ACTA HISTOCHEMICA ET CYTOCHEMICA

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Immunohistochemical and Morphofunctional Studies of Skeletal Muscle Tissues with Electric Nerve Stimulation by In Vivo Cryotechnique

Yuki Fukasawa, Nobuhiko Ohno, Yurika Saitoh, Takeshi Saigusa, Jun Arit ...

Article type: Regular Article
2015 Volume 48 Issue 2 Pages 27-36
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: April 16, 2015

DOIhttps://doi.org/10.1267/ahc.14068

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In this study, morphological and immunohistochemical alterations of skeletal muscle tissues during persistent contraction were examined by in vivo cryotechnique (IVCT). Contraction of gastrocnemius muscles was induced by sciatic nerve stimulation. The IVCT was performed immediately, 3 min or 10 min after the stimulation start. Prominent ripples of muscle fibers or wavy deformation of sarcolemma were detected immediately after the stimulation, but they gradually diminished to normal levels during the stimulation. The relative ratio of sarcomere and A band lengths was the highest in the control group, but it immediately decreased to the lowest level and then gradually recovered at 3 min or 10 min. Although histochemical intensity of PAS reaction was almost homogeneous in muscle tissues of the control group or immediately after the stimulation, it decreased at 3 min or 10 min. Serum albumin was immunolocalized as dot-like patterns within some muscle fibers at 3 min stimulation. These patterns became more prominent at 10 min, and the dots got larger and saccular in some sarcoplasmic regions. However, IgG1 and IgM were immunolocalized in blood vessels under nerve stimulation conditions. Therefore, IVCT was useful to capture the morphofunctional and metabolic changes of heterogeneous muscle fibers during the persistent contraction.

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Localization of Core Planar Cell Polarity Proteins, PRICKLEs, in Ameloblasts of Rat Incisors: Possible Regulation of Enamel Rod Decussation

Sumio Nishikawa, Tadafumi Kawamoto

Article type: Regular Article
2015 Volume 48 Issue 2 Pages 37-45
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: April 16, 2015

DOIhttps://doi.org/10.1267/ahc.14046

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To confirm the possible involvement of planar cell polarity proteins in odontogenesis, one group of core proteins, PRICKLE1, PRICKLE2, PRICKLE3, and PRICKLE4, was examined in enamel epithelial cells and ameloblasts by immunofluorescence microscopy. PRICKLE1 and PRICKLE2 showed similar localization in the proliferation and secretory zones of the incisor. Immunoreactive dots and short rods in ameloblasts and stratum intermedium cells were evident in the proliferation to differentiation zone, but in the secretion zone, cytoplasmic dots decreased and the distal terminal web was positive for PRICKLE1 and PRICKLE2. PRICKLE3 and PRICKLE4 showed cytoplasmic labeling in ameloblasts and other enamel epithelial cells. Double labeling of PRICKLE2 with VANGL1, which is another planar cell polarity protein, showed partial co-localization. To examine the transport route of PRICKLE proteins, PRICKLE1 localization was examined after injection of a microtubule-disrupting reagent, colchicine, and was compared with CX43, which is a membrane protein transported as vesicles via microtubules. The results confirmed the retention of immunoreactive dots for PRICKLE1 in the cytoplasm of secretory ameloblasts of colchicine-injected animals, but fewer dots were observed in control animals. These results suggest that PRICKLE1 and PRICKLE2 are transported as vesicles to the junctional area, and are involved in pattern formation of distal junctional complexes and terminal webs of ameloblasts, further implying a role in the formed enamel rod arrangement.

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Localization of Rod Bipolar Cells in the Mammalian Retina Using an Antibody Against the α₁c L-type Ca²⁺ Channel

Yu-Jin Huh, Jae-Sik Choi, Chang-Jin Jeon

Article type: Regular Article
2015 Volume 48 Issue 2 Pages 47-52
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: April 24, 2015

DOIhttps://doi.org/10.1267/ahc.14049

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Bipolar cells transmit stimuli via graded changes in membrane potential and neurotransmitter release is modulated by Ca²⁺ influx through L-type Ca²⁺ channels. The purpose of this study was to determine whether the α₁c subunit of L-type voltage-gated Ca²⁺ channel (α₁c L-type Ca²⁺ channel) colocalizes with protein kinase C alpha (PKC-α), which labels rod bipolar cells. Retinal whole mounts and vertical sections from mouse, hamster, rabbit, and dog were immunolabeled with antibodies against PKC-α and α₁c L-type Ca²⁺ channel, using fluorescein isothiocyanate (FITC) and Cy5 as visualizing agents. PKC-α-immunoreactive cells were morphologically identical to rod bipolar cells as previously reported. Their cell bodies were located within the inner nuclear layer, dendritic processes branched into the outer plexiform layer, and axons extended into the inner plexiform layer. Immunostaining showed that α₁c L-type Ca²⁺ channel colocalized with PKC-α in rod bipolar cells. The identical expression of PKC-α and α₁c L-type Ca²⁺ channel indicates that the α₁c L-type Ca²⁺ channel has a specific role in rod bipolar cells, and the antibody against the α₁c L-type Ca²⁺ channel may be a useful marker for studying the distribution of rod bipolar cells in mouse, hamster, rabbit, and dog retinas.

