Smooth muscle protein 22α-Cre recombination in resting cardiac fibroblasts and hematopoietic precursors.

The Cre-loxP system has been widely used for cell- or organ-specific gene manipulation, but it is important to precisely understand what kind of cells the recombination takes place in. Smooth muscle 22α (SM22α)-Cre mice have been utilized to alter genes in vascular smooth muscle cells (VSMCs), activated fibroblasts or cardiomyocytes (CMs). Moreover, previous reports indicated that SM22α-Cre is expressed in adipocytes, platelets or myeloid cells. However, there have been no report of whether SM22α-Cre recombination takes place in nonCMs in hearts. Thus, we used the double-fluorescent Cre reporter mouse in which GFP is expressed when recombination occurs. Immunofluorescence analysis demonstrated that recombination occurred in resting cardiac fibroblasts (CFs) or macrophages, as well as VSMCs and CMs. Flow cytometry showed that some CFs, resident macrophages, neutrophils, T cells, and B cells were positive for GFP. These results prompted us to analyze bone marrow cells, and we observed GFP-positive hematopoietic precursor cells (HPCs). Taken together, these results indicated that SM22α-Cre-mediated recombination occurs in resting CFs and hematopoietic cell lineages, including HPCs, which is a cautionary point when using SM22α-Cre mice

Isoelectric Focusing Studies on Immunochemical Change with Age in Chicken Lens Crystallin

Changes of alpha-, beta- and delta-crystallins of chicken lens soluble protein with age from 7-day embryos to 1.5-year-old chickens were examined by isoelectric focusing and by immunochemical studies with anti chicken total lens crystallin serum and anti chicken beta-crystallin serum. Alpha- and delta-crystallins were isofocused in a narrow pI region (alpha : pi 4.90-5.30 ; delta : pI 4.90-6.60) and this range did not become broader with age. However, the pI range of beta-crystallin gradually became broader during development. On immunochemical analysis, beta-crystallin gave two types of precipitin line, named beta [Ls] and beta [Ls]. The beta [Ls] from 7-day embryos was split into two pI regions (pI 5.91-6.18 and pI 6.41-6.64), whereas that from 10-day embryos was fused into one line (pI 5.99-6.86) and a new precipitin line, beta [Lc], was detected at pI 6.65-6.86. These two types of precipitin line, beta [Ls]and beta [Lc], extended to an alkaline region before hatching, and to all pI regions (pI 4.26-8.19) in chickens of 40-60 days old. However, materials of over pI 7 of both beta [Ls] and beta [Lc] had disappeared in chickens of over 1.5 years old

Immunocytochemical Analysis of α-Tubulin Distribution Before and After Rapid Axopodial Contraction in the Centrohelid Raphidocystis contractilis

The centrohelid Raphidocystis contractilis is a heliozoan that has many radiating axopodia, each containing a bundle of microtubules. Although the rapid contraction of the axopodia at nearly a video rate (30 frames/s) is induced by mechanical stimuli, the mechanism underlying this phenomenon in R. contractilis has not yet been elucidated. In the present study, we described for the first time an adequate immunocytochemical fixation procedure for R. contractilis and the cellular distribution of α-tubulin before and after rapid axopodial contraction. We developed a flow-through chamber equipped with a micro-syringe pump that allowed the test solution to be injected at a flow rate below the threshold required to induce rapid axopodial contraction. Next, we used this injection method for evaluating the effects of different combinations of two fixatives (paraformaldehyde or glutaraldehyde) and two buffers (phosphate buffer or PHEM) on the morphological structure of the axopodia. A low concentration of glutaraldehyde in PHEM was identified as an adequate fixative for immunocytochemistry. The distribution of α-tubulin before and after rapid axopodial contraction was examined using immunocytochemistry and confocal laser scanning fluorescence microscopy. Positive signals were initially detected along the extended axopodia from the tips to the bases and were distributed in a non-uniform manner within the axopodia. Conversely, after the induction of a rapid axopodial contraction, these positive signals accumulated in the peripheral region of the cell. These results indicated that axopodial microtubules disassemble into fragments and/ or tubulin subunits during rapid axopodial contraction. Therefore, we hypothesize that the mechanism of extremely rapid axopodial contraction accompanied by cytoskeletal microtubule degradation in R. contractilis involves microtubule-severing at multiple sites

