic functioning tumors associated with the carcinoid syndrome arise in younger patients and are larger, more frequently malignant, and more aggressive neoplasms than pancreatic nonfunctioning ones. Pancreatic EC cell tumors show several different morphological features compared with related intestinal tumors despite similar cytogenetic alterations on chromosome 18.

Human chorionic gonadotrophin regulates FGF2 and other cytokines produced by human endometrial epithelial cells, providing a mechanism for enhancing endometrial receptivity

Preimplantation cross-talk between a functional blastocyst and the endometrium is critical for successful blastocyst implantation. This interaction is mediated in part by endometrial cytokines/growth factors secreted by glandular epithelium into the uterine cavity. Recent evidence suggests that blastocyst-derived hCG may influence the endometrial milieu in conception cycles thereby enhancing receptivity and implantation success. This study investigated the effect of hCG on the secretory profile of a select cohort of 44 cytokines/growth factors from primary human endometrial epithelial cells (hEECs). These factors included those with both known and unknown roles during receptivity and implantation. The expression of one previously unknown hCG-regulated factor, fibroblast growth factor 2 (FGF2), in human endometrium and its effects on hEEC function were further examined. hEECs isolated from endometrial biopsies collected from fertile cycling women (n = 15) were treated ± recombinant hCG (0.2-20 IU/ml) for 48 h and conditioned media was quantitatively analysed using Luminex™ multiplex technology. FGF2 was further investigated by immunohistochemistry, western blot and cell-adhesion assays. Of 44 cytokines/growth factors examined, 39 were produced by hEECs with a distinct profile. hCG (2 IU/ml) significantly increased the production of six factors, including those with known roles in receptivity and trophoblast function (interleukin-11), blastocyst migration and adhesion (CXCL10), blastocyst development (granulocyte macrophage colony-stimulating factor) and one unknown with respect to receptivity and implantation (FGF2). Up-regulation of known hCG-regulated proteins, vascular endothelial growth factor and leukaemia inhibitory factor, validated this study. Immunoreactive epithelial FGF2 increased across the menstrual cycle, being highest in secretory and first trimester pregnancy endometrium in vivo. FGF2 (100 ng/ml) stimulated phosphorylation of ERK1/2 in hEEC with no effect on ERK1/2 abundance and stimulated hEEC adhesion to fibronectin and collagen IV (components of blastocyst/trophectoderm extracellular matrix). These findings clearly support roles for hCG and FGF2 in the blastocyst-endometrial cross-talk important for endometrial receptivity and blastocyst implantation.

