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1.
Aquat Toxicol ; 223: 105482, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32371337

RESUMO

The use of online remote control for 24/7 behavioural monitoring can play a key role in estimating the environmental status of aquatic ecosystems. Recording the valve activity of bivalve molluscs is a relevant approach in this context. However, a clear understanding of the underlying disturbances associated with behaviour is a key step. In this work, we studied freshwater Asian clams after exposure to crude oil (measured concentration, 167 ± 28 µg·L-1) for three days in a semi-natural environment using outdoor artificial streams. Three complementary approaches to assess and explore disturbances were used: behaviour by high frequency non-invasive (HFNI) valvometry, tissue contamination with polycyclic aromatic hydrocarbons (PAH), and proteomic analysis. Two tissues were targeted: the pool adductor muscles - retractor pedal muscle - cerebral and visceral ganglia, which is the effector of any valve movement and the gills, which are on the frontline during contamination. The behavioural response was marked by an increase in valve closure-duration, a decrease in valve opening-amplitude and an increase in valve agitation index during opening periods. There was no significant PAH accumulation in the muscle plus nervous ganglia pool, contrary to the situation in the gills, although the latter remained in the low range of data available in literature. Major proteomic changes included (i) a slowdown in metabolic and/or cellular processes in muscles plus ganglia pool associated with minor toxicological effect and (ii) an increase of metabolic and/or cellular processes in gills associated with a greater toxicological effect. The nature of the proteomic changes is discussed in terms of unequal PAH distribution and allows to propose a set of explanatory mechanisms to associate behaviour to underlying physiological changes following oil exposure. First, the first tissues facing contaminated water are the inhalant siphon, the mantle edge and the gills. The routine nervous activity in the visceral ganglia should be modified by nervous information originating from these tissues. Second, the nervous activity in the visceral ganglia could be modified by its own specific contamination. Third, a decrease in nervous activity of the cerebral ganglia close to the mouth, including some kind of narcosis, could contribute to a decrease in visceral ganglia activity via a decrease or blockage of the downward neuromodulation by the cerebro-visceral connective. This whole set of events can explain the decrease of metabolic activity in the adductor muscles, contribute to initiate the catch mechanism and then deeply modify the valve behaviour.


Assuntos
Comportamento Animal/efeitos dos fármacos , Corbicula/efeitos dos fármacos , Corbicula/metabolismo , Petróleo/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Proteoma/metabolismo , Poluentes Químicos da Água/toxicidade , Animais , Ecossistema , Água Doce/química , Gânglios/efeitos dos fármacos , Gânglios/metabolismo , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Músculos/efeitos dos fármacos , Músculos/metabolismo , Proteômica
2.
PLoS One ; 15(4): e0231353, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32271824

RESUMO

Prohormone convertases (PCs) are subtilisin-like proteases responsible for the intracellular processing of prohormones and proneuropeptides in vertebrates and invertebrates. The full-length PC2 cDNA sequence was cloned from pleuropedal ganglion of Haliotis discus hannai, consisted of 2254-bp with an open reading frame of 1989-bp and encoded a protein of 662 amino acid residues. The architecture of Hdh PC2 displayed key features of PCs, including a signal peptide, a pro-segment domain with sites for autocatalytic activation, a catalytic domain, and a pro-protein domain (P-domain). It shares the highest homology of its amino acid sequence with the PC2 from H. asinina and to lesser extent with that of Homo sapiens and Rana catesbeiana PC2. Sequence alignment analysis indicated that Hdh PC2 was highly conserved in the catalytic domain, including a catalytic triad of serine proteinases of the subtilisin family at positions Asp-195, His-236, and Ser-412. The cloned sequence contained a canonical integrin binding sequence, and four cysteine residues involved in the formation of an intramolecular disulfide link. Phylogenetic analysis revealed that the Hdh PC2 is robustly clustered with the Has PC2. Quantitative PCR assay demonstrated that the Hdh PC2 was predominantly expressed in the pleuropedal ganglion rather than in other examined tissues. Although PC2 mRNA was expressed throughout the gametogenetic cycle of male and female abalone, the expression level was significantly higher in the ripening stage of female abalone. Also, a significantly higher expression was observed in the pleuropedal ganglion and gonadal tissues at a higher effective accumulative temperature (1000°C). In situ hybridization revealed that the PC2 mRNA expressing neurosecretory cells were distributed in the cortex region of the pleuropedal ganglion. According to the results, it can be concluded that pleuropedal ganglion is the highest site of PC2 activity, and this enzyme might be involved in the abalone reproduction process.