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Characteristics of the Localization of Connexin 43 in Satellite Cells during Skeletal Muscle Regeneration In Vivo

Minenori Ishido, Norikatsu Kasuga

Article type: Regular Article
2015 Volume 48 Issue 2 Pages 53-60
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: April 24, 2015

DOIhttps://doi.org/10.1267/ahc.14056

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For myogenesis, new myotubes are formed by the fusion of differentiated myoblasts. In the sequence of events for myotube formation, intercellular communication through gap junctions composed of connexin 43 (Cx43) plays critical roles in regulating the alignment and fusion of myoblasts in advances of myotube formation in vitro. On the other hand, the relationship between the expression patterns of Cx43 and the process of myotube formation in satellite cells during muscle regeneration in vivo remains poorly understood. The present study investigated the relationship between Cx43 and satellite cells in muscle regeneration in vivo. The expression of Cx43 was detected in skeletal muscles on day 1 post-muscle injury, but not in control muscles. Interestingly, the expression of Cx43 was not localized on the inside of the basement membrane of myofibers in the regenerating muscles. Moreover, although the clusters of differentiated satellite cells, which represent a more advanced stage of myotube formation, were observed on the inside of the basement membrane of myofibers in regenerating muscles, the expression of Cx43 was not localized in the clusters of these satellite cells. Therefore, in the present study, it was suggested that Cx43 may not directly contribute to muscle regeneration via satellite cells.

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SCGB3A2 Inhibits Acrolein-Induced Apoptosis through Decreased p53 Phosphorylation

Reiko Kurotani, Reika Shima, Yuki Miyano, Satoshi Sakahara, Yoshie Mat ...

Article type: Regular Article
2015 Volume 48 Issue 2 Pages 61-68
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: April 24, 2015

DOIhttps://doi.org/10.1267/ahc.14065

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Chronic obstructive pulmonary disease (COPD), a major global health problem with increasing morbidity and mortality rates, is anticipated to become the third leading cause of death worldwide by 2020. COPD arises from exposure to cigarette smoke. Acrolein, which is contained in cigarette smoke, is the most important risk factor for COPD. It causes lung injury through altering apoptosis and causes inflammation by augmenting p53 phosphorylation and producing reactive oxygen species (ROS). Secretoglobin (SCGB) 3A2, a secretory protein predominantly present in the epithelial cells of the lungs and trachea, is a cytokine-like small molecule having anti-inflammatory, antifibrotic, and growth factor activities. In this study, the effect of SCGB3A2 on acrolein-related apoptosis was investigated using the mouse fibroblast cell line MLg as the first step in determining the possible therapeutic value of SCGB3A2 in COPD. Acrolein increased the production of ROS and phosphorylation of p53 and induced apoptosis in MLg cells. While the extent of ROS production induced by acrolein was not affected by SCGB3A2, p53 phosphorylation was significantly decreased by SCGB3A2. These results demonstrate that SCGB3A2 inhibited acrolein-induced apoptosis through decreased p53 phosphorylation, not altered ROS levels.

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NOTE

Histochemical Analysis of Laminin α Chains in Diethylstilbestrol-Induced Prolactinoma in Rats

Dini Ramadhani, Alimuddin Tofrizal, Takehiro Tsukada, Takashi Yashiro

Article type: Note
2015 Volume 48 Issue 2 Pages 69-73
Published: April 28, 2015
Released on J-STAGE: April 28, 2015
Advance online publication: March 31, 2015

DOIhttps://doi.org/10.1267/ahc.14067

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Laminin, a major basement membrane component, is important in structural support and cell proliferation and differentiation. Its 19 isoforms are assemblies of α, β, and γ chains, and the α chains (α1-5) determine the isoform characteristics. Although our previous studies showed alterations in α chain expressions during anterior pituitary development, their expressions in ‍pituitary tumors yet to be determined. The present study used a rat model of diethylstilbestrol (DES)-induced prolactinoma to examine α chain expressions during prolactinoma tumorigenesis (0–12 weeks of DES treatment) by in situ hybridization and immunohistochemistry. mRNA of α1, α3, and α4 chains was detected in control and after 4 weeks of DES treatment. These expressions were undetectable after 8 weeks of DES treatment and in prolactinoma (12 weeks of DES treatment). Immunohistochemistry showed that the α1 chain was localized in some anterior pituitary cells in control and after 4 weeks of treatment and in endothelial cells after 8 weeks of treatment. The α3 and α4 chains were expressed in endothelial cells, and immunoreactivity and the number of immunopositive cells decreased during DES treatment. These findings suggest that alteration of laminin α chains is related to abnormal cell proliferation and neovascularization during development of DES-induced prolactinoma.

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