FGF-2 in Astroglial Cells during Vertebrate Spinal Cord Recovery

Event Abstract Back to Event FGF-2 in Astroglial Cells during Vertebrate Spinal Cord Recovery Gehan Fahmy1 and Marie Moftah1* 1 Alexandria University, Zoology Department, Faculty of Science, Egypt FGF-2 is a pleiotrophic cytokine with neurotrophic and gliogenic properties both in vivo and in vitro. In the brain, it is localized in astrocytes and discrete neuronal populations. FGF-2 is known to regulate CNS injury responses, which include transformation of reactive astrocytes, scar formation, neurogenesis, and promotion of neurotrophic activities. Astrocytes become reactive following both central and peripheral nervous system injury. These activated cells undergo hypertrophy. A key indicator of astrocyte activation is the increased accumulation of intermediate filaments composed of glial fibrillary acidic protein (GFAP). Following a physical insult to brain or spinal cord, reactive astrocytes near the damage site show increased FGF-2 immunoreactivity. Thus, FGF-2 appears to participate in astrocytic differentiation and proliferation and a good candidate for astrocytic function regulation in healthy, injured, or diseased CNS. To further investigate the cellular mechanisms underlying FGF-2 restorative actions and to analyze, in more details, the changes within astroglial cells, we studied the localization of GFAP and FGF-2 in adult intact and injured Pleurodeles CNS. Our results show that spinal cord injury triggers a significant increase in FGF-2 immunoreactivity in reactive astrocytes at sites of insult. In addition, these results were time-dependent. Increase in FGF-2 immunoreactivity along the CNS axis, starting 1-week post-injury, was long-lasting extending to 6 weeks. This increase was accompanied by an increase in GFAP immunoreactivity in the same spatial pattern except in SC3 where its level was almost similar to sham-operated animals. Therefore, we suggest that FGF2 may be involved in cell proliferation and/or astroglial cells differentiation after body spinal cord transection in salamander, and could thus play an important role in locomotion recovery after spinal lesion.Acknowledgments: Part of the experiments were performed in the physiopathology of neuronal networks laboratory, INSERM U862, Equipe NAGY, Neurocentre Magendie, 146 rue Leo Saignat, 33077 Bordeaux Cedex, France. Keywords: Fibroblast growth factor-2 (FGF-2), Nerve Regeneration, Pleurodele waltlii Conference: 3rd Mediterranean Conference of Neuroscience , Alexandria, Egypt, 13 Dec - 16 Dec, 2009. Presentation Type: Poster Presentation Topic: CNS Diseases Citation: Fahmy G and Moftah M (2009). FGF-2 in Astroglial Cells during Vertebrate Spinal Cord Recovery. Front. Neurosci. Conference Abstract: 3rd Mediterranean Conference of Neuroscience . doi: 10.3389/conf.neuro.01.2009.16.151 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 25 Nov 2009; Published Online: 25 Nov 2009. * Correspondence: Marie Moftah, Alexandria University, Zoology Department, Faculty of Science, Alexandria, Egypt, marie.moftah@alexu.edu.eg Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Gehan Fahmy Marie Moftah Google Gehan Fahmy Marie Moftah Google Scholar Gehan Fahmy Marie Moftah PubMed Gehan Fahmy Marie Moftah Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Chronic antidepressant treatments increase basic fibroblast growth factor and fibroblast growth factor-binding protein in neurons

One of the mechanisms proposed for antidepressant drugs is the enhancement of synaptic connections and plasticity in the hippocampus and cerebral cortex. Fibroblast growth factor 2 (FGF2) is a growth factor essential for the proper formation of synaptic connections in the cerebral cortex, maturation and survival of catecholamine neurons, and neurogenesis. In this report, we attempted to establish a correlation between antidepressant treatments and FGF2 expression in the cerebral cortex and hippocampus, two brain areas relevant for depression. Desipramine (DMI, 10 mg/kg) or fluoxetine (FLU, 5 mg/kg) was injected acutely (single injection) or chronically (daily injection for two weeks) in adult rats. Chronic, but not acute, antidepressant treatments increase FGF2 immunoreactivity in neurons of the cerebral cortex and in both astrocytes and neurons of the hippocampus. FGF2 immunoreactivity in the cortex was increased mainly in the cytoplasm of neurons of layer V. Western blot analyses of nuclear and cytosolic extracts from the cortex revealed that both antidepressants increase FGF2 isoforms in the cytosolic extracts and decrease accumulation of FGF2 immunoreactivity in the nucleus. To characterize the anatomical and cellular specificity of antidepressants, we examined FGF-binding protein (FBP), a secreted protein that acts as an extracellular chaperone for FGF2 and enhances its activity. DMI and FLU increased FBP immunoreactivity in both cortical and hippocampal neurons. Our data suggest that FGF2 and FBP may participate in the plastic responses underlying the clinical efficacy of antidepressants.

Immunohistochemical expression of fibroblast growth factor (FGF)‐2 in epilepsy‐associated malformations of cortical development (MCDs)

