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1.
Histochem Cell Biol ; 151(2): 161-173, 2019 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-30244428

RESUMEN

The morphological characteristics of baroreceptors in the rat carotid sinus were reevaluated by whole-mount preparations with immunohistochemistry for P2X3 purinoceptors using confocal scanning laser microscopy. Immunoreactive nerve endings for P2X3 were distributed in the internal carotid artery proximal to the carotid bifurcation, particularly in the region opposite the carotid body. Some pre-terminal axons in nerve endings were ensheathed by myelin sheaths immunoreactive for myelin basic protein. Pre-terminal axons ramified into several branches that extended two-dimensionally in every direction. The axon terminals of P2X3-immunoreactive nerve endings were flat and leaf-like in shape, and extended hederiform- or knob-like protrusions in the adventitial layer. Some axons and axon terminals with P2X3 immunoreactivity were also immunoreactive for P2X2, and axon terminals were closely surrounded by terminal Schwann cells with S100 or S100B immunoreactivity. These results revealed the detailed morphology of P2X3-immunoreactive nerve endings and suggested that these endings respond to a mechanical deformation of the carotid sinus wall with their flat leaf-like terminals.


Asunto(s)
Seno Carotídeo/química , Presorreceptores/química , Receptores Purinérgicos P2X3/análisis , Animales , Seno Carotídeo/metabolismo , Inmunohistoquímica , Masculino , Presorreceptores/metabolismo , Ratas , Ratas Wistar , Receptores Purinérgicos P2X3/metabolismo
2.
Int J Mol Sci ; 20(13)2019 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-31269659

RESUMEN

Chronic inflammatory pain is one of the most common complaints that seriously affects patients' quality of life. Previous studies have demonstrated that the analgesic effect of electroacupuncture (EA) stimulation on inflammatory pain is related to its frequency. In this study, we focused on whether the analgesic effects of EA are related to the period of stimulation. Purinergic receptor P2X3 (P2X3) is involved in the pathological process underlying chronic inflammatory pain and neuropathic pain. We hypothesized that 100 Hz EA stimulation alleviated Freund's complete adjuvant (CFA) induced inflammatory pain via regulating P2X3 expression in the dorsal root ganglion (DRG) and/or spinal cord dorsal horn (SCDH). We also assumed that the analgesic effect of EA might be related to the period of stimulation. We found that both short-term (three day) and long-term (14 day) 100 Hz EA stimulation effectively increased the paw withdrawal threshold (PWT) and reversed the elevation of P2X3 in the DRG and SCDH of CFA rats. However, the analgesic effects of 100 Hz EA were not dependent on the period of stimulation. Moreover, P2X3 inhibition or activation may contribute to or attenuate the analgesic effects of 100 Hz EA on CFA-induced inflammatory pain. This result indicated that EA reduced pain hypersensitivity through P2X3 modulation.


Asunto(s)
Electroacupuntura/métodos , Adyuvante de Freund , Manejo del Dolor/métodos , Dolor/inducido químicamente , Receptores Purinérgicos P2X3/análisis , Animales , Ganglios Espinales/patología , Inflamación/inducido químicamente , Inflamación/patología , Inflamación/terapia , Masculino , Dolor/patología , Ratas Sprague-Dawley , Asta Dorsal de la Médula Espinal/patología
3.
Histochem Cell Biol ; 145(2): 131-46, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26475709

