• Title/Summary/Keyword: WGA-HRP

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Experimental Study for Innervation of Scalp and Face with WGA-HRP Method (WGA-HRP법을 이용한 두피와 안면부의 신경지배에 관한 연구)

  • Kang, Jun-Goo
    • The Korean Journal of Pain
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    • v.7 no.2
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    • pp.238-241
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    • 1994
  • Aim of this study was to discover the projection area of the first cervical spinal nerve. Subcutaneous injection of wheat germ agglutinin-horseradish peroxidase(WGA-HRP) was done at five points of young dogs scalp and face. After two days of survival time, animals were sacrificed by perfusion through the left ventricle of the heart. Trigeminal ganglion, first and second cervical dorsal root ganglion, superior cervical ganglion, middle cervical ganglion and stellate ganglion were removed. Projection area of wheat germ agglutinin-horseradish peroxidase in vestigated into above ganglions. Projection into the first cervical dorsal root ganglion and stellate ganglion was not found. This experiment is deemed valuable for the study of neuronal connection on the central nervous system.

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Neuroanatomical Studies on the Acupoints Related to the Large Intestine (대장(大腸)과 관련(關聯)된 경혈(經穴)들의 신경해부학적(神經解剖學的) 연구(硏究))

  • Kang, Chang-Soo;Lee, sang-ryoung;Lee, Chang-Hyun;Nam, Yong-Jae;Lee, Kwang-Gyu
    • Journal of Acupuncture Research
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    • v.17 no.2
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    • pp.95-117
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    • 2000
  • The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.

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Neural pathway innervating ductus Deferens of rats by pseudorabies virus and WGA-HRP (흰쥐에서 WGA-HRP와 pseudorabies virus를 이용한 정관의 신경로에 대한 연구)

  • Lee, Chang-Hyun;Chung, Ok-Bong;Ko, Byung-Moon;Lee, Bong-Hee;Kim, Soo-Myung;Kim, In-Shik;Yang, Hong-Hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.11-24
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    • 2003
  • This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

Hypothalamic Orexin-A Projections to Midline Thalamic Nuclei in the Rat

  • Lee, Hyun-S.
    • Animal cells and systems
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    • v.9 no.3
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    • pp.145-152
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    • 2005
  • A retrograde tracer, WGA-apo-HRP-gold, was injected into midline thalamic nuclei and subsequently orexin-A immunostaining was performed on the tuberal region of the hypothalamus in order to investigate orexinergic projections to the midline thalamus. Injection site was targeted within one specific region, i.e., paraventricular, centromedian, rhomboid, reuniens, or intermediodorsal nucleus, but it proved to be either one or a combination of these thalamic nuclei. The distribution of WG/orexin-double-labeled neurons exhibited a general pattern in that the majority of labeled cells were observed within the ventral portion of the lateral hypothalamus as well as the perifornical nucleus (PeF). A small number of double-labeled cells were also observed at the dorsomedial nucleus, the area dorsal to the PeF, dorsal portion of the lateral hypothalamus, and the posterior hypothalamus. These orexin-immunoreactive neurons might have wake-related influences over a variety of functions related with midline thalamic nuclei, which include autonomic control, associative cortical functions, and limbic regulation.

Localization of Sensory Neurons Innervating the Rat Intestine Using the Cholera Toxin B Subunit(CTB) and Wheat Germ Agglutinin-Horseradish Peroxidase(WGA-HRP) (표지방식을 이용한 흰 쥐 복강 내장을 지배하는 감각신경세포체와 신경섬유의 표지부위)