To elucidate the biological significance of dysplastic cells in malformations of cortical development, an immunohistochemical study was performed to investigate fibroblast growth factor-2 (FGF-2) expression in corticectomy specimens from epilepsy patients, including focal cortical dysplasia (FCD) with balloon cells (BCs) (n=4; age/sex=2M, 14F, 24M, 45M), tubers of tuberous sclerosis complex (TSC-tubers) (n=2; 1F, 3F), FCD without BCs (n=3; 23F, 23M, 25M), and gliotic lesions (n=3; 12M, 25M, 29M). The nucleus and/or cytoplasm of astrocytes in all cases examined were positive for FGF-2; however, FGF-2 immunoreactivity was not detected in oligodendroglial cells. In all dysplastic lesions, FGF-2 was detected in the astrocytic nuclei, and cytoplasm and/or nuclei of BCs. Dysplastic neurons (DNs) in FCD with BCs and TSC-tubers were variably positive for FGF-2 in the cytoplasm, but FGF-2 was not detected in the neurons of FCD without BCs. The number of FGF-2 immunoreactive cells (FGF-2-IR%) in FCD with BCs (46.0+/-4.1%) was higher than that in FCD without BCs (19.8+/-3.1%) and gliotic lesions (19.5+/-3.3%) with statistical significance (P<0.001). These results, together with previous reports showing FGF-2 expression in neuroblasts and glioblasts in human fetal brain, and mainly in astrocytes in adult brain, suggest that FGF-2 expression in MCDs reflects incomplete differentiation and maturation of dysplastic cells, and that FGF-2-IR% is associated with histological subtypes of MCD, reflecting the timing of insults underlying the pathogenesis of each disorder.

Aortic coarctation hypertension induces fibroblast growth factor-2 immunoreactivity in the stimulated nucleus tractus solitarii

The actions of neurotrophic factors i.e. basic fibroblast growth factor (bFGF, FGF-2) to neurons are related not only to neuronal development and maintenance but also to synaptic plasticity regarding neurotransmission. We analyzed here the levels of FGF-2 immunoreactivity in the nucleus tractus solitarii (NTS) of Wistar Kyoto rats in response to alterations of neuronal activity promoted by the stimulation of the baroreceptor reflex following an aortic coarctation-induced-hypertension. The FGF-2 immunoreactivity (IR) was found in the cytoplasm of the neurons and in the nuclei of the glial cells in the NTS. A large number of NTS neurons expressed FOS immunoreactivity 4 h after coarctation, as an indication of neuronal activity. Stereological methods showed an increased number of FGF-2 immunoreactive (ir) neuronal profiles (90%) and glial profiles (149%) in the NTS of the 72 h aortic coarctated rats. 1-week later, FGF-2 ir neurons were still increased (54%) but no change was found in the number of FGF-2 ir glial profiles. The double immunoperoxidase method revealed that the majority of the FGF-2 ir glial cells was glial fibrillary acidic protein (GFAP) positive astrocytes. GFAP immunohistochemistry showed an astroglial reaction at 72 h time-interval (55%) but not 1 week after stimulation. The number of the cresyl violet positive neurons and OX42 ir profiles (marker of activated microglia) in the NTS of coarctated rats were not different from control by 1 week and 1 month after the surgery, indicating a lack of NTS injury in this period following coarctation hypertension. FGF-2 may be an important neurotrophic factor in areas involved in the control of blood pressure. The increased FGF-2 IR in the NTS cells following neuronal stimulation may represent trophic and plastic adaptive responses in this nucleus in an autocrine/paracrine fashion.

The fibroblast growth factor-2 antisense gene inhibits nuclear accumulation of FGF-2 and delays cell cycle progression in C6 glioma cells

Fibroblast growth factor-2 (FGF-2) is a potent heparin-binding protein with growth-promoting and anti-apoptotic activity. Transcription of the GFG/NUDT6 gene on the opposite DNA strand generates an overlapping antisense RNA (FGF-AS) implicated in the post-transcriptional regulation of FGF-2. C6 glioma cells coordinately express FGF-2 and FGF-AS mRNA in a cell cycle-dependent manner. Cellular FGF-2 immunoreactivity was also cell cycle-dependent, with marked nuclear accumulation during S-phase. Stable transfection and overexpression of the FGF-AS RNA resulted in suppression of total cellular FGF-2, and a reduction in nuclear accumulation of FGF-2 isoforms. Serum stimulation of growth-arrested wild-type cells evoked a rapid nuclear translocation of FGF-2, and cell cycle re-entry. FGF-AS transfectants, in contrast, showed a significant delay in recovery of both nuclear FGF-2 staining and S-phase re-entry. Similar results were observed when cells were released from aphidicolin-induced G1 arrest or subjected to heat shock. These findings indicate that FGF-AS RNA inhibits expression and cell cycle-dependent nuclear accumulation of FGF-2, and this is associated with a marked delay in S-phase progression. The results suggest that the endogenous FGF antisense RNA may play a significant functional role in the regulation of FGF-2 dependent cell proliferation in FGF-2 expressing cells.

