Morphological and neurochemical characterization of sensory neurons and motorneurons innervating the urinary bladder tringone and urethral muscle in porcine

Il trigono della vescica urinaria (UBT) è un'area limitata attraverso la quale penetrano nella vescica la maggior parte dei vasi e fibre e in cui le fibre nervose e neuroni intramurali sono più concentrati. Mediante l’utilizzo combinato di un tracciante retrogrado(FB) e dell’immunoistochimica sono stati valutati il fenotipo e l’area del soma dei neuroni dei gangli spinali (DRG), dei neuroni post-gangliari, il fenotipo dei gangli della catena simpatica (STG) e i gangli mesenterici caudali (CMG) innervanti l’UBT. - Caratterizzazione dei neuroni dei DRG con: peptide correlato al gene della calcitonina (CGRP)(30±3%, 29±3%, rispettivamente), sostanza P(SP)(26±8%, 27±12%), ossido nitrico sintasi neuronale (nNOS)(21±4%; 26±7%), neurofilamento 200kDa (NF200)(75±14%, 81±7% ) , transient receptor potential vanilloid1 (TRPV1)(48±13%, 43±6%) e isolectina-B4-positivi (IB4) (56±6%;43±10%). I neuroni sensoriali, distribuiti da L2 a Ca1 (DRG), hanno presentato una localizzazione segmentale, mostrando maggior densità nei DRG L4-L5 e S2-S4. I neuroni sensoriali lombari sono risultati significativamente più grandi di quelle sacrali (1.112±624μm2 vs716±421μm2). Complessivamente, questi dati indicano che le vie lombari e sacrali probabilmente svolgono ruoli diversi nella trasmissione sensitiva del trigono della vescica urinaria. -I neuroni FB+ della STG e dei CMG sono risultati immunoreattivi per la tirosina idrossilasi (TH)(66±10,1%, 53±8,2%, rispettivamente), la dopamina beta-idrossilasi (DβH)(62±6,2%, 52±6,2%), neuropeptideY (NPY)(59±8%; 66±7%), CGRP(24±3%, 22±3%), SP(22±2%; 38±8%), polipeptide intestinale vasoattivo (VIP)(19±2%; 35±4%), nNOS(15±2%; 33±8%), trasportatore vescicolare dell'acetilcolina (VAChT)(15±2%; 35±5%), leu-encefalina (LENK)(14±7%; 26±9%), e somatostatina (SOM)(12±3%;32±7%).Il numero medio di neuroni FB+ (1845,1±259,3) era nella STG in L1-S3, con i pirenofori più piccoli (465,6±82.7μm2). Un gran numero (4287,5±1450,6) di neuroni FB+ di piccole dimensioni (476,1±103,9μm2) sono stati localizzati lungo il margine dei CMG. Il maggior numero (4793,3±1990,8) di neuroni FB + è stato osservato nel plesso pelvico, dove i neuroni marcati erano raggruppati in micro-gangli e con pirenoforo ancora più piccolo (374,9±85,4 μm2).The urinary bladder trigone (UBT) is a limited area through which the majority of vessels and nerve fibers penetrate into the urinary bladder and where nerve fibers and intramural neurons are more concentrated. The phenotype and soma cross-sectional area of dorsal root ganglion (DRG) neurons, the extramural post-ganglionic autonomic neurons, the phenotype of sympathetic trunk ganglia (STG) neurons and caudal mesenteric ganglia (CMG) neurons innervating the porcine UBT were evaluated by coupling retrograde tracer technique and immunohistochemistry. -Porcine lumbosacral DRG neurons were characterized neurochemically: calcitonin gene-related peptide (CGRP)(30±3%; 29±3%, respectively), substance P (SP)(26±8%; 27±12%), neuronal nitric oxide synthase (nNOS)(21±4% and; 26±7%), neurofilament200kDa (NF200)(75±14%; 81±7%), and transient receptor potential vanilloid1 (TRPV1)(48±13%; 43±6%), and labeled for isolectin-B4 (IB4)(56±6%; 43±10%). UBT sensory neurons, which were distributed from L2 to Ca1 DRG, had a segmental localization, showing their highest density in L4–L5 and S2–S4 DRG. The lumbar UBT sensory neurons were significantly larger than sacral ones (1,112±624µm2 vs. 716±421µm2). Taken together, these data indicate that the lumbar and sacral pathways probably play different roles in sensory transmission from the UBT. -STG and CMG FB+ neurons were IR for tyrosine hydroxylase (TH)(66±10.1%; 52.7±8.2%, respectively), dopamine beta-hydroxylase (DβH)(62±6.2%; 52±6.2%), neuropeptide Y (NPY)(59±8%; 66±7%), CGRP (24±3%; 22±3%), SP (22±2%; 38±8%), vasoactive intestinal polypeptide (VIP)(19±2%; 35±4%), nNOS (15±2%; 33±8%), vesicular acetylcholine transporter (VAChT)(15±2%; 35±5%), leuenkephalin (LENK)(14±7%; 26±9%), and somatostatin (SOM)(12±3%; 32±7%). A mean number of 1845.1±259.3 FB+ neurons were localized in the L1-S3 STG, which appeared as small pericarya (465.6±82.7µm2). A large number (4287.5±1450.6) of small (476.1±103.9µm2) FB+ neurons were localized mainly along a border of both CMG. The largest number (4793.3±1990.8) of FB+ neurons was observed in the pelvic plexus (PP), where labeled neurons were often clustered within different microganglia and had smaller soma cross-sectional area (374.9±85.4 µm2)