Assuntos
Gastrópodes/enzimologia , Pró-Proteína Convertase 2/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , Feminino , Gânglios/metabolismo , Gânglios/patologia , Gônadas/metabolismo , Hibridização In Situ , Filogenia , Pró-Proteína Convertase 2/classificação , Pró-Proteína Convertase 2/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Temperatura
3.
Zoolog Sci ; 37(1): 50-60, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32068374

RESUMO

The labial palps of bivalves are thought to be involved in suspension feeding. However, the function of their muscular movements and neural regulation are still unclear. In semi-intact preparations of Mytilus, in which one valve was removed, suspended particles were removed from the labial palps following two kinds of compound movements: torsional and rotational. Both of these compound movements are therefore thought to function in rejection during feeding. These movements were observed in reduced preparations of isolated labial palps with intact cerebral ganglia, and were maintained even after removal of the cerebral ganglia, suggesting that they are generated by the peripheral neural network. Stimulation of the anterior pallial nerve elicited tetanic contraction of the labial palp, followed by secondary responses, including torsional movement. Secondary responses were dramatically reduced by a high concentration of divalent cations, in which polysynaptic pathways were inhibited. Hence, the cerebral ganglia may play an excitatory role within the peripheral neural network and the labial palp musculature via the anterior pallial nerve. Administration of serotonin induced repetitive muscular movements, whereas dopamine did not induce muscular movements. Serotonin-induced muscular movements were not elicited under a high concentration of divalent cation condition. In histochemical experiments, both the serotonergic and dopaminergic neural processes and cell body-like structures were widely observed inside the labial palp, the anterior pallial nerve, and the cerebral ganglia. Serotonin may thus contribute to activation of polysynaptic peripheral pathways, which are involved in regulating compound movements.


Assuntos
Comportamento Alimentar , Músculos/inervação , Mytilus/fisiologia , Animais , Dopamina/farmacologia , Gânglios , Movimento/efeitos dos fármacos , Músculos/efeitos dos fármacos , Serotonina/farmacologia
4.
Fish Shellfish Immunol ; 96: 126-137, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31760166

RESUMO

Spiroplasma eriocheiris, a novel pathogen of Chinese mitten crab Eriocheir sinensis tremor disease, has led into catastrophic economic losses in aquaculture. S. eriocheiris invaded the hemocytes in the early stage, then invaded nerve tissue and caused typically paroxysmal tremors of pereiopod in the late stage of infection. The purpose of this study was to detect the infection mechanism of hemocytes in the early stage and thoracic ganglion in the late stage of S. eriocheiris infection at the protein level. Hemocytes and thoracic ganglion were collected at 24 h and 10 d after injection (the crabs with typical paroxysmal tremors of the pereiopod), respectively. TMT was performed with isobaric markers, followed by liquid chromatography tandem mass spectrometry (LC-MS/MS). In hemocytes, 127 proteins were up-regulated and 85 proteins were down-regulated in 2747 quantified proteins. Many proteins and process including proPO system proteins, hemolymph coagulation system proteins and lectins were differently expressed in hemocytes and involved in the early immune process of E. sinensis against S. eriocheiris infection. Meanwhile, 545 significantly different expression proteins (292 down-regulated and 253 up-regulated protein including a number of immune-associated, nervous system development and signal transmission related proteins) were identified in thoracic ganglion in the late stage of S. eriocheiris infection. The qRT-PCR analysis results shown that the selected significantly changed proteins in hemocytes and thoracic ganglion were consistent with the TMT proteomics. This paper reported for the first time to study the responses of crab hemocyte and thoracic ganglion against the S. eriocheiris infection at different stages. These findings help us understand the infection mechanism of S. eriocheiris at different stage with the different tissue.


Assuntos
Proteínas de Artrópodes/imunologia , Braquiúros/imunologia , Hemócitos/imunologia , Proteoma/imunologia , Spiroplasma/fisiologia , Animais , Braquiúros/microbiologia , Gânglios/imunologia , Proteômica
5.
Parasitol Res ; 119(1): 317-319, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31782012

RESUMO

Muscle cells of a digenean fish blood fluke, Aporocotyle simplex, aggregate along the periphery of the cerebral ganglia. Solitary myocytons and sarcoplasmic processes with muscle fibres give rise to long, narrow lamellate projections, which are visible along the periphery and within ganglia. These ultrastructural observations suggest a switching of glial functions to muscle cells and represent additional evidence of the phylogenetic lability of glial cells in bilaterians.


