• Title/Summary/Keyword: microtome

Search Result 45, Processing Time 0.019 seconds

Immunocytochemical Localization of Parvalbumin and Calbindin-D 28K in Monkey Dorsal Lateral Geniculate Nucleus (원숭이 외측슬상체배측핵에서 칼슘결합단백 Parvalbumin과 Calbindin-D 28K의 분포)

  • Ko, Seung-Hee;Bae, Choon-Sang;Park, Sung-Sik
    • Applied Microscopy
    • /
    • v.24 no.4
    • /
    • pp.61-77
    • /
    • 1994
  • The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.

  • PDF

Neuroprotective Effects of Modified Yuldahanso-tang (MYH) in a Parkinson's Disease Mouse Model (MPTP로 유도된 Parkinson's disease 동물 모델에서 열다한소탕 가감방 (MYH)의 신경 세포 보호 효과)

  • Go, Ga-Yeon;Kim, Yoon-Ha;Ahn, Taek-Won
    • Journal of Sasang Constitutional Medicine
    • /
    • v.27 no.2
    • /
    • pp.270-287
    • /
    • 2015
  • Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

CROSS-SECTIONAL MORPHOLOGY AND MINIMUM CANAL WALL WIDTHS IN C-SHAPED ROOT OF MANDIBULAR MOLARS (C-shaped canal의 절단면 분석을 통한 근관형태의 변화와 근관과 치아외벽간의 최소거리 분석에 관한 연구)

  • Song, Byung-Chul;Cho, Yong-Bum
    • Restorative Dentistry and Endodontics
    • /
    • v.32 no.1
    • /
    • pp.37-46
    • /
    • 2007
  • The C-shaped canal system is an anatomical variation mostly seen in mandibular second molars, although it can also occur in maxillary and other mandibular molars. The main anatomical feature of C-shaped canals is the presence of fins or web connecting the individual root canals. The complexity of C-shaped canals prevents these canals from being cleaned, shaped, and obturated effectively during root canal therapy, and sometimes it leads to an iatrogenic perforation from the extravagant preparation. The purpose of this study was to provide further knowledge of the anatomical configuration and the minimal thickness of dentinal wall according to the level of the root. Thirty extracted mandibular second molars with fused roots and longitudinal grooves on lingual or buccal surface of the root were collected from a native Korean population. The photo images and radiographs from buccal, lingual, apical direction were taken. After access cavity was prepared, teeth were placed in 5.25% sodium hypochlorite solution for 2 hours to dissolve the organic tissue of the root surface and from the root canal system. After bench dried and all the teeth were embedded in a self-curing resin. Each block was sectioned using a microtome (Accutom-50, Struers, Denmark) at interval of 1 mm. The sectioned surface photograph was taken using a digital camera (Coolpix 995, Nikon, Japan) connected to the microscope. 197 images were evaluated for canal configurations and the minimal thickness of dentinal wall between canal and external wall using 'Root Thickness Gauge Program' designed with Visual Basic. The results were as follows : 1. At the orifice level of all teeth, the most frequent observed configuration was Melton's Type C I (73%), however the patterns were changed to type C II and C III when the sections were observed at the apical third. On the other hand, the type C III was observed at the orifice level of only 2 teeth but this type could be seen at apical region of the rest of the teeth. 2. The C-shaped canal showed continuous and semi-colon shape at the orifice level, but at the apical portion of the canal there was high possibility of having 2 or 3 canals 3. Lingual wall was thinner than buccal wall at coronal, middle, apical thirds of root but there was no statistical differences.