Pattern of FGF-2 isoform expression correlated with its biological action in experimental prolactinomas

Fibroblast growth factor-2 (FGF-2) synthesized in the pituitary is involved in the formation and progression of pituitary tumors. The aim of this study was to analyze the pattern expression of two FGF-2 isoforms at different subcellular levels and to determine its correlation with prolactinoma development. Estrogen administration to male rats for 7, 20, and 60 days generated pituitary tumors, with lactotrophs being the prevalent cell type. Ultrastructural immunolabeling showed FGF-2 in the cytosolic and nuclear compartments of somatotrophs, lactotrophs and gonadotrophs, as well as in folliculo-stellate cells of normal rats. Estrogen stimulation increased FGF-2 immunoreactivity in various tumors and enhanced the expression of two FGF-2 isoforms, 18 and 22 kDa, as quantified by western blot. The 18 kDa isoform observed in cytosol extracts reached the highest levels after 60 days of hormonal stimulation and this was related to lactotroph proliferation. However, the 22 kDa FGF-2 isoform was only detected in the nuclear compartment and achieved the maximum expression at 7 days of estrogen treatment, without any correlation with lactotroph proliferation. These results suggest that the 18 kDa FGF-2 may play a role in the modulation of lactotroph proliferation in prolactinomas induced by estrogen. The overproduction of both FGF-2 isoforms appears to be implicated in autocrine-paracrine-intracrine mitogenic loops; this FGF-2 activity could lead to uncontrolled cell growth, angiogenesis, and tumor formation.

Apomorphine‐induced activation of dopamine receptors modulates FGF‐2 expression in astrocytic cultures and promotes survival of dopaminergic neurons

Apomorphine (APO), a potent D1/D2 dopamine receptor agonist, is currently used as an antiparkinsonian drug. We have shown previously that APO stimulates synthesis and release of multiple trophic factors, such as brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF), in both mesencephalic and striatal neurons, thereby effectively preventing dopaminergic neuron loss in vitro. The present study was designed to investigate the effects of APO on fibroblast growth factor-2 (FGF-2) expression and regulation in astrocytes, and furthermore, to identify signaling mechanisms underlying these effects. Here, we show that FGF-2 expression is robustly induced in cultured astrocytes in response to APO. FGF-2 expression was proportional to APO concentration and time-dependent. Conversely, treatment with S-APO, a derivative of R-APO lacking DA receptor agonist activity, did not alter FGF-2 levels. APO treatment resulted in enhanced cytosol FGF-2 immunoreactivity, export of high MW forms of FGF-2 to the cytoplasm from the nucleus and increased extracellular release of FGF-2. Interestingly, both high and low MW forms of FGF-2 were detectable in conditioned medium of APO-treated cultures. This APO-induced effect was correlated with activation of D1 and D2 receptors, as it could be either mimicked by dopamine receptor agonists (SKF38393, quinpirole) or partially blocked by antagonists (SCH23390, SKF83566, haloperidol). Activation of the D1 receptor preferentially increased PKA activity, whereas activation of the D2 receptor only promoted phosphorylation of MAPK. Importantly, APO-modulated FGF-2 expression was independent of Akt/phosphoinositide 3-kinase signaling. These data suggest that APO can enhance biosynthesis and release of FGF-2 through activation of dopamine receptors in striatal astrocytes. Both cAMP/PKA and MEK/MAPK signaling cascades are major steps mediating this process.

