Nerve Growth Factor Immunoreactivity Research Articles

Morphometric, immunohistochemical and ultrastructural examination of age-related structural alterations in optic nerve

Aims: As individuals age, there is a known decline in visual function attributed to a reduction in the optic nerve fibers and myelin sheath degeneration. Studies present conflicting findings on whether aging affects axonal integrity in the human optic nerve. This study aims to investigate degenerative changes in the aging rat optic nerve. Methods: The investigation involved 36 Wistar albino rats. The rats were divided into six groups: the newborn, prepubertal, pubertal, junior, adult, and elderly groups. This study investigated optic nerve axon counts, axon diameters, levels of glial fibrillary acidic protein immunoreactivity (GFAP-IR) and nerve growth factor immunoreactivity (NGF-IR), as well as findings from light microscopy (LM) and electron microscopy (EM) in these groups. Results: This study observed age-related alterations in rat optic nerves, including increased diameter, irregular axon count fluctuations (both increases and decreases), elevated astrocyte count, and a simultaneous decline in oligodendrocyte count. Additionally, it was observed that NGF-IR was predominantly at the membrane level in newborns and moderately in the cytoplasm, whereas in older ages, it was evident at both cellular and axonal levels furthermore, it was observed that GFAP-IR increased with age. However, in LM and EM examinations, axonal loss and rarefaction, accumulation of osmiophilic substances, splitting of the myelin sheath, vacuolization, axonal retraction were observed. Conclusion: In this study, it was found that one of the causes of age-related vision loss is the advanced degenerative changes in the optic nerve and it was concluded that the remaining small-diameter myelinated nerve fibers may partially compensate for the sense of vision. Our study reveals that age-related degenerative changes in the central nervous system resemble those in multiple sclerosis (MS), suggesting a potential contribution to MS pathogenesis.

Seasonal variation of NGF in seminal plasma and expression of NGF and its cognate receptors NTRK1 and p75NTR in the sex organs of rams

Nerve growth factor (NGF) has long been known as the main ovulation-inducing factor in induced ovulation species, however, recent studies suggested the NGF role also in those with spontaneous ovulation. The first aim of this study was to evaluate the presence and gene expression of NGF and its cognate receptors, high-affinity neurotrophic tyrosine kinase 1 receptor (NTRK1) and low-affinity p75 nerve growth factor receptor (p75NTR), in the ram genital tract. Moreover, the annual trend of NGF seminal plasma values was investigated to evaluate the possible relationship between the NGF production variations and the ram reproductive seasonality. The presence and expression of the NGF/receptors system was evaluated in the testis, epididymis, vas deferens ampullae, seminal vesicles, prostate, and bulbourethral glands through immunohistochemistry and real-time PCR (qPCR), respectively. Genital tract samples were collected from 5 adult rams, regularly slaughtered at a local abattoir. Semen was collected during the whole year weekly, from 5 different adult rams, reared in a breeding facility, with an artificial vagina. NGF seminal plasma values were assessed through the ELISA method. NGF, NTRK1 and p75NTR immunoreactivity was detected in all male organs examined. NGF-positive immunostaining was observed in the spermatozoa of the germinal epithelium, in the epididymis and the cells of the secretory epithelium of annexed glands, NTRK1 receptor showed a localization pattern like that of NGF, whereas p75NTR immunopositivity was localized in the nerve fibers and ganglia. NGF gene transcript was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.01) in the testis than in the other tissues. NTRK1 gene transcript was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.05) in all the other tissues examined. Gene expression of p75NTR was highest (p < 0.01) in the seminal vesicles and lowest (p < 0.01) in the testis and bulbourethral glands. NGF seminal plasma concentration was greater from January to May (p < 0.01) than in the other months. This study highlighted that the NGF system was expressed in the tissues of all the different genital tracts examined, confirming the role of NGF in ram reproduction. Sheep are short-day breeders, with an anestrus that corresponds to the highest seminal plasma NGF levels, thus suggesting the intriguing idea that this factor could participate in an inhibitory mechanism of male reproductive activity, activated during the female anestrus.

Histopathological, immunohistochemical and biochemical evaluation of electroacupuncture treatment of nervus radialis and nervus ulnaris injuries in rabbits.