The Effect of Selenium and Vitamin E on Activity of Enzyme Related to the Lipid Peroxidation in Rat with Alcohol Administration (식이내 Selenium과 Vitamin E가 Alcohol을 섭취한 흰쥐의 간 지질 과산화에 관련된 효소의 활성에 미치는 영향)

  • 김갑순;정승용;김석환
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.22 no.2
    • /
    • pp.116-126
    • /
    • 1993
  • The purposes of this study were to investigate the effect of seleniumc (Se) and vitamin E on activity of enzyme relevant to lipid peroxidation in alcohol administrated rats. Seventy two male rats of Sprague-Dawley strain weighing about 58~62g were divided into 12groups. The dietary Se levels were 0, 0.4 and 10mg and the dietary vitamin E levels were 0 and 150mg per kg diet, respectively. Alcohol-administrated groups received drinking water solution containing 10% of ethanol from the 3-weeks of experimental periods. The obtained experimental results are summarized as follow: The ${\gamma}$-GTP activity in plasma was higher in alcohol administrated groups and high selenium group (HSe) and low selenium group (LSe) than in control groups (CSe). The ${\gamma}$-GOT and GPT activities were higher in alcohol groups. The ${\gamma}$-GTP activity was significantly influenced by alcohol in LSe groups than in other groups. The glutathione peroxidase (GSH-Px) activity of plasma was significantly lower in LSe groups than HSe and CSe groups. The GSH-Px activity of microsomal and cytosolic fraction was slightly lower in alcohol groups and was about a half value lower in HSe and LSe groups than CSe groups. There was negative correlation between plasma Se level and GSH-Px activity of cytosolic fraction in HSe groups (r=- 0.662, p<0.001) and positive correlation in LSe groups (r=0.640, p<0.001). The GSH S-transferase activity in microsomal and cytosolic fraction was slightly higher in alcohol administrated but vitamin E nonadministrated groups, and significantly higher in LSe groups than in other groups. The catalase activity in mitochondria was lower in HSe than CSe groups, but rather higher in LSe groups. The superoxide dismutase (SOD) activity in cytosolic fraction of liver was not found any effect in all groups. The cytochrome P-450 was higher in alcohol groups, but significantly lower in HSe groups. In conclusion, the deficiency of Se and vitamin E develops the hyperoxidation of liver lipid through the increase of activity of enzyme related to the lipid peroxidation and alcohol administration appears to further increase of hyperoxidation of liver lipid.

  • PDF

Estimation of the Optimal Periods for Planting and Felling Larix kaempferi Based on the Period of Its Cambial Activity (형성층 활동 기간을 활용한 일본잎갈나무 식재 및 벌채 적기 추정)

  • YOO, Hye-Ji;JU, Jeong-Deuk;PARK, Jun-Hui;SHIN, Chang-Seob;EOM, Chang-Deuk;SEO, Jeong-Wook
    • Journal of the Korean Wood Science and Technology
    • /
    • v.49 no.5
    • /
    • pp.399-415
    • /
    • 2021
  • This study was conducted to suggest the optimal periods for planting and felling Larix kaempferi tree by monitoring its cambial activity period. In addition, the heat summation to induce the cambial activity of Larix kaempferi was investigated. The study sites were at Mt. Worak and Mt. Midong. After selecting 5 trees at Mt. Worak and Mt. Midong, the cambium samples were collected using a mini trephor with 2 mm diameter from April 7 to October 6, 2017 at Mt. Worak, and from April 7 to September 29, 2017 at Mt. Midong. After the collected cambium samples were embedded in PEG2000, transverse thin sections with a thickness of 10 to 15 ㎛ were prepared using a microtome. One Larix kaempferi sample from each site, Mt. Worak and Mt. Midong, in which the accurate monitoring of cambial activity was impossible due to the formation of traumatic resin canal, was excluded from the study. The observation of the initiation date of cambial activity under a light microscopy revealed that 2 specimens from Mt. Worak and 3 from Mt. Midong showed the initiation on April 28. The remaining 2 specimens of Mt. Worak and 1 specimen of Mt. Midong were initiated on May 4, which was a week later than the others. The heat summation that induced the initiation of cambial activity was 196.4-271.8 at Mt. Worak and 204.7-277.3 at Mt. Midong, which was similar. The termination of cambial activity occurred between August 4 and 25 at Mt. Worak, and between August 4 and September 1 at Mt. Midong. Based on the above results, it was found that the optimal planting period for Larix kaempferi in Mt. Worak and Mt. Midong was before April, about a month before the cambium activity, and the felling period was from October when the cambial activity was completely terminated.