RESUMO
Objective: To investigate the neural connections between Shenmen (HT7)-heart and the brain by observing the tracing viruses co-labeled brain nuclear groups after injection of the pseudorabies viruses (PRV), the reverse transsynaptic virus tracer carrying different fluorescent protein genes, into the myocardium and Shenmen (HT7) point, respectively.Methods: Pseudorabies virus 531 (PRV531) carrying the green fluorescent protein gene and pseudorabies virus 724 (PRV724) carrying the red fluorescent protein gene were injected into the left ventricular wall and Shenmen (HT7) point area of the left forelimb of six C57BL/6 mice, respectively. After 120 h, whole brain tissue was extracted under 4％ paraformaldehyde perfusion to prepare brain sections. Neuronal co-labeling with the tracing viruses was observed under fluorescence microscopy. Results: Co-labeled signals from the mouse ventricular wall and Shenmen (HT7) point region were found at all levels of the mouse central nervous areas, such as the cerebral cortex, hypothalamus, midbrain, pons, and medulla oblongata. The number of co-labeled neurons was higher in the primary motor area, the hypothalamic paraventricular nucleus, the subceruleus nucleus, and the paramedian reticular nucleus. Conclusion: There is a neural connection between Shenmen (HT7), the heart, and the brain, which may be most closely related to the autonomic nervous system.
RESUMO
OBJECTIVE: To collect literatures on the application of neural tracing technique in experimental acupuncture research, and summarize its application status in acupoints, meridians and Zang-fu organs. METHODS: We collected papers published from databases of CNKI (1979-2017), Wan-Fang (1990-2017), VIP (1989-2017) and PubMed(1997-2017)by using keywords of "neural tracing technique" "neuroanatomic tracing" "neural tracers" and "acupuncture" "electroacupuncture" "auricular acupuncture" "eye acupuncture" "meridians" "acupuncture points" "acupoint injection", and made a summary about the current state of application of neural tracing technique in the fields of acupoint, meridian and Zang-fu organs (viscera). RESULTS: A total of 94 articles were collected, the most commonly used neural tracers were horseradish peroxidase and cholera toxin subunit B. The experimental animals used were rat, rabbit, cat, monkey, etc., and injection site was acupoint. After the injection of neural tracers, the survival time of animals was range from 1 day to 12 weeks, and the labelled tissues included neurons and nerve fibers of the sensory, motor, and autonomic systems. The outcomes of neural labeling mainly revealed the segmental pattern, neuroanatomical connection (neural pathways/circuits) and chemical features (shown by immunohistochemical staining) of neurons and nerve fibers innervating both the acupoints and visceral organs, suggesting their involvement in the effect of acupuncture and moxibustion treatment. CONCLUSION: This application of neural tracing technology help us understand the under-lying mechanisms of acupuncture and moxibustion interventions from different perspectives of neural pathways/circuits and related chemical properties, which also lays a greater role for this technology in future experimental acupuncture research.
RESUMO
OBJECTIVE: To explore the neuroanatomical basis of acupoint-visceral correlation by studying the distribution of the neurons associated with acupoint "Shenshu" (BL 23) area and adrenal gland in rats. METHODS: AF 488-CTB and AF 594-CTB were injected into the left side of BL 23 area and adrenal gland in the same rat respectively. Three days after injection, the dorsal root ganglions (DRG), sympathetic chain, and spinal cord were dissected out from the perfused rats. The neuronal labeling with AF 488/594-CTB was directly observed on the sections under a fluorescent microscope or a laser scanning confocal microscope. RESULTS: All neural labeling was observed in the injection side. The sensory neurons associated with both acupoint BL 23 and adrenal gland distributed from thoracic (T) 10 to lumbar (L) 2 DRG with high concentration in T 12-T 13 and T 11-T 12, respectively, in which some of them were simultaneously labeled with both AF 488/594-CTB and located in T 12-L 1 DRG. For the sympathetic innervation, the postganglionic neurons correlated with BL 23 and adrenal gland were labeled with AF 488/594-CTB separately in the sympathetic chain at the lumbar segments, while the labeled preganglionic neurons were only observed at the lateral horn of T 11-T 13 spinal segments in the cases of adrenal gland. In addition, the labeled motor neurons were mainly detected in the spinal ventral horn at cervical (C) 7-C 8 and T 11-L 1 segments. CONCLUSION: These results indicate that there are segmental correlation between BL 23 and adrenal gland on the sensory and sympathetic innervations, and this correlation might be a neural pathway for modulating the function role of adrenal gland through BL 23 needling.
RESUMO
Objective:To investigate the distribution of serotonin 5-HT2A and 5-HT7 receptors in different motoneuron populations in spinal Onuf’s nucleus of male rats,and their relationship with the central modulation of the urogenital functions.Methods:Ten adult SD rats(eight male rats and two female rats)were randomly divided into gender control group(n=4,include two male and two female rats)and retrograde tracing group(male rats,n=6).In gender control group,after the spinal L5~6 segments were removed and cut,sections were treated with 5-HT2A or 5-HT7 receptors immunohistochemical stain.In retrograde tracing group,recombinant pseudorabies virus(PRV-152 )were injected into right ischiocavernosus muscles(IC group;n=3)or right external urethral sphincter(EUS group;n=3),respectively.Four days after injection,the L5~6 segments were removed,cut,and treated with 5-HT2A or 5-HT7 receptors double-labeling immunofluorescence stain.Sections were observed by light microscopy or the laser confocal microscopy to detect the distribution of 5-HT2A、5-HT7 receptors in Onuf’s nucleus.Results:5-HT2A receptors immunoreactivity mainly located in the medial part of the dorsolateral nucleus(DLN)of Onuf’s nucleus,and 5-HT7 receptors mainly located in the lateral part of the DLN;5-HT2A and 5-HT7 receptors expression of male rats in Onuf’s nucleus were more dense than female rats.Immunofluorescence double-labeling detection exhibited that 5-HT2A receptors mainly distributed in surrounding motoneurons innervating the IC muscles,and 5-HT7 receptors mainly expressed in neurons innervating the EUS.Conclusion:5-HT2A and 5-HT7 receptors are heterogeneously distributed in different motoneurons of Onuf’s nucleus innervating different striated muscles.5-HT2A receptor might be preferentially involved in regulation of the sexual reflex,and the 5-HT7 receptor may be mainly participated in the micturition reflex.