The aim of this study was to evaluate the efficacy of electroacupuncture in acute and chronic phases of radial and ulnar nerve injuries in histopathological, immunohistochemical and biochemical aspects. In the study, the rabbits were divided into four groups namely acute nerve injury (ANI) group, chronic nerve injury (CNI) group, positive control (PC) group and negative control (NC) group. In the ANI, CNI and PC groups, damage was created on the nervus radialis and nervus ulnaris by applying pressure for 60 seconds using a hemostatic forceps under anesthesia. No damage was created in the NC group. Fifteen sessions of electroacupuncture were applied to the rabbits in the ANI, CNI, and NC groups every other day using LI-4 (Large Intestine Meridian-4, He Gu), LI-10 (Large Intestine Meridian-10, Shou San Li), LR-3 (Liver Meridian-3, Tai Chong), and ST-36 (Stomach Meridian-36, Zusanli) electroacupuncture points. Electroacupuncture was not applied to the rabbits in the PC group. Decapitation was performed under general anesthesia at the end of electroacupuncture applications. After the euthanasia procedure, the samples obtained were evaluated for histopathological, immunohistochemical and biochemical parameters. In conclusion, degenerative foci in the treatment groups were found to be fewer than in the PC group whereas NGF and S-100 immunoreactivity were higher in the treatment groups than in the PC group. Whereas no statistically significant difference was observed between the treatment groups and the NC group in terms of oxidative stress factors, there was a statistically significant difference between the treatment groups and the PC group. In light of all these data, we have concluded that electroacupuncture is an effective treatment method for peripheral nerve injuries.

Probiotic supplementation alleviates absence seizures and anxiety- and depression-like behavior in WAG/Rij rat by increasing neurotrophic factors and decreasing proinflammatory cytokines

AimEpilepsy is one of the most common chronic brain disorders that affect millions of people worldwide. In the present study, we investigated the effects of probiotic supplementation on absence epilepsy and anxiety-and depression-like behavior in WAG/Rij rats. Material and methodFourteen male WAG/Rij rats (absence-epileptic) and seven male Wistar rats (nonepileptic) were used. The effects of probiotic VSL#3 (12.86 bn living bacteria/kg/day for 30 day/gavage) on absence seizures, and related psychiatric comorbidities were evaluated in WAG/Rij rats. Anxiety-like behavior was evaluated by the open-field test and depression-like behavior by the forced swimming test. In addition, the brain tissues of rats were evaluated histopathologically for nerve growth factor [NGF], brain-derived neurotrophic factor [BDNF], SRY sex-determining region Y-box 2 [SOX2] and biochemically for nitric oxide [NO], tumor necrosis factor-alpha [TNF-α] ,and Interleukin-6 [IL-6]. ResultsCompared to Wistar rats, WAG/Rij rats exhibited anxiety- and depression-like behavior, and had lower BDNF, NGF and SOX2 immunoreactivity, and higher TNF-α, IL-6 levels in brain tissue. VSL#3 supplementation reduced the duration and number of spike-wave discharges (SWDs) and exhibited anxiolytic or anti-depressive effect. VSL#3 supplement also increased the NGF immunoreactivity while decreasing IL-6, TNF-α and NO levels in WAG/Rij rat brain. ConclusionThe findings of the present study showed that neurotrophins, SOX2 deficiency, and pro-inflammatory cytokines may play a role in the pathogenesis of absence epilepsy. Our data support the hypothesis that the probiotics have anti-inflammatory effect. The present study is the first to show the positive effects of probiotic bacteria on absence seizures and anxiety- and depression-like behavior.

An immunohistochemical examination of cinnamon extract administration on distribution of NGF (nerve growth factor) and Trk-A (tyrosine kinase-A) receptor for diabetic rats with pancreatic tissue.

Background/aim The aim of this study was to investigate the administration of cinnamon extract that is known to be effective in decreasing the high blood glucose and the distribution of NGF and Trk-A receptor in pancreas with immunohistochemistry way.Materials and methods The experimental groups were defined as control, sham, cinnamon, diabetes, and diabetes-cinnamon. At the end of the experiment, the pancreatic tissue samples were obtained for the rats. The hematoxylin-eosin and triple staining were used to examine histology. The immunohistochemical methods were performed on the sections of pancreatic tissue. In all groups, the body weight and fasting blood glucose obtained from the male and female rats and the values were statistically evaluated.Results The NGF immunoreactivity was observed in acinus, excretory pars, excretorius ducts, and islets of Langerhans for the pancreatic tissues of female and male rats in all groups. The Trk-A immunoreactivity was observed in acinus and islets of Langerhans for the pancreatic tissues of female and male rats in the control, sham, and cinnamon groups.Conclusion As a result, it was determined that the cinnamon, which is effective on blood glucose levels, has a positive effect on the NGF production in pancreas.

