Biomedical subjects

Jun Shiota

Publications and source records attributed to Jun Shiota.

6 recordsLinked to original sources

PubMed2006

Regulation of neurotrophin-3 gene transcription by Sp3 and Sp4 in neurons.

Neurotrophin-3 (NT-3), a neurotrophin member, plays crucial roles in neuronal development, function and plasticity. Previous studies have demonstrated that NT-3 gene transcription is driven by alternative promoters A and B, located upstream of exons 1A (EIA) and 1B (EIB), respectively. However, the transcription factors and DNA elements that drive NT-3 gene transcription remain to be identified. Here, we analysed the promoter region of the NT-3 gene and found that an NT-3 transcript containing EIB is predominantly expressed in cortical neurons which preferentially utilize promoter B, and two tandemly repeated GC-boxes, located between -100 and -60 base pairs within promoter B, are required for the transcription. Electrophoretic mobility shift and chromatin immunoprecipitation assays revealed that both specificity protein (Sp)3 and Sp4 were able to bind to the Sp1 binding sequences within the GC boxes. Expression of dominant-negative Sp3 and Sp4 small interfering RNA in cortical neurons reduced the activity of the NT-3 gene promoter. Over-expression of Sp1 family members, especially Sp4, resulted in an increase of the NT-3 gene promoter. These findings indicate that the NT-3 gene is a target gene for Sp4 that is abundantly expressed in the brain.

Animals↗

PubMed2006

Developmental expression of the SRF co-activator MAL in brain: role in regulating dendritic morphology.

The dynamic changes in dendritic morphology displayed by developing and mature neurons have stimulated interest in deciphering the signaling pathways involved. Recent studies have identified megakaryocytic acute leukemia (MAL), a serum response factor (SRF) co-activator, as a key component of a signaling pathway linking changes in the actin cytoskeleton to SRF-mediated transcription. To help define the role of this pathway in regulating dendritic morphology, we have characterized the pattern of MAL expression in the developing and adult brain, and have examined its role in regulating dendritic morphology in cultured cortical neurons. In histological studies of mouse brain, we found prominent expression of MAL in neurons in adult hippocampus and cerebral cortex. MAL immunostaining revealed localization of this protein in neuronal cell bodies and apical dendrites. During development, an increase in MAL expression occurs during the second post-natal week. Expression of dominant negative MAL constructs or MAL siRNA in cortical neurons grown in primary culture reduces the number of dendritic processes and decreases the basal level of SRF-mediated transcription. Taken together, these findings indicate that the MAL-SRF signaling pathway plays a key role in regulating dendritic morphology.

Animals↗

PubMed2005

Nuclear translocation of the SRF co-activator MAL in cortical neurons: role of RhoA signalling.

Although it is well established that RhoA signaling pathways play key roles in regulating neuronal morphology, their involvement in other aspects of neuronal function has received little attention. Recent studies have elucidated a novel intracellular signaling pathway used by RhoA to elicit activation of serum response factor (SRF)-mediated transcription. In this pathway, activation of RhoA triggers nuclear translocation of the SRF co-activator, megakaryocytic acute leukemia (MAL). In assessing whether RhoA regulates transcription in neurons via this pathway, we have found that a constitutively active form of Tech (transcript-enriched in cortex and hippocampus), a RhoA guanine nucleotide exchange factor (GEF) that is expressed in forebrain neurons, stimulates SRF reporter activity in extracts of primary cortical cultures and induces nuclear translocation of MAL in cortical neurons. Both of these responses appear to be mediated by Tech's activation of RhoA as they are not mimicked by a mutant Tech construct lacking RhoA GEF activity and are blocked by C3 transferase, a selective inhibitor of RhoA. Furthermore, Tech-induced increases in SRF activity are suppressed by a dominant negative MAL construct. These findings demonstrate that RhoA signaling pathways are able to regulate transcription in neurons by triggering translocation of the SRF co-activator MAL.

Active Transport, Cell Nucleus↗

PubMed2002

Down-regulation of CD43 molecule expression on intraperitoneal neutrophils in CAPD patients with peritonitis.

To assess the release of proteases from neutrophils infiltrated into the peritoneal cavity in continuous ambulatory peritoneal dialysis (CAPD), we investigated the regulation of CD43, LAM-1 and Mac-1 expression on the neutrophil plasma membrane using FACS analysis in CAPD patients with peritonitis. Five CAPD patients with peritonitis and five CAPD patients without peritonitis were studied. CD43 expression was immunohistochemically determined in both groups of patients using flow cytometry, and comparisons were made between the two groups. Down-regulation of CD43 and LAM-1, and up-regulation of Mac-1 were demonstrated on neutrophils obtained from CAPD dialysate of peritonitis patients after 1-h dwell time. Further up-regulation of Mac-1 developed until a dwell time of 4 h. Immunoblot analysis for neutrophil lysate from dialysate showed the presence of the asialo form of CD43 molecules and their fragments, which may be produced by cleavage of the CD43 molecule at extracellular sites. The intraperitoneal neutrophils in dialysate from CAPD patients with peritonitis are continuously activated during dwell time, and proteases may be released from neutrophils into dialysate after only a short dwell time.

Adult↗

PubMed2002

[A case with secondary hyperparathyroidism suggesting the direct suppressive effect of maxacalcitol on osteoblasts].

The direct effect of vitamin D on osteoblasts in secondary hyperparathyroidism(2 degrees HPT) is not necessarily obvious. We found an unusual hemodialyzed patient without any response to oral calcitriol pulse therapy(4 micrograms x 2/week), and who was administered maxacalcitol at a dose of up to 15 micrograms x 3/week for 28 months. Plasma intact parathyroid hormone(PTH) was not suppressed from the initial level of 1,773 pg/ml to 2,100 pg/ml. However, on the contrary, alkaline phosphatase(ALP) was successively suppressed from 2 weeks from the initial level of 1,261 IU/l to 276 IU/l. This result suggests a direct suppressive effect of maxacalcitol on osteoblasts in 2 degrees HPT.

Administration, Oral↗

PubMed2002

[Not Available].

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Journal Article↗