Assuntos
Células Musculares/classificação , Neuroglia/classificação , Schistosomatidae/citologia , Animais , Doenças dos Peixes/parasitologia , Gânglios/citologia , Células Musculares/citologia , Células Musculares/ultraestrutura , Neuroglia/citologia , Neuroglia/ultraestrutura , Schistosomatidae/anatomia & histologia , Schistosomatidae/ultraestrutura , Infecções por Trematódeos/parasitologia , Infecções por Trematódeos/veterinária
6.
Elife ; 82019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31868164

RESUMO

Myoclonus dystonia (DYT11) is a movement disorder caused by loss-of-function mutations in SGCE and characterized by involuntary jerking and dystonia that frequently improve after drinking alcohol. Existing transgenic mouse models of DYT11 exhibit only mild motor symptoms, possibly due to rodent-specific developmental compensation mechanisms, which have limited the study of neural mechanisms underlying DYT11. To circumvent potential compensation, we used short hairpin RNA (shRNA) to acutely knock down Sgce in the adult mouse and found that this approach produced dystonia and repetitive, myoclonic-like, jerking movements in mice that improved after administration of ethanol. Acute knockdown of Sgce in the cerebellum, but not the basal ganglia, produced motor symptoms, likely due to aberrant cerebellar activity. The acute knockdown model described here reproduces the salient features of DYT11 and provides a platform to study the mechanisms underlying symptoms of the disorder, and to explore potential therapeutic options.


Assuntos
Distúrbios Distônicos/genética , Distúrbios Distônicos/mortalidade , Sarcoglicanas/genética , Sarcoglicanas/metabolismo , Animais , Córtex Cerebelar/metabolismo , Cerebelo/metabolismo , Cerebelo/patologia , Modelos Animais de Doenças , Distonia , Distúrbios Distônicos/induzido quimicamente , Distúrbios Distônicos/patologia , Etanol/efeitos adversos , Feminino , Gânglios/metabolismo , Predisposição Genética para Doença/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , RNA Interferente Pequeno
7.
Bull Exp Biol Med ; 168(1): 76-78, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31768775

RESUMO

Expression of neuronal NO synthase in the sympathetic cranial cervical ganglion and stellate ganglion in rats during postnatal ontogeny was studied by immunohistochemistry and Western blotting. In the sympathetic ganglia, neuronal NO synthase-immunoreactive neurons were absent in all rats. In the stellate and cranial cervical ganglia, the expression of neuronal NO synthase and the density of immunoreactive fibers increased in early postnatal ontogeny from the moment of birth to the age of 30 days and then decreased. Thus, we observed heterochroneous expression of neuronal NOS in the preganglionic somata in the spinal cord and in the preganglionic fibers in the sympathetic ganglia during ontogeny.


Assuntos
Gânglios Simpáticos/metabolismo , Gânglios/metabolismo , Óxido Nítrico Sintase/metabolismo , Sistema Nervoso Simpático/citologia , Animais , Western Blotting , Feminino , Imuno-Histoquímica , Ratos , Ratos Wistar
8.
Proc Natl Acad Sci U S A ; 116(45): 22833-22843, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31636217

RESUMO

Birdsong, like human speech, consists of a sequence of temporally precise movements acquired through vocal learning. The learning of such sequential vocalizations depends on the neural function of the motor cortex and basal ganglia. However, it is unknown how the connections between cortical and basal ganglia components contribute to vocal motor skill learning, as mammalian motor cortices serve multiple types of motor action and most experimentally tractable animals do not exhibit vocal learning. Here, we leveraged the zebra finch, a songbird, as an animal model to explore the function of the connectivity between cortex-like (HVC) and basal ganglia (area X), connected by HVC(X) projection neurons with temporally precise firing during singing. By specifically ablating HVC(X) neurons, juvenile zebra finches failed to copy tutored syllable acoustics and developed temporally unstable songs with less sequence consistency. In contrast, HVC(X)-ablated adults did not alter their learned song structure, but generated acoustic fluctuations and responded to auditory feedback disruption by the introduction of song deterioration, as did normal adults. These results indicate that the corticobasal ganglia input is important for learning the acoustic and temporal aspects of song structure, but not for generating vocal fluctuations that contribute to the maintenance of an already learned vocal pattern.