RESUMEN

The morphological characteristics of P2X3-immunoreactive nerve endings in the laryngeal mucosa were herein examined using immunohistochemistry with confocal laser microscopy. Ramified intraepithelial nerve endings immunoreactive to P2X3 were distributed in the epiglottis and arytenoid region. The axon terminals of P2X3-immunoreactive ramified endings were beaded or flat in shape. These endings were also immunoreactive to P2X2 and not identical to the nerve endings immunoreactive to Na(+)-K(+)-ATPase α3-subunit, substance P (SP), and calcitonin gene-related peptide (CGRP). P2X3-immunoreactive axon terminals were also immunoreactive to vGLUT1, vGLUT2, and vGLUT3. In addition to ramified endings, P2X3-immunoreactive nerve endings were associated with α-gustducin-immunoreactive solitary chemosensory cells and/or SNAP25-immunoreactive neuroendocrine cells. Furthermore, P2X3-immunoreactive nerve endings were also observed in the taste bud-like chemosensory cell clusters of the stratified squamous epithelium covering epiglottic and arytenoid cartilage. The P2X3-immunoreactive nerve endings that associated with sensory and/or endocrine cells and chemosensory cell clusters were also immunoreactive to P2X2, vGLUT1, vGLUT2, and vGLUT3, but not to SP or CGRP. In conclusion, P2X3-immunoreactive nerve endings may be classified into two types, i.e., intraepithelial ramified nerve endings and nerve endings associated with chemosensory cells and neuroendocrine cells.


Asunto(s)
Mucosa Laríngea/citología , Terminaciones Nerviosas/química , Receptores Purinérgicos P2X3/análisis , Animales , Inmunohistoquímica , Mucosa Laríngea/química , Mucosa Laríngea/inmunología , Rayos Láser , Microscopía Confocal , Terminaciones Nerviosas/clasificación , Terminaciones Nerviosas/inmunología , Ratas , Receptores Purinérgicos P2X3/inmunología
4.
Histochem Cell Biol ; 146(4): 479-88, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27368183

RESUMEN

We investigated the three-dimensional architectures of P2X2-/P2X3-immunoreactive nerve terminals in the rat carotid body using immunohistochemistry with confocal laser microscopy. Nerve endings immunoreactive for P2X2 and P2X3 were associated with clusters of type I cells, whereas some nerve endings were sparsely distributed in a few clusters. Most nerve endings surrounding type I cells were hederiform in shape and extended several flattened axon terminals, which were polygonal or pleomorphic in shape and contained P2X2-/P2X3-immunoreactive products. Three-dimensional reconstruction views revealed that some flattened nerve endings with P2X3 immunoreactivity formed arborized, sac- or goblet-like terminal structures and were attached to type I cells immunoreactive for tyrosine hydroxylase (TH). However, P2X3-immunoreactive axon terminals were sparsely distributed in type I cells immunoreactive for dopamine beta-hydroxylase. Multi-immunolabeling for P2X2, S100, and TH revealed that P2X2-immunoreactive axon terminals were attached to TH-immunoreactive type I cells on the inside of type II cells with S100 immunoreactivity. These results revealed the detailed morphology of P2X2-/P2X3-immunoreactive nerve terminals and suggest that sensory nerve endings may integrate chemosensory signals from clustered type I cells with their variform nerve terminals.


Asunto(s)
Cuerpo Carotídeo/anatomía & histología , Cuerpo Carotídeo/metabolismo , Microscopía Confocal , Terminaciones Nerviosas/metabolismo , Receptores Purinérgicos P2X2/inmunología , Receptores Purinérgicos P2X3/inmunología , Animales , Cuerpo Carotídeo/inmunología , Inmunohistoquímica , Masculino , Terminaciones Nerviosas/inmunología , Ratas , Ratas Wistar , Receptores Purinérgicos P2X2/análisis , Receptores Purinérgicos P2X3/análisis
5.
BJU Int ; 116(2): 293-301, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25430615