The TrkA Antagonist AR786 Prevents Heterotopic Ossification

Soft tissue trauma can result in the pathologic formation of cartilage and bone outside of the skeleton, termed heterotopic ossification (HO). Therapeutic options for treatment of HO are lacking. We recently recognized the role of sensory nerve fibers in trauma-induced HO through nerve growth factor (NGF) and tropomyosin receptor kinase A (TrkA) signaling whereby the deletion of NGF-TrkA signaling impairs axonal invasion and blunts osteochondral differentiation after HO induction in a mouse model. Given this observation, we hypothesized that utilization of a small molecule antagonist for TrkA in clinical development, AR786, would demonstrate therapeutic benefit in the prevention of HO. Association of NGF and innervation with human HO of the posterior longitudinal ligament (HOPLL) was analyzed by microarray and immunohistochemistry. In a mouse model, trauma-induced HO was generated by the combination of cutaneous burn and Achilles tenotomy in 10 week old C57BL/6J mice, which causes HO over a well characterized 9 week time period. NGF expression was examined by single cell sequencing, immunohistochemistry, and western blot. Dorsal root ganglion (DRG) from the innervating lumbar tracts were examined for pTrkA expression by immunohistochemistry. The phenotypic consequences of TrkA inhibition by AR786 on HO formation was next assessed (Vehicle, n=5 and AR786, n=5). Nerve innervation within the injury site was visualized using immunohistochemistry stained with Beta III Tubulin (TUBB3) and confocal microscopy. Abnormal chondrogenesis was assessed by cartilage histomorphometry, and immunohistochemical detection (SOX9 and COL2). Data were presented as Means ± SD. Results showed that NGF and associated downstream NGF signaling were increased among HOPLL patient cells. NGF immunoreactivity was present in perivascular cells adjacent to HO, fibroblastic cells, and bone-associated cells. Increased nerve fibers were present surrounding HO and axonal sprouting was confirmed by neural antigen S100 and neurofilament SM31 in human HO tissue sections. During HO progression in mice, NGF, proNGF, and pTrkA were upregulated following HO induction. Nerve frequency was significantly reduced with AR786 treated animals compared to vehicle control in uninjured (p<0.001, Vehicle; 548.2 ± 118.0 vs. AR786; 104.2 ± 47.9, µm3) and 3-weeks post-injury (p<0.001, Vehicle; 3255.6 ± 1405.1 vs. AR786; 462.8 ± 201.1, µm3). Consequently, reduced axonal ingrowth significantly delays in ectopic cartilage formation (p<0.01, Vehicle; 0.33 ± 0.07 vs. AR786; 0.07 ± 0.06, mm2). In conclusion, NGF-TrkA signaling mediates crucial functions in human and rodent HO, and inhibition of TrkA-expressing neural inputs by AR786 abrogates HO progression. These findings suggest that NGF-TrkA signaling inhibition can be a potential therapeutic use in HO patients as negative regulator of HO disease progression.

The effect of erythropoietin and umbilical cord-derived mesenchymal stem cells on nerve regeneration in rats with sciatic nerve injury

ObjectiveWe aimed to investigate the effects of umbilical cord-derived mesenchymal stem cells and erythropoietin on nerve regeneration in the sciatic nerve ‘crush injury’ in a rat model. MethodsExperimental animals were randomly divided into 5 groups: Crush Injury, Sham, Crush Injury + Erythropoietin, Crush Injury + Mesenchymal Stem Cell, Crush Injury + Erythropoietin + Mesenchymal Stem Cell groups. Crush injury made with bulldog clamp. Mesencyhmal stem cells delivered by enjection locally. Erythropoietin administered by intraperitoneally. On the 0th, 14th and 28th days, all groups underwent a sciatic functional index test. On 28th day, sciatic nerves were harvested and histopathological appearance, axon number and axon diameter of the sciatic nerves were evaluated with Oil Red O staining. Immunoreactivity of nerve growth factor, neurofilament-H and caspase-3 were determined by immunofluorescence staining in nerve tissue. ResultsIn histopathological examination, axons and nerve bundles exhibiting normal nerve architecture in the Sham group. Crush Injury + Mesenchymal Stem Cell group has similar histological appearance to the Sham group. The number of axons were higher in the Mesenchymal Stem Cell groups compared to the Crush Injury group. Nerve growth factor immunoreactivity intensity was significantly lower in Crush Injury + Mesenchymal Stem Cell group compared to Crush Injury group. Neurofilament-H density was higher in the treatment groups when compared to the Crush Injury group. ConclusionsIn this study, it was found that umbilical cord-derived mesenchymal stem cells and erythropoietin treatments effects positively regeneration of crush injury caused by bulldog clamp in the sciatic nerve of rats.

