Browsing by Subject "Innervation"
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- PublicationOpen AccessAcetylcholinesterase-positive and paraformaldehyde-induced-fluorescence-positive innervation in the upper eyelid of the sheep (Ovis aries)(Murcia : F. Hernández, 2001) Aisa, J.; Lahoz, M.; Serrano, P.; Perez-Castejon, M.C.; Junquera, C.; Martínez Ciriano, M.C.; Pes, N.; Vera Gil, A.This is the first study which describes the innervation of some eyelid structures, such as the glands of Moll and the glands of Zeiss. It is also the first to investigate the innervation pattern of the eyelid as a whole. We have studied the acetylcholinesterase-positive and paraformaldehyde-induced-fluorescence-positive (FIF+) innervation pattern of the different structures that constitute the upper eyelid of the sheep. There is widespread acetylcholinesterase-positive innervation in the epithelium, but not such an abundant FIF+ innervation. Both types of innervation are represented in the connective tissue by trunks or fibers that are distributed towards the different structures immersed within them. In the glands of Zeiss, cholinesterasepositive innervation is much more widespread than FIF innervation. On the contrary, the glands of Moll present denser FIF+ innervation than acetylcholinesterasepositive innervation. The Meibomian glands and the lachrymal glands show a rich acetylcholinesterasepositive and FIF+ innervation. Eyelid muscle innervation is mainly acetylcholinesterase-positive. In the conjunctive membrane there is no acetylcholinesterase-positive innervation, and only scarce FIF+ fibers can be demonstrated.
- PublicationOpen AccessAngiotensinergic innervation of the kidney: Localization and relationship with catecholaminergic postganglionic and sensory nerve fibers(F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2012) Bohlender, Jürgen; Pfarrer, Beat; Patil, Jaspal; Nussberger, Jürg; Thalmann, Georg N. Thalmann; Imboden, HansWe describe an angiotensin (Ang) II-containing innervation of the kidney. Cryosections of rat, pig and human kidneys were investigated for the presence of Ang II-containing nerve fibers using a mouse monoclonal antibody against Ang II (4B3). Co-staining was performed with antibodies against synaptophysin, tyrosine 3-hydroxylase, and dopamine beta-hydroxylase to detect catecholaminergic efferent fibers and against calcitonin gene-related peptide to detect sensory fibers. Tagged secondary antibodies and confocal light or laser scanning microscopy were used for immunofluorescence detection. Ang II-containing nerve fibers were densely present in the renal pelvis, the subepithelial layer of the urothelium, the arterial nervous plexus, and the peritubular interstitium of the cortex and outer medulla. They were infrequent in central veins and the renal capsule and absent within glomeruli and the renal papilla. Ang II-positive fibers represented phenotypic subgroups of catecholaminergic postganglionic or sensory fibers with different morphology and intrarenal distribution compared to their Ang II-negative counterparts. The Ang II-positive postganglionic fibers were thicker, produced typically fusiform varicosities and preferentially innervated the outer medulla and periglomerular arterioles. Ang II-negative sensory fibers were highly varicose, prevailing in the pelvis and scarce in the renal periphery compared to the rarely varicose Ang II-positive fibers. Neurons within renal microganglia displayed angiotensinergic, cate-cholaminergic, or combined phenotypes. Our results suggest that autonomic fibers may be an independent source of intrarenal Ang II acting as a neuropeptide co-transmitter or neuromodulator. The angiotensinergic renal innervation may play a distinct role in the neuronal control of renal sodium reabsorption, vasomotion and renin secretion.