Fibroblast Growth Factor-2 Is Expressed by the Bovine Uterus and Stimulates Interferon-τ Production in Bovine Trophectoderm

Uterine-derived factors are essential for conceptus development and secretion of the maternal recognition-of-pregnancy factor, interferon-tau (IFNT), in ruminant species. The objectives of this study were to determine whether fibroblast growth factor-2 (FGF-2) is expressed in the bovine uterus during early pregnancy in cattle and to determine whether FGF-2 supplementation affects IFNT mRNA and protein abundance in bovine trophectoderm. FGF-2 mRNA was present in endometrium throughout the estrous cycle and was localized to the luminal and glandular endometrial epithelium at d 17-18 after estrus in pregnant and nonpregnant cows. Immunoreactive FGF-2 protein was detected within the endometrium and in the uterine lumen at d 17-18 after estrus, and concentrations did not differ based on pregnancy status. In a bovine trophectoderm cell line, CT-1, supplementation of medium with at least 1 ng/ml FGF-2 increased the incorporation of [(3)H]thymidine into DNA. Similarly, IFNT secretion from CT-1 cells increased after FGF-2 supplementation (1-100 ng/ml) for 72 h. Abundance of IFNT mRNA in CT-1 cells increased after 24 h exposure to 1, 10, or 100 ng/ml FGF-2. In bovine blastocysts, FGF-2 supplementation did not affect cell number after 72 h of culture but did stimulate IFNT protein concentrations in conditioned medium. In summary, FGF-2 is present in the uterine lumen during early pregnancy and increases IFNT mRNA and protein abundance in trophectoderm. The magnitude by which FGF-2 stimulates IFNT expression suggests that this uterine-derived factor plays an active role in regulating the establishment and maintenance of pregnancy in ruminants.

Expression and possible role of hPTTG1/securin in cutaneous malignant melanoma

Human pituitary tumour-transforming gene 1 or hPTTG1 is a proto-oncogene that codes for securin, a protein involved in sister chromatid separation. Based on previous microarray data, we studied the expression of hPTTG1/securin in melanocytic lesions. In contrast to nevi and radial growth phase melanomas, securin was expressed by scattered cells in the vertical growth phase, suggesting a role in tumour progression. In a series of 29 nodular and 29 superficial spreading melanomas, matched for all histological prognostic parameters, securin expression was significantly correlated with the nodular subtype (P=0.018) and not related to thickness. In other cancers, hPTTG1 is involved in various oncogenic pathways, including induction of neovascularisation and aneuploidy, and inhibition of p53 activity. We found coexpression of securin with wild-type p53 in the same neoplastic cells in a minority of melanomas. Expression of securin was significantly correlated with the extent of aneuploidy but not with basic fibroblast growth factor immunoreactivity or microvessel density. DNA cytometry revealed that nuclei-overexpressing securin frequently showed tetraploidy or aneuploidy. Our data show that hPTTG1 is frequently overexpressed in nodular melanoma, and suggest that hPTTG1 may act as an oncogene in the vertical growth phase, either by inhibiting anaphase, thereby causing aneuploidy and genomic instability, or by modulating the function of p53, thereby impairing apoptosis.

Anti-Fibroblast Growth Factor-2 Antibodies Attenuate Mechanical Allodynia in a Rat Model of Neuropathic Pain

Peripheral nerve injury leads to the activation of spinal cord astrocytes, which contribute to maintaining neuropathic (NP) pain behavior. Fibroblast growth factor-2 (FGF-2), a neurotrophic and gliogenic factor, is upregulated by spinal cord astrocytes in response to ligation of spinal nerves L5 and L6 (spinal nerve ligation [SpNL]). To evaluate the contribution of spinal astroglial FGF-2 to mechanical allodynia following SpNL, neutralizing antibodies to FGF-2 were injected intrathecally. Administration of 18 microg of anti-FGF-2 antibodies attenuated mechanical allodynia at day 21 after SpNL and reduced FGF-2 and glial acidic fibrillary protein mRNA expression and immunoreactivity in the L5 spinal cord segment of rats with SpNL. These results suggest that endogenous astroglial FGF-2 contributes to maintaining NP tactile allodynia associated with reactivity of spinal cord astrocytes and that inhibition of spinal FGF-2 ameliorates NP pain signs.