Assuntos
Comunicação Animal , Córtex Cerebral/fisiologia , Gânglios/fisiologia , Aprendizagem , Neurônios/fisiologia , Aves Canoras/fisiologia , Animais , Córtex Cerebral/citologia , Gânglios/citologia
9.
Pol J Vet Sci ; 22(2): 427-430, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31269360

RESUMO

Combined retrograde tracing and double-labelling immunofluorescence were used to investigate the distribution and chemical coding of neurons in aorticoerenal (ARG) and testicular (TG) ganglia supplying the urinary bladder apex (UBA) in the juvenile male pig (n=4, 12 kg. body weight). Retrograde fluorescent tracer Fast Blue (FB) was injected into the wall of the bladder apex under pentobarbital anesthesia. After three weeks all the pigs were deeply anesthetized and transcardially perfused with 4% buffered paraformaldehyde. TG and ARG were collected and processed for double-labelling immunofluorescence. The presence of tyrosine hydroxylase (TH) or dopamine beta-hydroxylase (DBH), neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL), nitric oxide synthase (NOS) and vesicular acetylcholine transporter (VAChT) were investigated. The cryostat sections were examined with a Zeiss LSM 710 confocal microscope equipped with adequate filter blocks. The TG and ARG were found to contain many FB-positive neurons projecting to the UBA (UBA-PN). The UBA-PN were distributed in both TG and ARG. The majority were found in the left ganglia, mostly in TG. Immunohistochemistry disclosed that the vast majority of UBA-PN were noradrenergic (TH- and/or DBH-positive). Many noradrenergic neurons also contained immunoreactivity to NPY, SOM or GAL. Most of the UBA-PN were supplied with varicose VAChT-, or NOS- IR (immunoreactive) nerve fibres. This study has revealed a relatively large population of differently coded ARG and TG neurons projecting to the porcine urinary bladder. As judged from their neurochemical organization these nerve cells constitute an important element of the complex neuro-endocrine system involved in the regulation of the porcine urogenital organ function.


Assuntos
Aorta/inervação , Gânglios/citologia , Rim/inervação , Suínos , Testículo/inervação , Bexiga Urinária/inervação , Animais , Gânglios/fisiologia , Masculino
10.
BMC Cancer ; 19(1): 633, 2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31242896

RESUMO

BACKGROUND: The recent implementation of PET with prostate specific membrane antigen (PSMA)-specific radiotracers into the clinical practice has resulted in the significant improvement of accuracy in the detection of prostate carcinoma (PCa). PSMA-expression in ganglia has been regarded as an important pitfall in prostate carcinoma-PET diagnostics but has not found any practical use for diagnosis or therapy. METHODS: We explored this phenomenon and demonstrated the applicability of peripheral ganglia in healthy rats as surrogates for small PSMA positive lesions for the preclinical evaluation of diagnostic PCa PET probes. Healthy rats were measured with PET/CT using the tracers [18F]DCFPyL, [Al18F]PSMA-11 and [68Ga]PSMA-11. Sections of ganglia were stained with an anti-PSMA antibody. [18F]DCFPyL uptake in ganglia was compared to that in LNCaP tumor xenografts in mice. RESULTS: Whereas [18F]DCFPyL and [68Ga]PSMA-11 were stable in vivo and accumulated in peripheral ganglia, [Al18F]PSMA-11 suffered from fast in vivo deflourination resulting in high bone uptake. Ganglionic PSMA expression was confirmed by immunohistochemistry. [18F]DCFPyL uptake and signal-to-noise ratio in the superior cervical ganglion was not significantly different from LNCaP xenografts. CONCLUSIONS: Our results demonstrated the non-inferiority of the novel model compared to conventionally used tumor xenografts in immune compromised rodents with regard to reproducibility and stability of the PSMA signal. Furthermore, the model involves less expense and efforts while it is permanently available and avoids tumor-growth associated animal morbidity and distress. To the best of our knowledge, this is the first tumor-free model suitable for the in vivo evaluation of tumor imaging agents.


Assuntos
Antígenos de Superfície/metabolismo , Gânglios/diagnóstico por imagem , Glutamato Carboxipeptidase II/metabolismo , Lisina/análogos & derivados , Glicoproteínas de Membrana/metabolismo , Compostos Organometálicos/metabolismo , Tomografia Computadorizada com Tomografia por Emissão de Pósitrons , Neoplasias da Próstata/diagnóstico por imagem , Compostos Radiofarmacêuticos/metabolismo , Ureia/análogos & derivados , Animais , Autorradiografia , Gânglios/metabolismo , Gânglios Espinais/metabolismo , Lisina/metabolismo , Masculino , Camundongos , Camundongos SCID , Especificidade de Órgãos , Neoplasias da Próstata/metabolismo , Ratos , Ratos Long-Evans , Ratos Wistar , Gânglio Trigeminal/metabolismo , Ureia/metabolismo
11.
J Pediatr Surg ; 54(11): 2318-2324, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31079866