The Distribution of Activating Transcription Factor 6 (ATF6) and Nerve Growth Factor (NGF) in the Duodenum Tissue of Diabetic and Non-Diabetic Rats

Objective: This study was conducted with the purpose of investigating the distribution of the Activating Transcription Factor 6 (ATF6) and the Nerve Growth Factor (NGF) in the duodenum tissue of diabetic and non-diabetic rats. Material and Method: Eighteen female Sprague Dawley rats were randomly divided into three groups as the control, sham, and diabetes groups. Routine histological and immunohistochemical methods were applied to the duodenum tissues collected at the end of the study. Results: It was determined that the villus length measurements showed a statistically significant difference between the control and diabetes groups. There was NGF immunoreactivity which was moderate and diffuse cytoplasmic in the villus intestinalis and muscularis layer in all groups, weak in the crypts and glands in the control and sham groups, and moderate and diffuse cytoplasmic in the diabetes group. ATF6 immunoreactivity was determined moderate in the villus intestinalis, crypts, glands, and muscularis layer in the control and sham groups and strong diffuse cytoplasmic in the diabetes group. Conclusion: It was determined that both NGF and ATF6 immunoreactivity increased in the duodenum tissue of the rats on which diabetes was induced experimentally.

Neuroanatomical basis of the nerve growth factor ovulation-induction pathway in llamas†.

The objective of the study was to characterize the anatomical framework and sites of action of the nerve growth factor (NGF)-mediated ovulation-inducing system of llamas. The expression patterns of NGF and its receptors in the hypothalamus of llamas (n=5) were examined using single and double immunohistochemistry/immunofluorescence. We also compare the expression pattern of the P75 receptor in the hypothalamus of llama and a spontaneous ovulator species (sheep, n=5). Both NGF receptors (TrkA and P75) were highly expressed in the medial septum and diagonal band of Broca, and populations of TrkA cells were observed in the periventricular and dorsal hypothalamus. Unexpectedly, we found NGF immunoreactive cell bodies with widespread distribution in the hypothalamus but not in areas endowed with NGF receptors. The organum vasculosum of the lamina terminalis (OVLT) and the median eminence displayed immunoreactivity for P75. Double immunofluorescence using vimentin, a marker of tanycytes, confirmed that tanycytes were immunoreactive to P75 in the median eminence and in the OVLT. Additionally, tanycytes were in close association with GnRH and kisspeptin in the arcuate nucleus and median eminence of llamas. The choroid plexus of llamas contained TrkA and NGF immunoreactivity but no P75 immunoreactivity. Results of the present study demonstrate sites of action of NGF in the llama hypothalamus, providing support for the hypothesis of a central effect of NGF in the ovulation-inducing mechanism in llamas.

Effect of electroacupuncture on glial fibrillary acidic protein and nerve growth factor in the hippocampus of rats with hyperlipidemia and middle cerebral artery thrombus.

Electroacupuncture (EA) has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia. However, there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects. In this study, EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed. Except for the normal group, rats in other groups were fed a high-fat diet throughout the whole experiment. Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks. Middle cerebral artery thrombus models were induced by pasting 50% FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group. EA1 group rats received EA at bilateral ST40 (Fenglong) for 7 days before the thrombosis. Rats in the EA1 and EA2 groups received EA at GV20 (Baihui) and bilateral ST40 for 14 days after model establishment. Neuronal health was assessed by hematoxylin-eosin staining in the brain. Hyperlipidemia was assessed by biochemical methods that measured total cholesterol, triglyceride, low-density lipoprotein and high-density lipoprotein in blood sera. Behavioral analysis was used to confirm the establishment of the model. Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region. The results demonstrated that, compared with the model group, blood lipid levels significantly decreased, glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups. The repair effect was superior in the EA1 group than in the EA2 group. These findings confirm that EA can reduce blood lipid, inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis. All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine, China (approval No. BUCM-3-2018022802-1002) on April 12, 2018.