- PublicationOpen AccessAnterior cruciate ligament innervation in primary knee osteoarthritis(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2022) Guerra González, Adrián; Casa, Carmen da; Crespo, Íñigo; Pescador, David; Benito Garzón, Lorena; Blanco, Juan FObjective. To relate the Anterior Cruciate Ligament (ACL) innervation and histologic degeneration status to the knee osteoarthritis radiologic and functional status. Design. Prospective observational study including 30 consecutive patients affected by primary knee osteoarthritis undergoing Total Knee Arthroplasty (TKA). All patients suffering secondary knee osteoarthritis, an antecedent of an infectious process, malignant process, autoimmune disorder, or previous knee surgery were excluded. We recorded biodemographic, clinical, and radiologic variables of all participants previous to the TKA procedure. ACL tissue was harvested during TKA standard procedure and the obtained sample was fixed in 4% formalin and paraffinembedded. ACL cross-sections were stained by haematoxylin-eosin and Gallego staining for elastic and collagen fibers, and Sevier-Munger silver staining for nervous tissue. Results. ACL samples histologic degeneration classification reported 15.4% normal, 23.1% slight, 26.9% mild, 11.5% moderate and 23.1% marked. We noted 46.2% large nervous fascicles, 15.4% medium fascicles, 3.8% small fascicles, and no nerve fibers were found in 34.6% ACL samples. No significant correlation was found between the histologic degeneration and the nervous fiber quantification (p>0.05, in all cases). We noted a significant histologic degeneration inverse correlation with the VAS scale (p=0.016), and nervous fiber quantification correlation with Lequesne maximum distance walked punctuation (p=0.043). We also noted greater nervous fiber quantification with minor radiological knee osteoarthritis (Kellgren-Lawrence grade II). Conclusions. ACL degeneration and innervation deficit may play a role in primary knee osteoarthritis onset, but the lack of a defining relationship among the different parameters assessed justifies further research in greater populations.
- PublicationOpen AccessInnervation and nerve-immune cell contacts in mouse Peyer's patches(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Al Shalan, Huda A.M.; Hu, Dailun; Greene, Wayne K.; Ma, BinNeural regulation of the function of the gastrointestinal tract (GIT) relies on a delicate balance of the two divisions of its nervous system, namely, the intrinsic and extrinsic divisions. The intrinsic innervation is provided by the enteric nervous system (ENS), whereas the extrinsic innervation includes sympathetic/parasympathetic nerve fibers and extrinsic sensory nerve fibers. In the present study, we used immunofluorescent staining of neurofilament-heavy (NF-H) to reveal the distribution of nerve fibers and their associations with immune cells inside mouse Peyer’s patches (PP), an essential part of gut-associated lymphoid tissue (GALT). Our results demonstrate (1) the presence of an extensive meshwork of NF-H- immunoreactive presumptive nerve fibers in all PP compartments including the lymphoid nodules, interfollicular region, follicle-associated epithelium, and subepithelial dome; (2) close associations/contacts of nerve fibers with blood vessels including high endothelial venules, indicating neural control of blood flow and immune cell dynamics inside the PP; (3) close contacts between nerve fibers/endings and B/T cells and various subsets of dendritic cells (e.g., B220 - , B220 + , CD4 - , CD4 + , CD8 - , and CD8 + ). Our novel findings concerning PP innervation and nerve-immune cell contacts in situ should facilitate our understanding of bi- directional communications between the PNS and GALT. Since the innervation of the gut, including PP, might be important in the pathogenesis and progression of some neurological, infectious, and autoimmune diseases, e.g., prion diseases and inflammatory bowel disease, better knowledge of PNS-immune system interactions in the GALT (including PP) should benefit the development of potential treatments for these diseases via neuroimmune manipulations.