RESUMO

BACKGROUND: Retained transition zone is a leading cause of obstructive symptoms after pull-through operation in Hirschsprung's disease. OBJECTIVE: We aimed to evaluate the extent of the histological transition zone in patients with Hirschsprung's disease. DESIGN: We performed an observational study. DAB+ immunohistochemistry for Protein Gene Product 9.5 was used to evaluate the neuronal networks in serial sections of pull-through specimens obtained from children with Hirschsprung's disease (n = 12). Reference ranges for ganglion size/density and nerve trunk diameter were statistically determined using healthy controls obtained from colostomy specimens from children with anorectal malformations (n = 8). The transition zone was defined as ganglionic bowel exhibiting ganglion hypoplasia, hypertrophic nerve trunks, or partial circumference aganglionosis. RESULTS: The mean submucosal nerve trunk diameter in controls was 19.56 µm +/- 3.87 µm. The median age at pull-through for Hirschsprung's disease was 5 months (3-14 months). The median length of the transition zone across the population was 8 cm (4-22 cm). Median transition zone extent was significantly longer in patients with long-segment aganglionosis (n = 6) compared to rectosigmoid aganglionosis (n = 6, 13 cm vs 6 cm, p = 0.041). Due to the age of the patients enrolled, long-term follow-up of bowel function is not yet available. CONCLUSION: Our data suggest that, in children with rectosigmoid Hirschsprung's disease, the transition zone can extend for up to 13 cm. In children with long-segment disease, a longer transition zone is possible. Extended resection at a minimum 5 cm beyond the most distal ganglionic intra-operative biopsy and intra-operative histological examination of the proximal resection margin are required to minimize transition zone pull-through. LEVEL OF EVIDENCE: 2.


Assuntos
Doença de Hirschsprung , Procedimentos Cirúrgicos do Sistema Digestório , Gânglios/patologia , Gânglios/fisiopatologia , Doença de Hirschsprung/patologia , Doença de Hirschsprung/fisiopatologia , Doença de Hirschsprung/cirurgia , Humanos , Lactente , Complicações Pós-Operatórias
12.
Pol J Vet Sci ; 22(1): 151-155, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30997776

RESUMO

The present study investigated the expression of androgen receptor (AR) in neurons of the anterior pelvic ganglion (APG) and celiac-superior mesenteric ganglion (CSMG; ganglion not involved in the innervation of reproductive organs) in the male pig with quantitative real-time PCR (qPCR) and immunohistochemistry. qPCR investigations revealed that the level of AR gene expression in the APG tissue was approximately 2.5 times higher in the adult (180-day-old) than in the juvenile (7-day-old) boars. Furthermore, in both the adult and juvenile animals it was sig- nificantly higher in the APG than in CSMG tissue (42 and 85 times higher, respectively). Immu- nofluorescence results fully confirmed those obtained with qPCR. In the adult boars, nearly all adrenergic (DßH-positive) and the majority of non-adrenergic neurons in APG stained for AR. In the juvenile animals, about half of the adrenergic and non-adrenergic neurons were AR-posi- tive. In both the adult and juvenile animals, only solitary CSMG neurons stained for AR. The present results suggest that in the male pig, pelvic neurons should be considered as an element of highly testosterone-dependent autonomic circuits involved in the regulation of urogenital func- tion, and that their sensitization to androgens is a dynamic process, increasing during the prepu- bertal period.


Assuntos
Gânglios/metabolismo , Plexo Hipogástrico/fisiologia , Mesentério/inervação , Receptores Androgênicos/metabolismo , Suínos/metabolismo , Animais , Regulação da Expressão Gênica/fisiologia , Imuno-Histoquímica , Masculino
13.
Genes (Basel) ; 10(4)2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30987054

RESUMO

Genetic information of reproduction and growth is essential for sustainable molluscan fisheries and aquaculture management. However, there is limited knowledge regarding the reproductive activity of the commercially important Pacific abalone Haliotis discus hannai. We performed de novo transcriptome sequencing of the ganglia in sexually immature and mature female Pacific abalone to better understand the sexual maturation process and the underlying molecular mechanisms. Of the ~305 million high-quality clean reads, 76,684 transcripts were de novo-assembled with an average length of 741 bp, 28.54% of which were annotated and classified according to Gene Ontology terms. There were 256 differentially expressed genes between the immature and mature abalone. Tandem mass spectrometry analysis, as compared to the predicted-peptide database of abalone ganglia transcriptome unigenes, identified 42 neuropeptide precursors, including 29 validated by peptidomic analyses. Label-free quantification revealed differential occurrences of 18 neuropeptide families between immature and mature abalone, including achatin, FMRFamide, crustacean cardioactive peptide, and pedal peptide A and B that were significantly more frequent at the mature stage. These results represent the first significant contribution to both maturation-related transcriptomic and peptidomic resources of the Pacific abalone ganglia and provide insight into the roles of various neuropeptides in reproductive regulation in marine gastropods.