Differential effects of reproductive and hormonal state on basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in the hypothalamus and cingulate cortex of female rats

Morphological changes in astrocytes occur in a number of brain regions including the hypothalamus and hippocampal regions as a function of hormonal and reproductive state. Because basic fibroblast growth factor has been shown to play an important role in morphological changes in astrocytes, we investigated whether basic fibroblast growth factor immunoreactivity would also be influenced by reproductive state and circulating gonadal steroids. To do this we compared astrocytic basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in hypothalamic nuclei and the cingulate cortex, area 2 among groups of cycling, late pregnant and lactating rats as well as in ovariectomized and ovariectomized hormone-replaced females. Significant differences in both basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity were observed across groups in the supraoptic nucleus, parvocellular paraventricular nucleus, medial preoptic area of the hypothalamus and cingulate cortex 2. The pattern of change in basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity varied across regions both in direction and magnitude. For example, although in the supraoptic nucleus ovariectomized rats had lower levels of basic fibroblast growth factor-ir than cycling females, this pattern was reversed within cingulate cortex. Overall the results of this study suggest that reproductive and hormonal states are associated with robust changes in basic fibroblast growth factor and glial fibrillary acid protein immunoreactivity in a number of brain areas but that the changes observed vary in magnitude as well as direction from one brain region to another.

Early Inducible Nitric Oxide Synthase 2 (NOS 2) Activity Enhances Ischaemic Skin Flap Survival

A functional skin-flap model of angiogenesis in the mouse was utilized to investigate ischaemic flap survival/angiogenesis whilst under pharmacological or genetic inhibition of nitric oxide synthase (NOS). In this model, the epigastric artery was cauterized. Following a five-day angiogenic period an abdominal skin-flap supplied by the pre-existing epigastric artery was raised and resutured. After a further six days the outcome was determined by measuring the area of living skin-flap that was sustained by new vessel growth around the cauterized artery. Both pharmacological [ S-methyl-isothiourea (SMT) given via i.p. injection for the five-day angiogenic period] and genetic inhibition of NOS 2 (using NOS 2-/- mice) caused a similar and significant fall in flap survival compared to saline-treated wild-type mice ( P < 0.05). Delaying pharmacological NOS 2 inhibition for two days post-arterial cauterization increased flap survival in wild-type mice to that of saline-treated controls, whilst treating wild-type mice with SMT for only the first three days of angiogenesis produced a significant decrease in flap survival, similar to that of five-day SMT-treated wild types. Immunoreactivity for NOS 2 in NOS 2-/- knockout mice was absent in both pedicles and skin-flaps, whilst mast cells from both the pedicle and the skin-flap stained positively for NOS 2 in wild-type mice. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF 2) immunoreactivity was also strongly evident in mast cells of both wild-type and NOS 2-/- mice at both sites. These results point to a significant role for NOS 2 in promoting wound healing/angiogenesis in its early stages.

Atypical development of the tracheal basement membrane zone of infant rhesus monkeys exposed to ozone and allergen

Development of the basement membrane zone (BMZ) occurs postnatally in the rhesus monkey. The purpose of this study was to determine whether house dust mite allergen (HDMA) plus ozone altered this process. Rhesus monkeys were exposed to a regimen of HDMA and/or ozone or filtered air for 6 mo. To detect structural changes in the BMZ, we measured immunoreactivity of collagen I. To detect functional changes in the BMZ, we measured perlecan and fibroblast growth factor-2 (FGF-2). We also measured components of the FGF-2 ternary signaling complex [fibroblast growth factor receptor-1 (FGFR-1) and syndecan-4]. The width of the BMZ was irregular in the ozone groups, suggesting atypical development of the BMZ. Perlecan was also absent from the BMZ. In the absence of perlecan, FGF-2 was not bound to the BMZ. However, FGF-2 immunoreactivity was present in basal cells, the lateral intercellular space (LIS), and attenuated fibroblasts. FGFR-1 immunoreactivity was downregulated, and syndecan-4 immunoreactivity was upregulated in the basal cells. This suggests that FGF-2 in basal cells and LIS may be bound to the syndecan-4. We conclude that ozone and HDMA plus ozone effected incorporation of perlecan into the BMZ, resulting in atypical development of the BMZ. These changes are associated with specific alterations in the regulation of FGF-2, FGFR-1, and syndecan-4 in the airway epithelial-mesenchymal trophic unit, which may be associated with the developmental problems of lungs associated with exposure to ozone.