Preliminary Evaluation of Seminal Plasma Proteins and Immunoreactivity of Nerve Growth Factor as Indicative of an Ovulation Inducing Factor in Odontocetes

In the seminal plasma of terrestrial mammalian species known as induced (e.g., camels) and spontaneous (e.g., cattle) ovulators, an ovulation-inducing factor (OIF) with a protein structure similar to beta-nerve growth factor (β-NGF) has been identified. Detection of an OIF/NGF in the seminal plasma of cetaceans would have both basic and applied implications in reproductive biology and conservation management programs. A preliminary evaluation was conducted to characterize the distribution and abundance of seminal plasma proteins in aquarium-based belugas and a Pacific white-sided and bottlenose dolphin. Initially, SDS-PAGE was used with 50 μg of total protein for separation; thereafter, Western immunoblot was used with anti-NGF. In addition to odontocete seminal plasma, a purified fraction of llama seminal plasma (100 ng protein) and an extract of mouse brain (20 μg total protein) were included as positive controls for NGF. Within the two belugas, visual inspection of the protein bands indicated similar distribution and intensity. However, among the belugas and Pacific white-sided and bottlenose dolphins there was more diversity than similarity in the distribution and abundance of seminal plasma proteins. While immunoreactivity of NGF was distinctly evident in the llama and mouse positive controls, there was no visual reactivity in any of the odontocete samples. These preliminary results provide novel information indicating more homogeneity within and heterogeneity among seminal plasma proteins of ondentocetes. Although NGF was not immunologically detected, future studies are required to address the apparent limitations of immuno-detection of NGF, especially if the post-translational form of β-NGF is in low abundance in the seminal plasma of belugas and Pacific white-sided and bottlenose dolphins.

Associations of Symptomatic Knee Osteoarthritis With Histopathologic Features in Subchondral Bone.

Subchondral bone and the osteochondral junction are thought to contribute to osteoarthritis (OA) knee pain. We undertook this study to identify osteochondral pathologies specifically associated with symptomatic human knee OA. Medial tibial plateau samples from 2 groups of subjects (n = 31 per group) were matched for macroscopic chondropathy scores. The symptomatic chondropathy group had undergone total knee replacement for OA knee pain, at which time specimens of the medial tibial plateau were obtained. The asymptomatic chondropathy group included subjects who died of unrelated illness (specimens were obtained at postmortem examination) and who had not previously sought help for knee pain. OA histopathology, immunoreactivity for nerve growth factor (NGF) and CD68 (macrophages), tartrate-resistant acid phosphatase-positive subchondral osteoclasts, and synovitis were compared between groups. Mankin scores, subchondral bone density, and subchondral CD68-immunoreactive macrophage infiltration were similar between the 2 groups. NGF-like immunoreactivity was found in subchondral mononuclear cells and osteoclasts, as well as in chondrocytes. NGF in osteochondral channels and osteoclast densities in subchondral bone were higher in the symptomatic chondropathy group than in the asymptomatic chondropathy group (P < 0.01 and P = 0.02, respectively), as were synovitis scores (P < 0.01). Osteochondral pathology was not significantly associated with synovitis score. The differences in NGF expression and in osteoclast density remained significant after adjustment for age and synovitis score (P = 0.01 and P = 0.04, respectively). Osteochondral NGF and osteoclast densities, together with synovitis scores, explained ~28% of sample allocation to symptomatic or asymptomatic groups. Subchondral pathology was associated with symptomatic knee OA, independent of chondropathy and synovitis. Increased NGF expression in osteochondral channels and increased osteoclast density appear to be key features associated with bone pain in knee OA.

