著者
Satoshi Yonezawa Momoko Shigematsu Kazuto Hirata Kensuke Hayashi
出版者
日本組織細胞化学会
雑誌
ACTA HISTOCHEMICA ET CYTOCHEMICA (ISSN:00445991)
巻号頁・発行日
vol.48, no.5, pp.145-152, 2015-10-29 (Released:2015-10-29)
参考文献数
25
被引用文献数
1 18

It has been recently reported that the centrosome of neurons does not have microtubule nucleating activity. Microtubule nucleation requires γ-tubulin as well as its recruiting proteins, GCP-WD/NEDD1 and CDK5RAP2 that anchor γ-tubulin to the centrosome. Change in the localization of these proteins during in vivo development of brain, however, has not been well examined. In this study we investigate the localization of γ-tubulin, GCP-WD and CDK5RAP2 in developing cerebral and cerebellar cortex with immunofluorescence. We found that γ-tubulin and its recruiting proteins were localized at centrosomes of immature neurons, while they were lost at centrosomes in mature neurons. This indicated that the loss of microtubule nucleating activity at the centrosome of neurons is due to the loss of γ-tubulin-recruiting proteins from the centrosome. RT-PCR analysis revealed that these proteins are still expressed after birth, suggesting that they have a role in microtubule generation in cell body and dendrites of mature neurons. Microtubule regrowth experiments on cultured mature neurons showed that microtubules are nucleated not at the centrosome but within dendrites. These data indicated the translocation of microtubule-organizing activity from the centrosome to dendrites during maturation of neurons, which would explain the mixed polarity of microtubules in dendrites.
著者
LANGE Ekkehard METZGER Martin BRAUN Katharina LUPPA Hans POEGGEL Gerd
出版者
日本組織細胞化学会
雑誌
Acta histochemica et cytochemica (ISSN:00445991)
巻号頁・発行日
vol.30, no.5, pp.505-512, 1997-10-01
参考文献数
34
被引用文献数
2 5

The cellular and spatial distribution of the presumptive retrograde transmitter nitric oxide, detected by NADPH-diaphorase histochemistry and of immunocytochemically labeled monoaminergic fiber systems was compared in regions of the medial prefrontal cortex of the precocious rodent Octodon degus. The staining patterns at two postnatal stages (PO and P14) were compared to those found in adult animals (P90). At birth, NADPH-diaphorase positive neurons were found in all cortical layers of the anterior cingulate, infralimbic and prelimbic cortex. During postnatal development the number of diaphorase-positive cells gradually decreased in layers II-VI and remained unchanged in layer I of these cortical regions. All NADPH-diaphorase positive neurons appeared to be spineless and pyramidal cells never contained NADPH-diaphorase. No colocalization of NADPH-diaphorase with either serotonin or TH was detectable. However, at all developmental stages the somata and proximal dendritic shafts of some of the NADPH-diaphorase containing bi-or multipolar neurons in layers V-VI were contacted by serotonin-and tyrosine hydroxylase-immunoreactive fibers. In a subpopulation of GABAergic interneurons in the deeper layers (V-VI) NADPH-diaphorase activity was colocalized with calbindin-D 28 k immunoreactivity.<BR>The abundance of nitridergic systems in regions of the medial prefrontal cortex already at birth together with their close spatial relationship with monoaminergic afferent systems as well as local GABAergic units may indicate a yet to determine role of these transmitter systems in early learning.
著者
Seiko Toyozawa Chikako Kaminaka Fukumi Furukawa Yasushi Nakamura Hiroshi Matsunaka Yuki Yamamoto
出版者
日本組織細胞化学会
雑誌
ACTA HISTOCHEMICA ET CYTOCHEMICA (ISSN:00445991)
巻号頁・発行日
vol.45, no.5, pp.293-299, 2012 (Released:2012-10-31)
参考文献数
22
被引用文献数
11 18