- PublicationOpen AccessInvestigation of general and cytoskeletal markers to estimate numbers and proportions of neurons in the human intestine(Murcia : F. Hernández, 2006) Ganns, D.; Schrödl, F.; Neuhuber, W.; Brehmer, A.An important requirement in pathological diagnostics in the human enteric nervous system (ENS) is the estimation of the total numbers of neurons and of proportions of distinct subpopulations. In this study, we compared the suitability of two suggested panneuronal markers, cuprolinic blue (CB) and anti-Hu-protein (HU), for staining and counting human myenteric neurons in wholemounts, derived from small and large intestinal samples. Furthermore, the proportional expression of three cytoskeletal intermediate filaments, a-internexin (IN), neurofilament 200 (NF) and peripherin (PE), was correlated with both CB and HU. In 8 CB- and HUstained wholemounts, 93.3% of all neurons were double labeled, 3.3% of neurons were stained only with CB whereas 3.3% were immuno-stained only for HU. Thus, both markers were comparably reliable in representing the putative total human myenteric neuron population in our material. The wholemounts double stained for IN/CB or IN/HU revealed between 56.2 and 71.5% of neurons to be IN-reactive. Between 42.8 and 50.9% of neurons were immunoreactive for NF whereas 53.9 to 62.4% of neurons were reactive for PE. Although our sample number was too small to allow final conclusions, we suggest that the variations in proportions of intermediate filament expression we observed may be due to individual circumstances rather than to correlation with age or region. The proportions of neurons positive for IN, NF or PE but unstained by CB histochemical or HU immunohistochemical techniques was between 0 and 2.2%. We conclude that both CB and HU techniques are suitable methods for representation of almost all myenteric neurons in the human gut and that the differential expression of the cytoskeletal proteins investigated has to be included in the classification of enteric neurons in pathological diagnostics of human gastrointestinal diseases.
- PublicationOpen Accesslmmunocytochemical study on the innervation of the chicken pancreas by vasoactive intestinal polypeptide (VIP)-containing nerves(Murcia : F. Hernández, 1997) Hiramatsu, K.; Oshima, K.Ultrastructural localization of vasoactive intestinal polypeptide (VIP) was studied in the chicken pancreas by the immunocytochemical method. VIPimmunoreactive nerve endings were found in contact with acinar cells in the pericapillary space or between acinar cells. They also terminated on epithelial cells of arteriolae. In pancreatic islets, VIP-immunoreactive nerve endings were in contact with B-cells, but not with A- or D-cells. VIP-imrnunoreactive terminals contained many small clear vesicles which indicate the cholinergic feature. Nerve endings containing many small clear vesicles but not showing VIP-immunoreactivity were also detected in both the exocrine and endocrine pancreas. According to these data and the previous studies concerning the cholinergic and VIPergic innervation of the chicken pancreas, it is suggested that VIP-containing cholinergic nerves innervate B-cells and cholinergic nerves not containing VIP innemate D-cell.
- PublicationOpen AccessMorphological differences among nerve fiber endings in the rat oral mucosa as revealed by methylene blue staining(Murcia : F. Hernández, 1996) Müller, T.The nerve fiber distribution in the oral mucosa of the soft palate and palatoglossal arch of the rat was studied by means of methylene blue supravital staining. it was focussed primarily on the dye uptake of intraepithelial nerve fibers. Differences in the morphology of nerve fiber terminations were found between these regions of the oral mucosa. In the soft palate, local accumulations of intraepithelial nerve fibers which branched and showed terminal enlargements were detected. Intra- and perigemmal nerve fibers of chemosensory corpuscles could be stained. In the palatoglossal arch, numerous elongated papillae were seen containing nerve fiber plexus showing a complicated arborization pattern. In part, the collaterals penetrated the epithelium. The soft palate contained only a small number of lower but broader papillae which were covered by a more expanded intraepithelial nerve fiber plexus. In both regions, anastomoses between the branches of single nerve fibers were sometimes seen. Solitary delicate nerve fiber endings were loosely distributed throughout the epithelia. In addition, intrapapillar nerve endings, which were enclosed by a slightly stained capsule, were intensely stained; they showed characteristic lateral protuberances. The Merkel's discs were visualized as the components of a terminal network of nerve fiber branches. The observed differences in the shape and locations of the nerve terminations suggest different functions of these nerve fibers. Due to the low costs of the staining procedure and its ease in handling, it is well-suited for a mappingof the innervation pattem of the whole oral mucosa