MP45-06 BLADDER UROTHELIAL AND AFFERENT DYSFUNCTIONS UNDERLYING BLADDER OVERACTIVITY IN RATS WITH CHEMICALLY INDUCED PROSTATIC INFLAMMATION

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2018MP45-06 BLADDER UROTHELIAL AND AFFERENT DYSFUNCTIONS UNDERLYING BLADDER OVERACTIVITY IN RATS WITH CHEMICALLY INDUCED PROSTATIC INFLAMMATION Shinsuke Mizoguchi, Amanda S. Wolf-Johnson, Takahisa Suzuki, Eiichiro Takaoka, Donald B. DeFranco, Zhou Wang, Lori A. Birder, and Naoki Yoshimura Shinsuke MizoguchiShinsuke Mizoguchi More articles by this author , Amanda S. Wolf-JohnsonAmanda S. Wolf-Johnson More articles by this author , Takahisa SuzukiTakahisa Suzuki More articles by this author , Eiichiro TakaokaEiichiro Takaoka More articles by this author , Donald B. DeFrancoDonald B. DeFranco More articles by this author , Zhou WangZhou Wang More articles by this author , Lori A. BirderLori A. Birder More articles by this author , and Naoki YoshimuraNaoki Yoshimura More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1445AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES There is increasing evidence showing the positive correlation between prostatic inflammation and lower urinary tract symptoms (LUTS) in males with benign prostatic hyperplasia (BPH). Therefore, this study used a rat model of non-bacterial prostatic inflammation to investigate the changes in bladder activity and molecular characteristics of bladder afferent pathways and urothelium after prostatic inflammation. METHODS Male SD rats were used, and prostatic inflammation was induced by formalin (5%; 50 µl per lobe) injection into bilateral ventral lobes of the prostate. Bladder function was evaluated 1 and 4 weeks after formalin injection using awake cystometry. Also, after 10 days of prostatic inflammation, prostaglandin E2 (PGE2) concentrations in bladder tissues were measured by ELISA. In bladder mucosa, protein levels of TRPV1 and PGE2 receptors (EP1 to 4) as well as mRNA levels of ASICs (1-3), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were evaluated by Western blot and RT-PCR, respectively. NGF immunoreactivity in bladder sections was also examined. In addition, mRNA levels of TRPV1 and TRPA1 receptors were measured in bladder afferent neurons labeled by Fast Blue (FB) injected into the bladder wall using laser-capture microdissection and RT-PCR methods. RESULTS Compared to vehicle-injected rats, formalin-treated rats exhibited a significant (p<0.05) decrease in intercontraction intervals in cystometry at 1 and 4 weeks after formalin injection. Tissue inflammation evident as inflammatory cell infiltration was found in the prostate, but not in the bladder from formalin-injected rats. In rats with prostatic inflammation (10 days), bladder PGE2 concentrations were significantly increased, and protein levels of TRPV1 and EP4 receptors as well as mRNA levels of all ASICs, BDNF and NGF were significantly elevated in bladder mucosa of formalin-injected rats. NGF immunoreactivity was also increased in bladder urothelium after prostatic inflammation. In FB-labeled bladder afferent neurons, mRNA levels of TRPV1 and TRPA1 were increased compared to non-labelled neurons in prostatic inflammation rats. CONCLUSIONS These results indicate that non-bacterial inflammation confined in the prostate induces bladder overactivity, enhanced PGE2 production in the bladder and upregulations of receptors and growth factors in bladder urothelium and/or afferent pathways, which could contribute to storage LUTS in BPH patients with prostatic inflammation. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e599 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Shinsuke Mizoguchi More articles by this author Amanda S. Wolf-Johnson More articles by this author Takahisa Suzuki More articles by this author Eiichiro Takaoka More articles by this author Donald B. DeFranco More articles by this author Zhou Wang More articles by this author Lori A. Birder More articles by this author Naoki Yoshimura More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Nerve Growth Factor Is Associated With Sexual Pain in Women With Endometriosis.