Assuntos
Gânglios/metabolismo , Gastrópodes/genética , Reprodução/genética , Transcriptoma/genética , Animais , Feminino , Gânglios/crescimento & desenvolvimento , Gastrópodes/crescimento & desenvolvimento , Regulação da Expressão Gênica/genética , Ontologia Genética , Neuropeptídeos/genética , Maturidade Sexual/genética , Espectrometria de Massas em Tandem
14.
BMC Pediatr ; 19(1): 93, 2019 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-30953480

RESUMO

BACKGROUND: Total colonic and small bowel aganglionosis (TCSA) occurs in less than 1% of all Hirschsprung's disease patients. Currently, the mainstay of treatment is surgery. However, in patients with TCSA, functional outcomes are often poor. A characteristic transition zone in TCSA can be difficult to identify which may complicate surgery and may often require multiple operations. CASE PRESENTATION: We present the case of a male infant who was diagnosed with biopsy-proven total colonic aganglionosis with extensive small bowel involvement as a neonate. The patient was diverted at one month of age based on leveling biopsies at 10 cm from the Ligament of Treitz. At 7 months of age, during stoma revision for a prolapsed stoma, intra-operative peristalsis was observed in nearly the entire length of the previously aganglionic bowel, and subsequent biopsies demonstrated the appearance of mature ganglion cells in a previously aganglionic segment. CONCLUSIONS: TCSA remains a major challenge for pediatric surgeons. Our case introduces new controversy to our understanding of aganglionosis. Our observations warrant further research into the possibility of post-natal ganglion maturation and encourage surgeons to consider a more conservative surgical approach.


Assuntos
Gânglios/patologia , Doença de Hirschsprung/cirurgia , Intestino Delgado/inervação , Biópsia , Colo/anormalidades , Colo/patologia , Colo/cirurgia , Doença de Hirschsprung/patologia , Humanos , Recém-Nascido , Enteropatias/diagnóstico por imagem , Enteropatias/patologia , Intestino Delgado/diagnóstico por imagem , Intestino Delgado/patologia , Intestino Delgado/cirurgia , Masculino , Radiografia
15.
Environ Sci Pollut Res Int ; 26(14): 14461-14472, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30868461

RESUMO

Rotenone (ROT) was shown to affect cerebral ganglions (CGs) of Lumbricus terrestris as a pioneering observation in our earlier investigation. Though ROT is a well-known neurotoxin causing neurodegeneration (ND), the precipitation of movement dysfunction remains largely unknown. We have designed the current study to analyze motor abnormalities in worms by exposing them to different concentrations (0.0-0.4 ppm) of ROT for 7 days. GABA, cholinergic receptor, serotonin transporter (SERT), acetylcholine esterase (AchE), and dopamine-ß-hydroxylase that are well known for their involvement in neuromuscular junctions were investigated by qRT-PCR. Further, neuronal mitochondrial genes (cytochrome C oxidase-2, NADH deydrogenase-1, cytochrome-b) and actin-1 that are essential for regeneration and calreticulin (phagocytosis) were investigated. The levels of neurotransmitters, lipids, ATPase, neuronal behavior analyses, and fluorescence analysis (lipid droplets) were performed in CGs which showed significant variations at 0.3 ppm. Ultrastructural changes in lipid droplet and neuromelatonin were prominent in 0.3 ppm. Dose-dependent effect of ROT on behavior alteration and expression of m-RNAs studied suggested that at 0.3 ppm, it could deteriorate motor and cognitive functions. We predict that perhaps, by virtue of its effect on cerebral ganglionic genes and their neurotransmitting potential, ROT may cause morbidities that resemble features characteristic of hemiparkinsonic degeneration.


