Limits...
Identification of Primo-Vascular System in Abdominal Subcutaneous Tissue Layer of Rats.

Lim CJ, Lee SY, Ryu PD - Evid Based Complement Alternat Med (2015)

Bottom Line: The distribution pattern of nodes and vessels in the scPVS closely resembled that of the conception vessel meridian and its acupoints.In conclusion, our results newly revealed that the PVS is present in the abdominal subcutaneous tissue layer and indicate that the scPVS tissues are closely correlated with acupuncture meridians.Our findings will help to characterize the PVS in the other superficial tissues and its physiological roles.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, Republic of Korea.

ABSTRACT
The primo-vascular system (PVS) is a novel network identified in various animal tissues. However, the PVS in subcutaneous tissue has not been well identified. Here, we examined the putative PVS on the surface of abdominal subcutaneous tissue in rats. Hemacolor staining revealed dark blue threadlike structures consisting of nodes and vessels, which were frequently observed bundled with blood vessels. The structure was filled with various immune cells including mast cells and WBCs. In the structure, there were inner spaces (20-60 µm) with low cellularity. Electron microscopy revealed a bundle structure and typical cytology common with the well-established organ surface PVS, which were different from those of the lymphatic vessel. Among several subcutaneous (sc) PVS tissues identified on the rat abdominal space, the most outstanding was the scPVS aligned along the ventral midline. The distribution pattern of nodes and vessels in the scPVS closely resembled that of the conception vessel meridian and its acupoints. In conclusion, our results newly revealed that the PVS is present in the abdominal subcutaneous tissue layer and indicate that the scPVS tissues are closely correlated with acupuncture meridians. Our findings will help to characterize the PVS in the other superficial tissues and its physiological roles.

No MeSH data available.


Related in: MedlinePlus

Identification of the threadlike structures in the rat abdominal subcutaneous tissue using Hemacolor staining. ((a), (b), and (c)) Appearance of the subcutaneous area after serial applications of three Hemacolor solutions: Solution 1 (methanol fixative, (a)), Solution 2 (eosin stain, (b)), and Solution 3 (methylene blue stain, (c)). Dotted line is the abdominal middle line. (d) Appearance of the subcutaneous area after wash-out of Hemacolor solutions. Note the dark blue threadlike structures (arrows). See the Materials and Methods sections for a description of the protocol.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4561983&req=5

fig2: Identification of the threadlike structures in the rat abdominal subcutaneous tissue using Hemacolor staining. ((a), (b), and (c)) Appearance of the subcutaneous area after serial applications of three Hemacolor solutions: Solution 1 (methanol fixative, (a)), Solution 2 (eosin stain, (b)), and Solution 3 (methylene blue stain, (c)). Dotted line is the abdominal middle line. (d) Appearance of the subcutaneous area after wash-out of Hemacolor solutions. Note the dark blue threadlike structures (arrows). See the Materials and Methods sections for a description of the protocol.

Mentions: For the scanning electron microscopy (SEM), scPVS and lymphatic vessels were collected from the rats. The procedure for harvesting the scPVS tissue was the same as that used in the sampling by Hemacolor staining. The harvesting procedure for the lymphatic vessel was as follows: under deep anesthesia, the abdomen of the rat was incised, and the lymphatic vessel was sampled under a stereomicroscope around the kidney by reference to the previous report [21]. The scPVS attached hypodermis layer (Figure 3(a)) was isolated from the area of umbilicus on the rat abdominal middle line (Figures 2(a) and 11(a)). The sampled scPVS and lymphatic vessels were kept in 2% paraformaldehyde for 24 hr and primary fixed by Karnovsky's fixation for 2 hr. After washing them with 0.05 M sodium cacodylate buffer 3 times for a total of 10 min, the samples were postfixed by 2% osmium tetroxide (1 mL) and 0.1 M cacodylate buffer (1 mL) and washed twice using distilled water for 5 s each. Dehydration was conducted using a series of ethanol of 30%, 50%, 70%, 90%, and 100% for 10 min each, and then the samples were dried using a critical point dryer (CPD 030, BAL-TEC) for 1 hr, coated by a sputter coater (EM ACE200, Leica), and observed under a field-emission SEM (SIGMA, Carl Zeiss, UK). For transmission electron microscopy (TEM), several procedures were added to those of SEM. (1) After washing the samples with distilled water, en bloc staining was conducted with 0.5% uranyl acetate for 30 min; (2) after dehydration with ethanols, a transition procedure using propylene oxide was conducted twice for 10 min each; (3) then the samples were embedded into Spurr's resin (2 mL), sectioned, and observed under a TEM (JEM1010, JEOL, Japan).