The CXCR4/CXCL12 pathway has recently been reported to be involved in stimulating the metastasis of many different neoplasms, in which CXCR4 activates various phenomena such as chemotaxis, invasion, angiogenesis and proliferation. The purpose of this study was to analyze a possible association between the expression of chemokine receptors CXCR4, CCR6 and CCR7 with the clinicopathological features of cutaneous malignant melanoma, and to assess the usefulness of these chemokine receptors for diagnosis and prognosis. In our study, a percentage of immunoexpression of both CXCR4 and its ligands CXCL12 was associated with high clinical risk. In contrast, the patients with a low immunoexpression of CXCR4 and CXCL12 had low clinical risk. CCR6 and CCR7 immunoexpressions were also correlated with some clinical parameters, but seemed no more useful than CXCR4. These data suggest that the assessment of CXCR4 immunoexpression is a novel tool for predicting tumor aggressiveness in malignant melanomas, and in particular, a high immunoexpression percentage of CXCR4 and CXCL12 might be a sign of a poor prognosis.
著者
貝谷 壽宣 難波 益之 加藤 一夫 森 秀樹
出版者
日本組織細胞化学会
雑誌
日本組織細胞化学会総会プログラムおよび抄録集
巻号頁・発行日
no.19, 1978-11-01

ヒト脳の黒質と青斑核の神経細胞には、生後4〜5年からメラニン色素(M)がみられ、加令と共に増加する。本研究ではこれらカテコールアミン産生ノイロンの活動状況を知る目的で、上記部位のM量を定量した。対象は12-82歳の明らかな神経精神疾患を認めなかった40剖検脳を用いた。ホルマリン固定後中脳および橋の中央部を水平断し、パラフィン包埋標本を作製し、各ブロック毎に10μm切片を2枚作り、1枚はニツスル染色を施し細胞がクループの判定に、他の1枚は無染色のまま封入した。無染色標本の黒質と青斑核ではMのみが茶褐色顆粒として検鏡される。この顆粒を走査型顕微濃度計(Nikon Vickers M85)にて定性すると550nm付近にて最大吸光度を示したので、この波長を使用し、バックグランドセット法でMを定量した。1回のスキャンニングに10〜25個の神経細胞が測定できるようシャドウイングした。スキャンニング時間5秒で3回測定し、それを和し、density値をarea値で除した値を各個体間の比較に使用した。対象脳では黒質でも青斑核においてもMは加令とともに増加した。(r=0.72、P<0.001)。
著者
Erina Ikeda Tetsuya Goto Kaori Gunjigake Kayoko Kuroishi Masae Ueda Shinji Kataoka Takashi Toyono Mitsushiro Nakatomi Yuji Seta Chiaki Kitamura Tatsuji Nishihara Tatsuo Kawamoto
出版者
日本組織細胞化学会
雑誌
ACTA HISTOCHEMICA ET CYTOCHEMICA (ISSN:00445991)
巻号頁・発行日
pp.15022, (Released:2016-02-10)
被引用文献数
5

Several theories have been proposed regarding pain transmission mechanisms in tooth. However, the exact signaling mechanism from odontoblasts to pulp nerves remains to be clarified. Recently, ATP-associated pain transmission has been reported, but it is unclear whether ATP is involved in tooth pain transmission. In the present study, we focused on the vesicular nucleotide transporter (VNUT), a transporter of ATP into vesicles, and examined whether VNUT was involved in ATP release from odontoblasts. We examined the expression of VNUT in rat pulp by RT-PCR and immunostaining. ATP release from cultured odontoblast-like cells with heat stimulation was evaluated using ATP luciferase methods. VNUT was expressed in pulp tissue, and the distribution of VNUT-immunopositive vesicles was confirmed in odontoblasts. In odontoblasts, some VNUT-immunopositive vesicles were colocalized with membrane fusion proteins. Additionally P2X3, an ATP receptor, immunopositive axons were distributed between odontoblasts. The ATP release by thermal stimulation from odontoblast-like cells was inhibited by the addition of siRNA for VNUT. These findings suggest that cytosolic ATP is transported by VNUT and that the ATP in the vesicles is then released from odontoblasts to ATP receptors on axons. ATP vesicle transport in odontoblasts seems to be a key mechanism for signal transduction from odontoblasts to axons in the pulp.
著者
Ryoko Yamagishi Tomohiko Wakayama Hiroki Nakata Kannika Adthapanyawanich Tewarat Kumchantuek Miyuki Yamamoto Shoichi Iseki
出版者
日本組織細胞化学会
雑誌
ACTA HISTOCHEMICA ET CYTOCHEMICA (ISSN:00445991)
巻号頁・発行日
vol.47, no.3, pp.95-102, 2014-06-30 (Released:2014-06-28)
参考文献数
28
被引用文献数
2 10