Assuntos
Neurônios/efeitos dos fármacos , Neurotoxinas/toxicidade , Oligoquetos/fisiologia , Rotenona/toxicidade , Animais , Cognição/efeitos dos fármacos , Gânglios/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Neurotransmissores/metabolismo , Oligoquetos/efeitos dos fármacos , RNA Mensageiro/metabolismo , Poluentes do Solo/toxicidade , Testes de Toxicidade
16.
Methods Mol Biol ; 1952: 117-125, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30825171

RESUMO

The cellular components of the enteric nervous system (ENS), namely enteric neurons and glia, display plasticity and respond to environmental cues deriving from growth factors, extracellular matrix (ECM) molecules, and cell-surface molecules, both in physiological and pathological conditions. ECM, in particular, provides an important framework for the enteric microenvironment and influences the homeostasis of myenteric neuronal circuitries. Isolation of pure myenteric plexus preparations from adult tissue permits to investigate changes in the ENS involving specific ECM, such as hyaluronan. This approach is based upon the possibility to isolate myenteric ganglia from the intestinal wall of either adult animals or humans, after microdissection and subsequent enzymatic digestion of the tissue. Enteric ganglia are free of connective tissue, extracellular collagen, and blood vessels, and thus treatment of intact intestinal segments with highly purified collagenases permits ganglia isolation from the surrounding smooth muscle cells. In this chapter, we describe methods for visualizing HA in isolated primary cultures of adult rat small intestine myenteric ganglia.


Assuntos
Imunofluorescência/métodos , Gânglios/química , Ácido Hialurônico/análise , Intestino Delgado/inervação , Microscopia Confocal/métodos , Plexo Mientérico/química , Animais , Células Cultivadas , Gânglios/citologia , Gânglios/ultraestrutura , Plexo Mientérico/citologia , Plexo Mientérico/ultraestrutura , Ratos , Técnicas de Cultura de Tecidos/métodos
17.
Cell Tissue Res ; 376(3): 471-484, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30778730

RESUMO

Neuropeptide F in invertebrates is a homolog of neuropeptide Y in mammals and it is a member of FMRFamide-related peptides. In arthropods, such as insects, there are two types of neuropeptide F comprising long neuropeptide F (NPF) and short neuropeptide F (sNPF). Both NPFs are known to play a crucial role in the regulations of foraging, feeding-related behaviors, circadian rhythm, stress responses, aggression and reproduction in invertebrates. We have earlier found that in the giant freshwater prawn, Macrobrachium rosenbergii, there are three isoforms of NPF and four isoforms of sNPF and that NPFs are expressed in the eyestalks and brain. In the present study, we investigate further the tissue distribution of NPF-I in the ventral nerve cord (VNC) and its role in the development of testes in small male (SM) Macrobrachium rosenbergii. By immunolocalization, using the rabbit polyclonal antibody against NPF-I as a probe, we could detect NPF-I immunoreactivity in the neuropils and neuronal clusters of the subesophageal ganglia (SEG), thoracic ganglia (TG) and abdominal ganglia (AG) of the SM prawns. In functional assays, the administrations of synthetic NPF-I (KPDPTQLAAMADALKYLQELDKYYSQVSRPRFamide) and sNPF (APALRLRFamide) peptides significantly increased the growth rates of SM prawns and significantly increased the gonadosomatic index (GSI) and proliferations of early germ cells in the seminiferous tubules of their testes. It is, therefore, suggestive that NPFs may play critical roles in energy homeostasis towards promoting growth as well as testicular development in prawns that could be applied in the aquaculture of this species.


Assuntos
Gânglios/metabolismo , Células Germinativas/crescimento & desenvolvimento , Neuropeptídeos/metabolismo , Neurópilo/metabolismo , Palaemonidae/crescimento & desenvolvimento , Testículo/crescimento & desenvolvimento , Animais , Proliferação de Células , Masculino , Palaemonidae/metabolismo
18.
J Sex Med ; 16(1): 27-41, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30621923