  • Lee, Dong-Hyup;Lee, Chang-Hyun;Lee, Moo-Sam
    • Journal of Yeungnam Medical Science
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    • v.15 no.1
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    • pp.75-96
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    • 1998
  • The local arrangement of sensory nerve cell bodies and nerve fibers in the brain stem, spinal ganglia and nodose ganglia were observed following injection of cholera toxin B subunit(CTB) and wheat germ agglutinin-horseradish peroxidase(WGA-HRP) into the rat intestine. The tracers were injected in the stomach(anterior and posterior portion), duodenum, jejunum, ileum, cecum, ascending colon or descending colon. After survival times of 48-96 hours, the rats were perfused and their brain, spinal and nodose ganglia were frozen sectioned ($40{\mu}m$). These sectiones were stained by CTB immunohistochemical and HRP histochemical staining methods and observed by dark and light microscopy. The results were as follows: 1. WGA-HRP labeled afferent terminal fields in the brain stem were seen in the stomach and cecum, and CTB labeled afferent terminal fields in the brain stem were seen in all parts of the intestine. 2. Afferent terminal fields innervating the intestine were heavily labeled bilaterally gelalinous part of nucleus of tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS(comNTS), medial part of NTS(medNTS), wall of the fourth ventricle, ventral border of area postrema and comNTS in midline dorsal to the central canal. 3. WGA-HRP labeled sensory neurons were observed bilaterally within the spinal ganglia, and labeled sensory neurons innervating the stomach were observed in spinal ganglia $T_2-L_1$ and the most numerous in spinal ganglia $T_{8-9}$. 4. Labeled sensory neurons innervating the duodenum were observed in spinal ganglia $T_6-L_2$ and labeled cell number were fewer than the other parts of the intestines. 5. Labeled sensory neurons innervating the jejunum were observed in spinal ganglia $T_6-L_2$ and the most numerous area in the spinal ganglia were $T_{12}$ in left and $T_{13}$ in right. 6. Labeled sensory neurons innervating the ileum were observed in spinal ganglia $T_6-L_2$ and the most numerous area in the spinal ganglia were $T_{11}$ in left and $L_1$ in right. 7. Labeled sensory neurons innervating the cecum were observed in spinal ganglia $T_7-L_2$ and the most numerous area in the spinal ganglia were $T_{11}$ in left and $T_{11-12}$ in right. 8. Labeled sensory neurons innervating the ascending colon were observed in spinal ganglia $T_7-L_2$ in left, and $T_9-L_4$ in right. The most numerous area in the spinal ganglia were $T_9$ in left and $T_{11}$ in right. 9. Labeled sensory neurons innervating the descending colon were observed in spinal ganglia $T_9-L_2$ in left, and $T_6-L_2$ in right. The most numerous area in the spinal ganglia were $T_{13}$ in left and $L_1$ in right. 10. WGA-HRP labeled sensory neurons were observed bilaterally within the nodose ganglia, and the most numerous labeled sensory neurons innervating the abdominal organs were observed in the stomach. 11. The number of labeled sensory neurons within the nodose ganglia innervating small and large intestines were fewer than that of labeled sensory neurons innervating stomach These results indicated that area of sensory neurons innervated all parts of intestines were bilaterally gelatinous part of nucleus tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS (comNTS), medial part of NTS, wall of the fourth ventricle, ventral border of area postrema and com NTS in midline dorsal to the central canal within brain stem, spinal ganglia $T_2-L_4$ and nodose ganglia. Labeled sensory neurons innervating the intestines except the stomach were observed in spinal ganglia $T_6-L_4$. The most labeled sensory neurons from the small intestine to large intestine came from middle thoracic spinal ganglia to upper lumbar spinal ganglia.

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Localization of Motor and Sensory Neurons Innervating Kidney, Shinsu(BL23) and Kyongmun(GB25) in the Rat (흰쥐의 신장(腎臟) 신유(腎兪) 경문(京門)을 지배하는 운동(運動)과 감각신경세포체(感覺神經細胞體)에 대한 연구(硏究))