Endometriosis is present in 1 in 10 reproductive-age women, and half experience deep dyspareunia (pelvic pain with sexual intercourse). Our objective was to investigate nerve growth factor (NGF) and its receptors (TrkA/p75NTR) in endometriosis-associated deep dyspareunia. A total of 32 women with endometriosis in the posterior pelvic compartment (cul-de-sac/uterosacrals) were included, either with (n = 17) or without (n = 15) deep dyspareunia symptoms confirmed by endovaginal ultrasound-assisted palpation on examination. Expression of NGF, TrkA, and p75NTR in the surgically excised cul-de-sac/uterosacral endometriosis was examined by immunohistochemistry and Histoscore blinded to pain phenotypes. Additionally, endometriotic stromal cells (ESCs; n = 3) from ectopic endometriosis lesions were cultured and incubated with/without NGF and/or Trk inhibitor K252a, followed by expression analysis of prostaglandin-endoperoxide synthase 2 (PTGS-2)/cyclooxygenase 2 (COX-2; reverse transcription quantitative real-time polymerase chain reaction and Western blot) and prostaglandin E2 (PGE2) secretion (enzyme-linked immunosorbent assay). We found that the immunointensity of NGF and TrkA, but not p75NTR, was significantly elevated in endometriotic stroma and epithelium from women with deep dyspareunia compared to women without deep dyspareunia. Nerve growth factor immunoreactivity in the stroma was also significantly associated with deep dyspareunia intensity and local nerve bundle density. In cultured ESCs, NGF significantly increased PTGS-2/COX-2 mRNA and protein levels as well as PGE2 secretion, and these effects could be abolished by pretreatment of Trk inhibitor K252a. In conclusion, elevated NGF levels were associated with deep dyspareunia in women with cul-de-sac/uterosacral endometriosis. This association may be mediated by increased nerve bundle density and by COX-2/PGE2 stimulation via Trk receptor. Nerve growth factor signaling may play an important role in endometriosis-associated sexual pain.

Evaluation of the immunoreactivity of nerve growth factor and tropomyosin receptor kinase A in the esophagus of noninfected and infected individuals with Trypanosoma cruzi

Megaesophagus is one of the major manifestations of the chronic phase of Chagas disease. Its primary symptom is generally dysphagia due to disturbance in the lower esophageal sphincter. Microscopically, the affected organ presents denervation, which has been considered as consequence of an inflammatory process that begins at the acute phase and persists in the chronic phase. Inflammatory infiltrates are composed of lymphocytes, macrophages, natural killer cells, mast cells, and eosinophils. In this study, we evaluated the immunoreactivity of nerve growth factor (NGF), and of its receptor tropomyosin receptor kinase A (TrkA), molecules that are well known for having a relevant role in neuroimmune communication in the gastrointestinal tract. Esophageal samples obtained via autopsy or surgery procedures from six noninfected individuals, six infected individuals without megaesophagus, and six infected individuals with megaesophagus were analyzed. Infected individuals without megaesophagus presented increased numbers of NGF immunoreactive (IR) mast cells and increased areas of TrkA-IR epithelial cells and inner muscle cells. Infected individuals with megaesophagus showed increased numbers of NGF-IR eosinophils and mast cells, TrkA-IR eosinophils and mast cells, increased area of NGF-IR epithelial cells, and increased areas of TrkA-IR epithelials cells and inner muscle cells. The data presented here point to the participation of NGF and its TrkA receptor in the pathology of chagasic megaesophagus.

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.

Constitutive expression Of NGF And P75NTR affected by bladder distension and NGF antisense treatment

AimsIt is known that bladder exposure to noxious stimuli elicits nerve growth factor (NGF) expression with region wise differences. Here, we investigated the effect of bladder distension (cystometry) and bladder wall injection of NGF antisense oligonucleotide (ODN) together as well as separately on spontaneous (constitutive) expression of NGF and its cognate p75 neurotrophin receptor (p75NTR). MethodUnder isoflurane anesthesia, either 15μg of protamine sulfate (vehicle) alone or complexed with 1.5μg of NGF antisense or scrambled ODN was injected (10μL) at 4 sites in bladder wall of 24 adult female Sprague-Dawley rats and 6 rats were left untreated (n=30). Under urethane anesthesia, cystometry (CMG) was performed in treated and control rats. Fluorescent ODN and NGF/p75NTR expression was localized in harvested tissue. Key findingsComplexation of ODN with protamine was essential for the retention of ODN in bladder tissue as the uncomplexed ODN was untraceable after injection. Bladder distension from CMG raised the expression of NGF and p75NTR relative to CMG naïve rats. The groups treated with vehicle, scrambled and antisense ODN were indistinct with regard to CMG parameters, but the intense immunoreactivity of NGF and p75NTR seen in the vehicle and scrambled ODN groups was reduced following treatment with NGF antisense. SignificanceThe constitutive expression of NGF and p75NTR is responsive to bladder distension and administration of NGF antisense. Complexation with protamine reduces the clearance of ODN and demonstrates the potential of ODN nanoparticles as an option for reducing the inducible NGF expression in OAB patients following intradetrusor injection.