RESUMEN

OBJECTIVES: To evaluate whether P2X3 receptors (P2X3R) are expressed in the bladder urothelium and to determine their possible function in modulating purinergic detrusor contractions in the rat urinary bladder. MATERIALS AND METHODS: The expression of urothelial receptors was determined using conventional immunohistochemistry in bladders from normal Sprague-Dawley rats. The urothelial layer was removed by incubation with protamine, and disruption of the urothelium was confirmed using haematoxylin and eosin staining on bladder sections. Open cystometry was used to determine the effects of both urothelial removal as well as intravesical application of a specific P2X3R antagonist on bladder properties from intact and protamine-treated rats. Isometric contractile responses to potassium chloride (KCl) depolarization, electrical field stimulation (EFS) or chemical P2X activation were determined in normal and urothelium-denuded bladder strips, with and without application of the P2X3R antagonist. RESULTS: Immunohistochemical staining showed high expression of P2X3R in the medial and basal layers of the urothelium. Removal of the urothelial layer disturbed normal bladder performance in vivo and eliminated the effects of the P2X3R antagonist on increasing the contractile interval and reducing the amplitude of voiding contractions. Removal of the urothelium did not affect bladder strip contractile responses to KCl depolarization or EFS. Pharmacological inhibition of P2X3R prevented desensitization to P2X-mediated detrusor muscle contractions during EFS only in the strips with an intact urothelium. A concentration-dependent, specific inhibition of P2X3R also prevented desensitization of purinergic contractile responses in intact bladder strips. CONCLUSIONS: In the rat bladder, medial and basal urothelial cells express P2X3R, and specific inhibition of the receptor leads to a more hyporeflexive bladder condition. This pathway may involve P2X3R driving a paracrine amplification of ATP released from umbrella cells to increase afferent transmission in the sub-urothelial sensory plexus and desensitization of P2X1-mediated purinergic detrusor contractions.


Asunto(s)
Contracción Muscular/efectos de los fármacos , Antagonistas del Receptor Purinérgico P2X/farmacología , Receptores Purinérgicos P2X3/metabolismo , Urotelio/efectos de los fármacos , Urotelio/metabolismo , Animales , Femenino , Inmunohistoquímica , Éteres Fenílicos/farmacología , Pirimidinas/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Purinérgicos P2X3/análisis , Vejiga Urinaria/metabolismo , Vejiga Urinaria/fisiología , Urotelio/química
6.
J Comp Neurol ; 529(10): 2771-2788, 2021 07 01.
Artículo en Inglés | MEDLINE | ID: mdl-33550628

RESUMEN

Peripheral sensory neurons are characterized by their size, molecular profiles, and physiological responses to specific stimuli. In mouse, the peptidergic and non-peptidergic subsets of nociceptors are distinct and innervate different lamina of the spinal dorsal horn. The unique molecular signature and neuroanatomical organization of these neurons supports a labeled line theory for certain types of nociceptive stimuli. However, long-standing evidence supports the polymodal nature of nociceptors in many species. We have recently shown that the peptidergic marker, CGRP, and the non-peptidergic marker, P2X3R, show largely overlapping expression at the mRNA level in human dorsal root ganglion (DRG). Herein, our aim was to assess the protein distribution of nociceptor markers, including their central projections, in the human DRG and spinal cord. Using DRGs obtained from organ donors, we observed that CGRP and P2X3R were co-expressed by approximately 33% of human DRG neurons and TrpV1 was expressed in ~60% of human DRG neurons. In the dorsal spinal cord, CGRP, P2X3R, TrpV1, and Nav1.7 proteins stained the entirety of lamina 1-2, with only P2XR3 showing a gradient of expression. This was confirmed by measuring the size of the substantia gelatinosa using Hematoxylin and Eosin staining of adjacent sections. Our findings are consistent with the known polymodal nature of most primate nociceptors and indicate that the central projection patterns of nociceptors are different between mice and humans. Elucidating how human nociceptors connect to subsets of dorsal horn neurons will be important for understanding the physiological consequences of these species differences.


Asunto(s)
Péptido Relacionado con Gen de Calcitonina/análisis , Ganglios Espinales/metabolismo , Nociceptores/metabolismo , Receptores Purinérgicos P2X3/análisis , Asta Dorsal de la Médula Espinal/metabolismo , Adulto , Péptido Relacionado con Gen de Calcitonina/biosíntesis , Femenino , Humanos , Masculino , Persona de Mediana Edad , Receptores Purinérgicos P2X3/biosíntesis
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