  • Ryu, Suk-Hyun;Lee, Chang-Hyeon;Lee, Sang-Ryong
    • The Journal of Korean Medicine
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    • v.18 no.1
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    • pp.385-398
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    • 1997
  • The location and local arrangement of motor, sensory neurons within brain stem, nodose ganglia, spinal ganglia and sympathetic ganglia projecting to rat's kidney and meridian point BL 23, GB 25 were investigated by HRP immunohistochemical methods following injection of 5% WGA-HRP into left kidney and meridian point BL 23, GB 25. Following injection of WGA-HRP into left kidney, anterogradely labelled sensory neurons were founded within either nodose ganglia and spinal ganglia. The sensory neurons innervating rat's left kidney were observed within spinal ganglia $T_{7}{\sim}L_3$. Sympathetic motor neurons innervating rat's left kidney were labelled within left suprarenal ganglia, either celiac ganglia, superior mesenteric ganglia, and sympathetic chain ganglia $T_{1}{\sim}L_3$. Sympathetic chain ganglia were concentrated in $T_{12}{\sim}L_1$. The sensory neurons innervating rat's meridian point BL 23 were founded within spinal ganglia $T_{2}{\sim}L_2$. They were numerous in spinal in ganglia $T_{10}{\sim}T_{12}$. Sympathetic motor neurons innervating rat's meridian point BL 23 were observed in suprarenal ganglia and greater splanchnic trunk, sympathetic chain ganglia from $T_1$ to $L_3$. They were concentrated in $T_{12}{\sim}L_3$. The sensory neurons innervating rat's meridian point GB 25 were labelled within spinal ganglia $T_{6}{\sim}T_{13}$. They were numerous in from T10 to $T_{12}$. Sympathetic motor neurons innervating rat's meridian point GB 25 were labelled within greater splanchnic trunk and sympathetic chain ganglia $T_{12}{\sim}L_3$. They were concentrated in $T_{13}{\sim}L_1$. This results neuroanatomically imply that the location of rat's motor and sensory neurons innervating meridian point BL 23 and GB 25 were closely related that of innervating kidney.

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Central Projections of Sensory Information Produced by Topical Application of Capsaicin to the Tongue in the Cat

  • Hong, Seung-Kil;Nahm, Sook-Hyun;Han, Hee-Chul;Suh, Young-Suk;Won, Ran;Lee, Bae-Hwan
    • The Korean Journal of Physiology
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    • v.25 no.1
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    • pp.87-99
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    • 1991
  • In order to elucidate whether capsaicin applied topically acts as a pain-producing subastance or as a tastant, neuronal activities of the chorda tympani nerve(CN), lingual nerve(LN), solitary tract nucleus(STN), and trigeminal nucleus(TGN) were recorded while thermal and taste stimuli, and capsaicin were being applied topically, and algesics intra-arterially to the tongue of cats anesthetized with ${\alpha}-chloralsoe$. In addition, the STN neurons were examined after wheat germ agglutinin-horseradish peroxidase(WGA-HRP) was applied to the CN. The CN fibers responded to taste and thermal stimuli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to taste and thermal stimuli. The LN fibers mainly responded to mechanical and thermal stimuli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to algesics. The STN neurons responded to taste and thermal stiumli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to taste and thermal stiumli in somewhat different fashion from those of the CN fibers. The TGN neurons mainly responded to mechanical stimuli, algesics, and capsaicin. Correlations between responses to capsaicin and any others were not significant. After WGA-HRP was applied to the CN, the STN neurons which receive input from the CN were identified largely in the medio-ventral portion to the solitary tract. These results suggest that capsaicin produce taste as well as pain sensation. Sensory information evoked by capsaicin can be conveyed to the STN, especially medio-ventral portion, via the CN as gustatory information on the one hand, and to the STN or TGN via the LN as noxious information on the other. In addiation, the noxious information may be conveyed to the STN via the CN.

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The Projection from the Lateral Reticular Nucleus to the Cerebellar Vermal Lobule VI in the Rat: A Retrograde Labelling Study Using Horseradish Peroxidase (쥐의 외측 망상핵으로부터 소뇌충부 제6엽 내의 각 소엽으로 신경 경로에 관한 연구)

  • 이현숙
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.26-35
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    • 1996
  • The projection from the lateral reticular nucleus (LRN) to three subdivisions of the cerebellar vermal lobule VI was studied in the rat by utilizing the retrograde transport of wheatgerm agglutinin-conjugated horseradish peroxidase. Labelled neurons were located bilaterally throughout the LRN, but with ipsilateral predominance. There seemed to be a dorsal4o-ventral transition in the ipsilateral magnocellular neurons projecting to the cerebellar vermal lobules VIa-to-Vic. In the contralateral side, cells projecting to vermal lobule Via were observed at more rostral sections, whereas those projecting to lobule Vic were located at caudal sections. There were relatively few labelled neurons in parvocellular or subtrigeminal divisions at either side. Computer-aided three dimensional reconstruction of LRN projections to lobules Via/Vib or lobules Vib/Vic exhibited extensive oberlap within each combination of injection cases.