Identification of Primo-Vascular System in Abdominal Subcutaneous Tissue Layer of Rats.

Lim CJ, Lee SY, Ryu PD - Evid Based Complement Alternat Med (2015)

Identification of the threadlike structures in the rat abdominal subcutaneous tissue using Hemacolor staining. ((a), (b), and (c)) Appearance of the subcutaneous area after serial applications of three Hemacolor solutions: Solution 1 (methanol fixative, (a)), Solution 2 (eosin stain, (b)), and Solution 3 (methylene blue stain, (c)). Dotted line is the abdominal middle line. (d) Appearance of the subcutaneous area after wash-out of Hemacolor solutions. Note the dark blue threadlike structures (arrows). See the Materials and Methods sections for a description of the protocol.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4561983&req=5

fig2: Identification of the threadlike structures in the rat abdominal subcutaneous tissue using Hemacolor staining. ((a), (b), and (c)) Appearance of the subcutaneous area after serial applications of three Hemacolor solutions: Solution 1 (methanol fixative, (a)), Solution 2 (eosin stain, (b)), and Solution 3 (methylene blue stain, (c)). Dotted line is the abdominal middle line. (d) Appearance of the subcutaneous area after wash-out of Hemacolor solutions. Note the dark blue threadlike structures (arrows). See the Materials and Methods sections for a description of the protocol.
Mentions: For the scanning electron microscopy (SEM), scPVS and lymphatic vessels were collected from the rats. The procedure for harvesting the scPVS tissue was the same as that used in the sampling by Hemacolor staining. The harvesting procedure for the lymphatic vessel was as follows: under deep anesthesia, the abdomen of the rat was incised, and the lymphatic vessel was sampled under a stereomicroscope around the kidney by reference to the previous report [21]. The scPVS attached hypodermis layer (Figure 3(a)) was isolated from the area of umbilicus on the rat abdominal middle line (Figures 2(a) and 11(a)). The sampled scPVS and lymphatic vessels were kept in 2% paraformaldehyde for 24 hr and primary fixed by Karnovsky's fixation for 2 hr. After washing them with 0.05 M sodium cacodylate buffer 3 times for a total of 10 min, the samples were postfixed by 2% osmium tetroxide (1 mL) and 0.1 M cacodylate buffer (1 mL) and washed twice using distilled water for 5 s each. Dehydration was conducted using a series of ethanol of 30%, 50%, 70%, 90%, and 100% for 10 min each, and then the samples were dried using a critical point dryer (CPD 030, BAL-TEC) for 1 hr, coated by a sputter coater (EM ACE200, Leica), and observed under a field-emission SEM (SIGMA, Carl Zeiss, UK). For transmission electron microscopy (TEM), several procedures were added to those of SEM. (1) After washing the samples with distilled water, en bloc staining was conducted with 0.5% uranyl acetate for 30 min; (2) after dehydration with ethanols, a transition procedure using propylene oxide was conducted twice for 10 min each; (3) then the samples were embedded into Spurr's resin (2 mL), sectioned, and observed under a TEM (JEM1010, JEOL, Japan).

Bottom Line: The distribution pattern of nodes and vessels in the scPVS closely resembled that of the conception vessel meridian and its acupoints.In conclusion, our results newly revealed that the PVS is present in the abdominal subcutaneous tissue layer and indicate that the scPVS tissues are closely correlated with acupuncture meridians.Our findings will help to characterize the PVS in the other superficial tissues and its physiological roles.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Pharmacology, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, Republic of Korea.

ABSTRACT
The primo-vascular system (PVS) is a novel network identified in various animal tissues. However, the PVS in subcutaneous tissue has not been well identified. Here, we examined the putative PVS on the surface of abdominal subcutaneous tissue in rats. Hemacolor staining revealed dark blue threadlike structures consisting of nodes and vessels, which were frequently observed bundled with blood vessels. The structure was filled with various immune cells including mast cells and WBCs. In the structure, there were inner spaces (20-60 µm) with low cellularity. Electron microscopy revealed a bundle structure and typical cytology common with the well-established organ surface PVS, which were different from those of the lymphatic vessel. Among several subcutaneous (sc) PVS tissues identified on the rat abdominal space, the most outstanding was the scPVS aligned along the ventral midline. The distribution pattern of nodes and vessels in the scPVS closely resembled that of the conception vessel meridian and its acupoints. In conclusion, our results newly revealed that the PVS is present in the abdominal subcutaneous tissue layer and indicate that the scPVS tissues are closely correlated with acupuncture meridians. Our findings will help to characterize the PVS in the other superficial tissues and its physiological roles.

No MeSH data available.


Related in: MedlinePlus