RESUMO

BACKGROUND: Erectile dysfunction (ED) is common following radiation therapy (RT) for prostate cancer. Although the cause of RT-induced ED is unknown, damage to both the neuronal and vascular components supporting erections are often implicated. AIM: To determine the effects of prostatic RT on erections, penile vascular physiology, and major pelvic ganglia (MPG) neuron growth and survival in a rat model. METHODS: Male rats underwent 0 Gy or 22 Gy single fraction of prostate-confined, conformal RT. At 2 weeks or 10 weeks post-RT (n = 10/group), cavernous nerve stimulation was performed and erections were assessed. Tissue bath experiments were performed to assess both penile artery and internal pudendal artery (IPA) function. MPGs were dissociated and neurons grown in culture for 72 hours. Immunofluorescence staining was done to quantify neuron survival (terminal deoxynucleotidyl transferase nick-end labeling), outgrowth (beta-tubulin III), type (nitric oxide synthase [nNOS] and tyrosine hydroxylase [TH]), and nerve injury markers (small GTPase Rac1 and ninjurin-1 [Ninj-1]). Whole MPG real-time quantitative polymerase chain reaction (qPCR) was performed to measure expression of genes related to nerve type, neuron injury, repair, and myelination, such as Ninj-1, Rac1, ATF3, GAP43, GFAP, SOX10, and KROX20. OUTCOMES: Intracavernosal pressure (ICP) to mean arterial pressure (MAP) ratio, smooth muscle contractility and relaxation, gene expression, neuritogenesis, and apoptosis. RESULTS: Following RT, ICP/MAP was unchanged at 2 weeks or 10 weeks. Nerve-mediated penile contraction was increased at 2 weeks, whereas adrenergic contraction was reduced at 10 weeks. Penile relaxation and IPA vasoreactivity were unchanged. Neuronal apoptosis was more than doubled both early and late post-RT. RT caused a progressive decrease in neurite branching but an early increase and then late decrease in neurite lengthening. RT reduced the numbers of nNOS-positive neurons both early and late and also decreased MPG nitrergic gene expression. TH neurons and gene expression were unchanged at 2 weeks; however, both were decreased after 10 weeks. Although most markers of gene injury and repair were unaffected early post-RT, MPG expression of Ninj1 and GFAP increased. After 10 weeks, Ninj1 and GFAP remained elevated while markers of neuron injury (ATF3), outgrowth (GAP43 and Rac1), and myelin regulation (SOX10) were decreased. CLINICAL TRANSLATION: RT-induced ED may result from damage to the ganglia controlling erections. STRENGTHS & LIMITATIONS: This study used a clinically relevant, prostate-confined model to examine neurovascular structures not accessible in human studies. Unfortunately, rats did not exhibit ED at this time point. CONCLUSION: This is the first study to demonstrate impaired health and regeneration potential of dissociated MPG neurons following RT. Neuronal injury was apparent early post-RT and persisted or increased over time but was insufficient to cause ED at the time points examined. Powers SA, Odom MR, Pak ES, et al. Prostate-Confined Radiation Decreased Pelvic Ganglia Neuronal Survival and Outgrowth. J Sex Med 2019;16:27-41.


Assuntos
Disfunção Erétil/etiologia , Ereção Peniana/efeitos da radiação , Neoplasias da Próstata/radioterapia , Animais , Modelos Animais de Doenças , Gânglios/metabolismo , Plexo Hipogástrico/metabolismo , Masculino , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Pênis/fisiopatologia , Ratos , Ratos Sprague-Dawley , Traumatismos do Sistema Nervoso/complicações , Tirosina 3-Mono-Oxigenase/metabolismo
19.
Acta Histochem ; 121(2): 234-239, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30616841

RESUMO

The gonadotropin-releasing hormone (GnRH) gene sequence has been identified in an annelid polychaete marine worm using continual genome sequencing. The distribution of GnRH immunoreactive (ir) cell bodies and fibers in the nerve ganglion of the clam worm Perinereis aibuhitensis (Polychaeta) was examined by immunohistochemistry using a newly produced rabbit polyclonal antibody raised against the marine worm GnRH (mwGnRH). The specificity of the antibody was confirmed by dot blot assay. The antibody cross-reacted with mwGnRH, but not with other forms of GnRH such as octopus GnRH, tunicate GnRH-I, II, owl limpet GnRH, and lamprey GnRH-II. In P. aibuhitensis, mwGnRH-ir cell bodies were detected in the nuclei 15-22, the caudal part of the cerebral ganglion. Furthermore, mwGnRH-ir fibers were mainly observed in the optic neuropil, but mwGnRH-ir fibers were also detected in the central neuropil region, the subpharyngeal ganglion, and the ventral nerve cord. These results indicate that mwGnRH is synthesized in the cerebral ganglion, is transported through the subpharyngeal ganglion and the ventral nerve cord, and functions either as a neurotransmitter or neuromodulator.


Assuntos
Corpo Celular/fisiologia , Gânglios/metabolismo , Hormônio Liberador de Gonadotropina/metabolismo , Imuno-Histoquímica , Animais , Encéfalo/metabolismo , Imuno-Histoquímica/métodos , Fibras Nervosas/metabolismo , Poliquetos , Coelhos
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