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Morphological Studies on the Localization of Neurons Projecting to the Meridian Points Related to the Facial Nerve Paralysis in the Rat Using the Neural Tracers (신경추적자(神經追跡子)를 이용한 얼굴신경마비(神經痲痺)와 관련(關聯)된 혈(穴)들을 지배(支配)하는 신경세포체(神經細胞體)의 표식부위(標識部位)에 대(對)한 형태학적(形態學的) 연구(硏究))

  • Kim, Jum-Young;Lee, Sang-Ryoung;Lee, Chang-Hyun
    • The Journal of Korean Medicine
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    • v.18 no.1
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    • pp.58-71
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    • 1997
  • In order to the location and local arrangement of nerve cell bodies and nerve fibers projecting to the meridian points related to facial nerve paralysis in the rat using the neural tracers, CTB and WGA-HRP, labeled neurons the were investigated by immunohistochemical and HRP histochemical methods following injection of 2.5% WGA-HRP and 1% CTB into Hyopko$(S_6)$. Chichang$(S_4)$, Sugu$(GV_{26})$, Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$. Following injection of Hyopko$(S_6)$, Chichang$(S_4)$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus, reticular nucleus and hypoglossal nucleus. labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, sensory root of trigeminal nerve, oral, interpolar and caudal part of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus and $C_{1-2}$ spinal ganglia. Following injection of Sugu$(GV_{26})$, labeled motor neurons were founded in facial nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in spinal trigeminal tract, trigeminal motor nucleus, mesencephalic trigeminal tract, oral. interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, lateral reticular nucleus, dorsal part of reticular part and $C_{1-2}$ spinal ganglia. Following injection of Sajukkong$(TE_{23})$ and Yangbaek$(G_{14})$, labeled motor neurons were founded in facial nucleus, trigeminal motor nucleus. Labeled sensory neurons were founded in trigeminal ganglia and $C_{1-2}$ spinal ganglia. sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius, inferior olovary nucleus, medullary reticular field and lamina I-IV of $C_{1-2}$ spinal cord. Location of nerve cell body and nerve fibers projecting to the meridian points related to the facial nerve paralysis in the rats were found in facial nucleus and trigeminal motor nucleus. Sensory neurone were found in trigeminal ganglia and $C_{1-2}$ spinal ganglia. Sympathetic motor neurons were found in superior cervical ganglia. Sensory fibers labeled in brainstem were found in mesencephalic trigeminal tract, oral, interpolar and caudal parts of trigeminal nucleus, area postrema, nucleus tractus solitarius. lateral reticular nucleus, medullary reticular field.

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MORPHOLOGICAL ANALYSIS OF DEVELOPMENTAL CHANGES IN SOMA AREA OF DIGASTRIC MOTONEURONS IN THE RAT TRIGEMINAL MOTOR NUCLEI (흰쥐의 두힘살근 운동신경세포의 발달과정에 관한 형태학적 분석)

  • Kim, Jae-Hyun;Park, Mi-Hwa;Paik, Sang-Kyoo;Ma, Su-Kyung;Baek, Sang-Heum;Cha, Du-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.2
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    • pp.137-142
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    • 2005
  • To analyze the developmental changes in soma diameters of digastric motoneurons, wheat-germ agglutinin conjugated horseradish peroxidase (WGA-HRP) was injected into the digastric muscle and visualized the retrogradely HRP-labeled motoneurons through tungstate/tetramethylbenzidine (TMB) and following diaminobenzidine (DAB) reactions. The results obtained from Sprague-Dawley rats at postnatal days 1 (P1), 10 (P10) and 30 (P30) indicated as follows: firstly, soma diameters of digastric motoneurons showed unimodal distribution in all postnatal days examined; secondly, the period of P1 to P10 (period 1) showed about 2 times faster growth rate than that of P10 to P30 (period 2); thirdly, the smallest soma examined in each postnatal day exhibited slower growth rate with that of the largest one (increase ratio in soma diameters from P1 to P30, smallest vs. largest = 1.62 : 1.93); Finally, relative growth rates a day showed again that period 1 had faster growth rate than that of period 2. Consequently, developmental changes in soma diameters of digastric motoneurons resulted in very different growth rates between both periods. This implies that the growth of the soma is almost completing within P10 and thereafter growing slowly. The period 1 and 2 are corresponding to sucking and sucking/masticatory period, respectively. Therefore present study providing morphological changes in soma diameters of digastric motoneurons suggests that both periods and their different growth rates of the motoneurons in each period may closely be related with each other.