- PublicationOpen AccessNerve distributions in insertional Achilles tendinopathy - a comparison of bone, bursae and tendon(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Andersson, Gustav; Backman, Ludvig J.; Christensen, Jens; Alfredson, HàkanBackground/Aim. In a condition of pain in the Achilles tendon insertion there are multiple structures involved, such as the Achilles tendon itself, the retrocalcaneal bursa and a bony protrusion at the calcaneal tuberosity called Haglund’s deformity. The innervation patterns of these structures are scarcely described, and the subcutaneous calcaneal bursa is traditionally not considered to be involved in the pathology. This study aimed at describing the innervation patterns of the four structures described above to provide a better understanding of possible origins of pain at the Achilles tendon insertion. Methods. Biopsies were taken from 10 patients with insertional Achilles tendinopathy, which had pathological changes in the subcutaneous and retrocalcaneal bursae, a Haglund deformity and Achilles tendon tendinopathy as verified by ultrasound. The biopsies were stained using immunohistochemistry in order to delineate the innervation patterns in the structures involved in insertional Achilles tendinopathy. Results. Immunohistochemical examinations found that the subcutaneous bursa scored the highest using a semi-quantitative evaluation of the degree of innervation when compared to the retrocalcaneal bursa, the Achilles tendon, and the calcaneal bone. Conclusions. These findings suggest that the subcutaneous bursa, which is traditionally not included in surgical treatment, may be a clinically important factor in insertional Achilles tendinopathy.
- PublicationOpen AccessSubstance P and calcitonin gene-related peptide intrinsic choroidal neurons in human choroidal whole-mounts(Murcia : F. Hernández, 2008) Hoz, Ana de; Ramírez, Ana I.; Salazar, Juan J.; Rojas, Blanca; Ramírez, José M.; Triviño, AlbertoTo determine the presence in the human choroid of substance P (SP)-and calcitonin gene-related peptide (CGRP) positive intrinsic choroidal neurons (ICNs), choroidal whole-mounts were processed for indirect immunofluorescence. An antibody to a component of the neuronal cytoskeleton, neurofilament 200 kDa (NF-200), was combined with antibodies to SP and to CGRP (neuropeptides proper to the sensory nervous system). The human choroid possesses numerous SP(+) and CGRP(+) ICNs. These neurons were observed in the suprachoroid, both in isolation and forming microganglia. For both types of ICNs studied, neurons were more numerous in the temporal than in the nasal regions. In both locations, SP(+) and CGRP(+) ICNs were more abundant in the central choroid (the choroid underneath the macular area of the retina), with cell density diminishing outwards to the choroidal periphery. There were no appreciable differences between the two populations of ICNs studied in terms of size, morphology or immunostaining characteristics. In conclusion, given that peripheral sensory innervation could be involved in the regulation of both choroidal blood flow and vascular architecture, the SP(+) and CGRP(+) ICNs described for the first time in the present work may be involved in these mechanisms of vascular regulation.
- PublicationOpen AccessUltrastructural localization of acetylcholinesterase AChE activity in the chicken Harderian gland(Murcia : F. Hernández, 1999) Hiramatsu, K.; Ohshima, K.Localization of acetylcholinesterase (AChE) was investigated in the chicken Harderian gland at the electron microscopic level. Nerve cells in the pterygopalatine ganglion showed AChE activity. They had a pale and large nucleus which was round or oval in shape. Reaction product of AChE was detected between the nuclear envelopes, in the cisterna of rough endoplasmic reticulum and the lumen of the Golgi lamellae, and on the plasma membrane of the nerve cell. In the interstitium of the gland, nerve fibers showing AChE activity were easily found. They were often seen in the perivascular space and between plasma cells. These neve fibers had varicosities in contact with plasma cells and the endothelium or the smooth muscle fiber of the blood vessels. AChE-positive varicosities or terminals contained many small clear vesicles (about 50nm in diameter) and a few large dense-cored vesicles (about 100 nm in diameter). No contacts of nerve fibers with acinar cells or the ductal epithelium were observed in the present study. Our data indicate that cholinergic nerves play distinct roles in the regulation of the immune function of the chicken Harderian gland.