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Neurovascular Primo Bundles at the Kidney Meridian Revealed Using Hemacolor Staining
Innovations in Acupuncture and Medicine volume 8, pages 329–332 (2015)
Neurovascular bundles were observed earlier in connection with the K-meridians of a rat [1]. At that time, the existence of the primo vessel (PV) in the bundle was not noticed despite expectations in accordance with the description of Kim [2]. Applying the hemacolor staining technique, which was recently developed to detect the subcutaneous primo vascular system (PVS) [3], the expected PV was, indeed, revealed. This finding of a neurovascular primo (NVP) bundle is the first example of a PVS found in the peripheral nerves. Until now, the PVS was not regularly observable at specifically defined positions [4]; thus, the current finding is a breakthrough in PVS research in the sense that NVP bundles are repeatedly observed in anatomically well-defined positions. The connection of the PVS with acupuncture was not shown until the recent report on detection of PVS using hemacolor [3]. The NVPs exist at acupuncture meridians such as the putative K and ST meridians. Furthermore, they represent paths from the acupoints in the skin to the brain for either transmission of signals, such as the electric signal of electroacupuncture, or flow of the herbal medicine of pharmacopuncture. This result gives rise to a hope that acupuncture may possibly be used to treat diseases of the brain such as Alzheimer’s and Parkinson’s disease (Fig. 1).
Neurovascular primo bundles. (A) Schematic illustration showing the distribution of the NVP bundles along the putative kidney meridian. The NVP bundles are found near the acupuncture points (K17–K21). The midline shows the CV. (B) The NVP bundles (arrows) in the hypodermis along the kidney meridians of a rat are displayed as a mosaic of stereomicroscope images. The left and the right groups of five NVP bundles (those on one side are indicated by arrows) on the kidney meridians formed two lines that were parallel to the white middle line (linea alba). The left and the right NVP bundles were located about 3–4 mm away from the middle line. (C) Stereomicroscope image of an NVP bundle located in the hypodermis. It is stretched in order to show the whole shape. The upper part entered the dermis layer, and the lower part entered the abdominal muscle layer. The NVP bundle was studied earlier in order to search for the PV along the blood vessels, but no PV was seen. Only a nerve was seen to run along the blood vessels [1]. (D) After staining with hemacolor, the PV (arrow) in the nerve of the NVP bundle emerged as a blue, threadlike structure running along the blood vessels (arrow heads). The existence of the PV was expected from Kim’s [2] description, but was not noticed until the recently developed hemacolor staining method [3] was applied. (E) A piece of a harvested NVP sample. The hemacolor staining showed the parallel curves of the PVs (arrows) in the nerve. The stained parts can be clearly discerned at a higher magnification with a phase contrast microscope in vitro, as shown in (F) and (G). (F) Magnified (400×) views of an NVP bundle; the view was obtained using a phase contrast microscope. The PVs (arrows) were stained with hemacolor. Some mast cells are indicated by arrowheads. (G) The yellow lines are PVs illustrated by Kim [2]. The PVs (arrows) are located at the perineurium and the endoneuria. The artery and vein are indicated by red and blue arrows, respectively. (H) A phase contrast microscope image of the cross-section of an NVP bundle in Optimal Cutting Temperature (OCT) before defreezing. The blue stained spots (arrows) exhibit the positions of the PVs. Most of them are just inside the perineurium, but some are in the fascicle (yellow double-headed arrow). Some blood vessels are indicated by double-headed arrows. (I) After tissue processing and H&E staining, the blue stained spots that appeared in Fig. 1H, implying the positions of the PVs (arrows) just outside of the perineurium, remained clearly, however, there appeared more stained spots either inside or outside of the nerve bundle. This was due to insufficient washing of the hemacolor. Three-dimensional digital histopathology will be performed to show more explicitly what the destiny of the threadlike structure of the PV would be in a three-dimensional way. Two blood vessels are indicated with double-headed arrows. After 4′, 6-Diamidino-2-phenylindole (DAPI) staining of nuclei and washing, the hemacolor disappeared, and (J) the bright mode shows fiber-like structures (arrows) of the nerve with the PV, whereas (K) the fluorescent mode revealed the putative endothelial nuclei (blue rod-shaped structures indicated with arrows) of the PV. The aligned distribution of rod-shaped nuclei (length 10–20 µm) is the hallmark of a PV [2–4]. (L) The NVP is located in the HD layer under the D and penetrates the F to the M. The peripheral nerve goes to the thoracic spinal cord through the ganglion. The insets are (F) and (G), and indicate their respective locations in the peripheral nerve. The nervous PV from the kidney acupoint can provide a path for electric signal transmission from the acupoint to the spine. It can also be a conduit for the herbal medicine that is injected into the acupoint. This path further proceeds to various parts of the brain. Therefore, acupuncture can influence the brain either by electric signal transmissions or by herbal medicine delivery. CV = conception vessel; D = dermis; F = deep fascia; HD = hypodermis; H&E = hematoxylin and eosin; M = abdominal muscle; NVP = neurovascular primo; PV = primo vessel.
References
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Acknowledgements
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (Grant numbers: 2013 R1A1A2011526 and 2013R1A1A2008343).
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Bae, KH., Gil, HJ., Yoo, YY. et al. Neurovascular Primo Bundles at the Kidney Meridian Revealed Using Hemacolor Staining. Innov. Acupunct. Med. 8, 329–332 (2015). https://doiorg.publicaciones.saludcastillayleon.es/10.1016/j.jams.2015.10.002
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DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1016/j.jams.2015.10.002