In the major salivary glands of mice, acinar cells in the parotid gland (PG) are known to be the main site for the production of the digestive enzyme α-amylase, whereas α-amylase production in the submandibular gland (SMG) and sublingual gland (SLG), as well as the cell types responsible for α-amylase production, has been less firmly established. To clarify this issue, we examined the expression and localization of both the mRNA and protein of α-amylase in the major salivary glands of male and female mice by quantitative and histochemical methods. α-amylase mRNA levels were higher in the order of PG, SMG, and SLG. No sexual difference was observed in α-amylase mRNA levels in the PG and SLG, whereas α-amylase mRNA levels in the female SMG were approximately 30% those in the male SMG. Using in situ hybridization and immunohistochemistry, signals for α-amylase mRNA and protein were found to be strongly positive in acinar cells of the PG, serous demilune cells of the SLG, and granular convoluted tubule (GCT) cells of the male SMG, weakly positive in seromucous acinar cells of the male and female SMG, and negative in mucous acinar cells of the SLG. These results clarified that α-amylase is produced mainly by GCT cells and partly by acinar cells in the SMG, whereas it is produced exclusively by serous demilune cells in the SLG of mice.
著者
Kudo Akihiko Hirano Hiroshi
出版者
日本組織細胞化学会
雑誌
Acta histochemica et cytochemica (ISSN:00445991)
巻号頁・発行日
vol.32, no.3, pp.229-233, 1999
被引用文献数
5 6 8

In histochemistry, image analysis has become quite common and is now regarded as a powerful tool that gives us semi-quantitative and precise interpretation on the obtained image data. However, image analysis itself is a time-consuming and laborious work, and so researches in the laboratory desire to make this task proceed as automatically as possible. In NIH Image and Adobe Photoshop^<【○!R】> software running on Power Macintosh computers, processes by automation are available in both programs, though they are quite different in each software. By use of both of these programs, the analysis processes become easier to set up. To automatically process the data, the suitable preprocessing of digital image data is required. That is, image processing programs are equipped with many processing programs, such as those for filtering, defining the threshold of images, or masking uninteresting areas. Additionally, a method for adequate labeling of the target substances and controlled conditions for the image digitization are always helpful.
著者
雨宮 次生 吉田 秀彦
出版者
日本組織細胞化学会
雑誌
日本組織細胞化学会総会プログラムおよび抄録集
巻号頁・発行日
no.19, 1978-11-01

研究目的 : phosphorylaseは, 生体においてはグリコーゲンの分解にのみ作用し試験管内ではグリコーゲンの合成と分解に作用するとされている。本研究では, Phosphorylaseが生体においても合成系酵素として作用する可能性を追求する。方法 : ひよこ視細胞副錐体paraboloidにおいては, in vitroならびにin vivoにおいて組織化学的方法を用いてphosphorylase系より合成された多糖体顆粒と, UDPG系を経て合成された顆粒との間には, 明瞭な形態学的差がある。基質液中のUDPGとG・I・Pの量を変え, pHを変えて反応させた時に生ずる多糖体顆粒の形態を比較し, どの経路から多糖体が合成されたかを推定する。この実験をin vitroおよびin vivoで行い, 結果を比較して生体におけるphosphorylaseの合成酵素たりうる可能性を検討する。結果 : UDPGとG・I・P等量pH5.7〜7.4の間では, 常にG・I・Pから合成されたと考えられる大きな多糖体顆粒が生じる。G・I・PをUDPGの1/4量にしてpH7.4にすると, UDPG系より合成されたと考えられる細顆粒が合成される。in vitro, in vivo共に同一結果をえた。結論 : Phosphorylaseは, 生体においても条件によりグリコーゲン合成に関与する。