Effects of sex and hormonal status on astrocytic basic fibroblast growth factor-2 and tyrosine hydroxylase immunoreactivity after medial forebrain bundle 6-hydroxydopamine lesions of the midbrain dopamine neurons

We examined astrocytic basic fibroblast growth factor immunoreactivity (FGF-2-IR) and tyrosine hydroxylase immunoreactivity (TH-IR) in the cell-body region of midbrain dopaminergic neurons after unilateral infusions of the neurotoxin 6-hydroxydopamine into the medial forebrain bundle in male and female rats. In addition, to determine whether neonatal exposure to gonadal hormones has consequences on the expression of astrocytic FGF-2 and cell loss in response to injury in adulthood, we studied the effects of these lesions in adult male and female rats that had been exposed or not to testosterone in the neonatal period. In both males and females there was a progressive loss of TH-expressing cells that peaked 5 weeks after the lesions. Females showed less loss of TH-expressing cells than males, but this effect was not estrogen dependent. Lesions led to an increase in expression of astrocytic FGF-2 that was greater in males than in females. Finally, it was found that, regardless of genetic sex, rats exposed to testosterone neonatally showed greater astrocytic FGF-2 expression after lesions than those not exposed, and that among those not exposed to testosterone, estrogen treatment had a modest protective effect. Analysis of behavior and striatal dopamine content showed that the percent of striatal dopamine depletion 14 days after the lesion correlated with the amount of behavioral asymmetry displayed by animals on all tests conducted after lesioning. In groups killed 2 and 5 weeks after the lesion, the amount of behavioral asymmetry correlated with the percent loss of TH-IR cells and with the percent increase in FGF-2-IR cells in the midbrain. These relationships were not evident in groups killed 3 and 7 days after the lesion, possibly because the changes in the number of FGF-2- and TH-IR cells were not fully manifested. The present findings show that hormonal events early in life can alter the response of midbrain dopamine neurons to insult and injury in adult life and suggest that the slow degeneration of these neurons may release signals triggering a sustained activation of adjacent astrocytes which, in turn, may lead to induction of astrocytic FGF-2.

Fibroblast Growth Factor-2 in Remodeling of the Developing Basement Membrane Zone in the Trachea of Infant Rhesus Monkeys Sensitized and Challenged with Allergen

Remodeling of the epithelial basement membrane zone (BMZ) involves increased deposition of collagen, resulting in thickening of the BMZ. The current study focuses on fibroblast growth factor-2 (FGF-2) in the tracheal BMZ in house dust mite allergen (HDMA)-sensitized infant rhesus monkeys, challenged with HDMA at a time when the BMZ is undergoing active postnatal development. To detect structural changes in the BMZ, we measured collagens I, III, and V. To detect changes in the function of the BMZ, we measured immunoreactivity of the heparan sulfate proteoglycan, perlecan, and FGF-2. We found significant thickening of the tracheal BMZ (p < 0.05) with each of these parameters. We also found that all HDMA tracheal samples expressed thin focal areas of the BMZ associated with leukocyte trafficking. These areas were depleted of perlecan and FGF-2; however, increased FGF-2 immunoreactivity was present in the adjacent basal cells. We conclude that basal cells and FGF-2 are involved with significant remodeling of the BMZ in the developing trachea of infant rhesus monkeys exposed to HDMA.

Propentofylline after focal cortical lesion in the rat: impact on functional recovery and basic fibroblast growth factor expression

The effects of propentofylline, a xanthine derivative and adenosine transport inhibitor, were evaluated following anteromedial cortex lesion in the rat. Propentofylline (2×/10 mg/kg, intraperitoneally) was administered for 7 days post-insult and basic fibroblast growth factor (bFGF) immunoreactivity measured at designated time points in the peri-lesional cortex and ipsilateral dorsal striatum. The spatiotemporal pattern of bFGF expression was then compared to functional recovery patterns. Propentofylline-treated animals displayed increased bFGF expression in the peri-lesional cortex which may have contributed to the observed early facilitation of functional recovery. Drug administration did not, however, produce a change in bFGF expression in the ipsilateral dorsal striatum compared to saline-treated animals. These findings taken together with other positive findings regarding propentofylline, support the drug's therapeutic potential.