Expression of \u3b22 adrenoceptors within enteric neurons of the horse ileum

The activity of the gastrointestinal tract is regulated through the activation of adrenergic receptors (ARs). Since data concerning the distribution of ARs in the horse intestine is virtually absent, we investigated the distribution of \u3b22-AR in the horse ileum using double-immunofluorescence. The \u3b22-AR-immunoreactivity (IR) was observed in most (95%) neurons located in submucosal plexus (SMP) and in few (8%) neurons of the myenteric plexus (MP). Tyrosine hydroxylase (TH)-IR fibers were observed close to neurons expressing \u3b22-AR-IR. Since \u3b22-AR is virtually expressed in most neurons located in the horse SMP and in a lower percentage of neurons in the MP, it is reasonable to retain that this adrenergic receptor could regulate the activity of both secretomotor neurons and motor neurons innervating muscle layers and blood vessels. The high density of TH-IR fibers near \u3b22-AR-IR enteric neurons indicates that the excitability of these cells could be directly modulated by the sympathetic system

Mucosal mRNA Cytokines’ Profile of Gastric Wall in Neonatal Foals: Comparison with Endoscopy and Histology

Gastritis and gastric ulcerations occur frequently in neonatal foals. The relationship between cytokines expressed by gastric mucosa and gastric histopathology in healthy or sick foals has never been investigated. The aim of this study was to compare the histological diagnosis and endoscopic view with cytokine expression (TNF-a, IL-1b, IL-4, IL-8, IL-13, and IFN-g) of gastric mucosa. Twenty-two foals were definitively enrolled in the study: 19 were critically ill, and 3 were healthy foals. Gastric biopsy specimens were collected for histological examination and for cytokine mRNA qualitative real-time PCR analysis. This study shows that there is a substantial agreement between histology and endoscopy and that foals with evidence of gastritis and gastric ulcerations have higher probability of expressing TNF-a. Moreover, the overall profile of cytokines expression, with a low percentage of IFN-g, a high percentage of IL-4, and the absence of IL-13, suggests a down-regulation of the Th1 cell-mediated immune response and an impaired Th2 response in the gastric wall in the neonatal period

Neurochemical features of boar lumbosacral dorsal root ganglion neurons and characterization of sensory neurons innervating the urinary bladder trigone

Porcine lumbosacral dorsal root ganglion (DRG) neurons were neurochemically characterized by using six neuronal markers: calcitonin gene-related peptide (CGRP), substance P (SP), neuronal nitric oxide synthase (nNOS), neurofilament 200kDa (NF200), transient receptor potential vanilloid 1 (TRPV1), and isolectin B4 (IB4) from Griffonia simplicifolia. In addition, the phenotype and cross-sectional area of DRG neurons innervating the urinary bladder trigone (UBT) were evaluated by coupling retrograde tracer technique and immunohistochemistry. Lumbar and sacral DRG neuronal subpopulations were immunoreactive (IR) for CGRP (30 \ub1 3% and 29 \ub1 3%, respectively), SP (26 \ub1 8% and 27 \ub1 12%, respectively), nNOS (21 \ub1 4% and 26 \ub1 7%, respectively), NF200 (75 \ub1 14% and 81 \ub1 7%, respectively), and TRPV1 (48 \ub1 13% and 43 \ub1 6%, respectively), and labeled for IB4 (56 \ub1 6% and 43 \ub1 10%, respectively). UBT sensory neurons, which were distributed from L2 to Ca1 DRG, had a segmental localization, showing their highest density in L4-L5 and S2-S4 DRG. Lumbar and sacral UBT sensory neurons expressed similar percentages of NF200 immunoreactivity (64 \ub1 33% and 58 \ub1 12%, respectively) but showed a significantly different immunoreactivity for CGRP, SP, nNOS, and TRPV1 (56 \ub1 9%, 39 \ub1 15%, 17 \ub1 13%, 62 \ub1 10% vs. 16 \ub1 6%, 16 \ub1 11%, 6 \ub1 1%, 45 \ub1 24%, respectively). Lumbar and sacral UBT sensory neurons also showed different IB4 labeling (67 \ub1 19% and 48 \ub1 16, respectively). Taken together, these data indicate that the lumbar and sacral pathways probably play different roles in sensory transmission from the UBT. The findings related to cell size also reinforced this hypothesis, because lumbar UBT sensory neurons were significantly larger than sacral ones (1,112 \ub1 624 \u3bcm(2) vs. 716 \ub1 421 \u3bcm(2) )