Cytoskeletal elements in an acoelomorph worm, Praesagittifera naikaiensis

Acoel flatworms can move in a variety of ways such as muscular and ciliary movements via cytoskeletal elements and their neural regulations. However, those locomotive mechanisms have not yet been fully elucidated. In this study, we examined the distribution of cytoskeletal elements including filamentous actin (F-actin) and tubulin, and the neuroanatomical organization in an acoelomorph worm, Praesagittifera naikaiensis (P. naikaiensis). Video microscopy revealed the elongation/contraction and the bending/rotation processes, and the ciliary gliding movement of P. naikaiensis. Histochemical and morphological analysis demonstrated that F-actin networks of inner longitudinal and outer circular muscle fibers were positioned along the entire surface of the body, and that the average distance between the circular muscle fibers in the contracted organism was decreased in the anterior region compared with that in the elongated organism. Electron microscopy showed dense bodies on the muscle cells of P. naikaiensis, which indicates that those muscle cells have the appearance of vertebrate smooth muscle cells. Immunohistochemical analysis revealed that -tubulin-positive signals on the ciliary microtubules had close contact with the F-actin network, and that neurite bundles labelled with anti dSap47 antibody as a neuronal marker run along the anterior-posterior body axis. These results indicate that the well-organized cytoskeletal elements and their neural control systems are preserved in P. naikaiensis, and that their mechanisms involved in those regulation systems are similar to those vertebrate systems. Further studies are needed to clarify the physiological mechanisms underlying the muscular and ciliary movements in P. naikaiensis

Structure and function of tegmentum vasculosum in avian cochlea

In spite of the importance of endocochlear DC potential (EP) and the K+-rich endolymph for the avian cochlea, the structure and function of the tegmentum vasculosum (TV) has not yet been fully elucidated, compared with those of the stria vascularis in the mammalian cochlea. In this study, we examined structural analysis of the epithelial cells, gene expressions of Na+-K+-ATPase (Atp1A1) and Kir4.1 (Kcnj10), and protein localizations of Na+-K+-ATPase and Kir4.1, in the TV. Tight junctional structures were observed between epithelial dark cells and light cells in the TV. Both Atp1a1 and Kcnj10 genes were detected in the TV. In addition, immunopositive signals for both Na+-K+-ATPase and Kir4.1 were recognized in the TV. These results indicate that Na+-K+-ATPase and Kir4.1 play roles in maintaining the EP and high K+ concentration of the endolymph. Further studies are needed to clarify the physiological functions of the TV. This is the first report which demonstrates that gene and protein expression data contribute to the avian inner ear homeostasis

Living alone is associated with depression among the elderly in a rural community in Japanp syg_402 179..185

Abstract Aim: This study aimed to investigate factors associated with depression in a sample of elderly Japanese individuals in a rural community and to examine differences among factors associated with individuals living alone or living with others. Methods: Using a population-based sample from rural Japan, we assessed a total of 1552 participants aged 65 years or older by mailing a survey and evaluating responses based on the Geriatric Depression Scale. Factors associated with depression were also examined. Results: We received 964 valid responses. Depressed subjects comprised 20.5% of the sample. Living alone was significantly related to depression. In individuals living alone, depression was associated with loss of appetite, suicidal ideation, financial strain, and worries in life. However, multiple linear regression analyses revealed that the influence of living alone was negated by having a good social support system. Conclusion: These findings confirm that living alone is an important factor in depression among the elderly in a rural part of Japan. Results also confirm what others have found in Western cultures: high levels of social support, awareness of receiving social support, and willingness to receive assistance may reduce the risk of depression

A safe combined nephrectomy and right lobectomy using the liver hanging maneuver for huge renal cell carcinoma directly invading the right lobe of the liver: report of a case

We herein discuss a patient who underwent simultaneous combined right nephrectomy and right lobectomy of the liver. A 64-year-old male was diagnosed with a huge right renal cell carcinoma (RCC), 13 cm in diameter, which was invading directly into the right hepatic lobe. This type of RCC has been rarely reported, and an anterior approach using the liver hanging maneuver was extremely useful during hepatic parenchymal dissection. The liver parenchymal dissection was performed prior to mobilization of the liver, because the mobilization of the right lobe of the liver was impossible. During the hepatic parenchymal resection, the liver was suspended with the tape and transected, and thereafter, retroperitoneal dissection, nephrectomy and right lobectomy of the liver were completed. The patient was discharged from the hospital on the 12th postoperative day with an uneventful clinical course. The anterior approach using the liver hanging maneuver during hepatic parenchymal resection can be safe and feasible for huge RCC invading the right hepatic lobe

Immunohistochemical study of the Landolt\u27s club cells in the chicken retina

The immunohistochemical specificity of the Landolt\u27s club cells of the chicken retina were studied by the indirect immunofluorescence and immunoperoxidase bridge methods with specific antiserum against neuronal filament proteins of these cells. In the tissue, the Landolt\u27s club cells were selectively stained with the specific antiserum (antigen : isoelectric point=6.29, molecular weight=69,000). It appeared that the cells were bipolar and that the cell body lay in the superficial part of the internal nuclear layer. However, it was difficult to observe the dendrites like those of typical bipolar cells which synapse with photoreceptors. No reaction was found in ganglion cells, amacrine cells, horizontal cells and other types of bipolar cells which occupy the zone just inside the external plexiform layer. From the 6th day of tissue culture, the selectively stained bipolar cells appeared on the surface of the monolayer cells. They were small and oval with large, spherical and eccentric nuclei. The cytoplasm was slightly less dense than other neuron cells