Age-Related (Aged vs. Adult) Comparison of the Effect of Two Mild Stressors on the Nerve Growth Factor (NGF) in the Rat Hypothalamic Supraoptic Nucleus (SON) – Immunohistochemical Study

The ontogenetic period of life and stress can have different effects on the nerve growth factor (NGF) in the hypothalamus. The aim of our study was to investigate the influence of two mild stressors, acute and chronic exposure to forced swim (FS) or high-light open field (HL-OF), on neurons containing NGF. Immunofluorescence staining was used to reveal the density of NGF-immunoreactive (ir) cells in the hypothalamic supraoptic nucleus (SON) in adult (postnatal day 90; P90) and aged (P720) rats. The P90 and P720 rats that were subjected to acute and chronic FS showed no differences in the density of NGF-ir neurons in the SON compared with nonstressed rats. However, a significant increase in NGF-ir cells was noted after acute but not after chronic HL-OF only in P90 rats. What is more, there were no age-related (P90 vs. P720) changes in the density of NGF-ir neurons in non-stressed and FS- or HL-OF-stressed rats. Our results indicate that acute HL-OF was the only factor inducing changes in the density of NGF-ir neurons in the SON of adult rats. This could be related to the neuroprotective role of NGF-ir cells in response to acute HL-OF. The absence of age-dependent changes in the density of NGF-ir neurons may indicate that the ageing processes in SON do not generate changes in the NGF immunoreactivity of its neurons.

The impact of two mild stressors on the nerve growth factor (NGF) immunoreactivity in the amygdala in aged rats compared to adult ones

Nerve growth factor (NGF) seems to play an important role in the ageing limbic system in response to stress. This study aimed to explore the influence of acute and chronic exposure to high-light open field (HL-OF) or forced swim (FS) stressors on the density of NGF immunoreactive (ir) neurons in the amygdala central (CeA), medial (MeA), lateral (LA) and basolateral (BLA) nuclei in adult (postnatal day 90; P90) and aged (P720) rats. In comparison with non-stressed rats, neither acute nor chronic HL-OF produced significant changes in the density of NGF-ir neurons of studied nuclei in P90 and P720 rats. However, not acute but chronic FS was the factor inducing an increase in the density of NGF-ir neurons in the CeA of both age groups and in the LA of P720 rats. Despite the lack of change in the density of NGF-ir neurons between P90 and P720 non-stressed rats, there were significant age-related changes in NGF-ir cells in FS and/or HL-OF stressed rats in all the tested nuclei, with the exception of the LA. It may be concluded that as far as the influence on NGF-ir cells in amygdaloid nuclei is concerned, HL-OF did not constitute an aggravating factor for rats in the ontogenetic periods studied. Moreover, upregulation of NGF-ir neurons predominantly in CeA after chronic FS seems to be neuroprotective. Age-dependent changes in the density of NGF-ir neurons in stressed rats are probably caused by ageing processes and they may point to dysregulation of excitatory control exerted by the amygdala.

Expression of nerve growth factor and its receptor, tyrosine kinase receptor A, in rooster testes

Nerve growth factor (NGF), which is required for the survival and differentiation of the nervous system, is also thought to play an important role in the development of mammalian reproductive tissues. To explore the function of NGF in the male reproductive system of non-mammalian animals, we determined the presence of NGF and its receptor, tyrosine kinase receptor A (TrkA), in rooster testes and investigated the regulation of NGF and TrkA expression by follicle-stimulating hormone (FSH). The mRNA and protein levels of NGF and TrkA in 6-week-old rooster testes were lower than those in 12-, 16- or 20-week age groups; levels were highest in the 16-week group. Immunohistochemistry showed that NGF and TrkA were both detected in spermatogonia, spermatocytes and spermatids. NGF immunoreactivity was observed in Leydig cells and strong TrkA signals were present in Sertoli cells. Meanwhile, FSH increased TrkA transcript levels in rooster testes in a dose-dependent manner. We present novel evidence for the developmental and FSH-regulated expression of the NGF/TrkA system, and our findings suggest that the NGF/TrkA system may play a prominent role in chicken spermatogenesis.