The Journal of Clinical Endocrinology & Metabolism, Oct 1, 2004
Both GH and IGF-I stimulate bone growth, but the molecular mechanisms mediating their effects on ... more Both GH and IGF-I stimulate bone growth, but the molecular mechanisms mediating their effects on the growth plate are not fully understood. We measured gene expression by microarray analysis in primary cultured human chondrocytes treated with either GH or IGF-I. One of the genes found to be up-regulated by both GH and IGF-I was that encoding cartilage oligomeric matrix protein (COMP). This protein is predominantly found in the extracellular matrix of cartilage. Mutations in the COMP gene have been associated with syndromes of short stature. To verify that COMP is regulated by GH in vivo, we measured COMP levels in serum in short children treated with GH. The study included 113 short pre-pubertal children (14 girls and 99 boys) with a mean (؎ SD) age of 8.84 ؎ 2.76 yr, height SD score of ؊2.74 ؎ 0.67, and IGF-I SD score of ؊1.21 ؎ 1.07 at the start of GH administration. Serum levels of COMP were 1.58 ؎ 0.28, 1.83 ؎ 0.28 (P < 0.0001), 1.91 ؎ 0.28 (P < 0.0001), 1.78 ؎ 0.28 (P < 0.001), and 1.70 ؎ 0.24 (P < 0.05) g/ml at baseline and after 1 wk and 1, 3, and 12 months, respectively. In conclusion, we have demonstrated that COMP expression is up-regulated by both GH and IGF-I in primary cultured human chondrocytes. Furthermore, serum levels of COMP increase after the start of GH treatment in short children.
Autophagy is a membrane traffic system that provides sustainable degradation of cellular componen... more Autophagy is a membrane traffic system that provides sustainable degradation of cellular components for homeostasis, and is thus considered to promote health and longevity, though its activity declines with aging. The present findings show deterioration of autophagy in association with premature skin aging. Autophagy flux was successfully determined in skin tissues, which demonstrated significantly decreased autophagy in hyperpigmented skin such as that seen in senile lentigo. Furthermore, an exacerbated decline in autophagy was confirmed in xerotic hyperpigmentation areas, accompanied by severe dehydration and a barrier defect, which showed correlations with skin physiological conditions. The enhancement of autophagy in skin ex vivo ameliorated skin integrity, including pigmentation and epidermal differentiation. The present results indicate that the restoration of autophagy can contribute to improving premature skin aging by various intrinsic and extrinsic factors via the normaliz...
The immune-regulatory compound histamine is involved in the metabolism of the essential skin comp... more The immune-regulatory compound histamine is involved in the metabolism of the essential skin component hyaluronan (HA). We previously reported that histamine up-regulates the expression of hyaluronan-binding protein involved in hyaluronan depolymerization (HYBID, aka CEMIP or KIAA1199), which plays a key role in HA degradation. However, no information is available about histamine’s effects on HA synthase (HAS) expression, the molecular sizes of the HAs produced, and histamine receptors and their signaling pathways in skin fibroblasts. Moreover, histamine’s effects on photoaged skin remain elusive. Here, we show that histamine increases HA degradation by up-regulating HYBID and down-regulating HAS2 in human skin fibroblasts in a dose- and time-dependent manner and thereby decreases the total amounts and sizes of newly produced HA. Histamine H1 blocker abrogated the histamine effects on HYBID up-regulation, HAS2 suppression, and HA degradation. Histamine H1 agonist exhibited effects o...
Rhododendrol is a phenolic compound that shows a tyrosinase-dependent toxicity for melanocytes an... more Rhododendrol is a phenolic compound that shows a tyrosinase-dependent toxicity for melanocytes and occasionally induces a vitiligo-like skin depigmentation. The post-tyrosinase mechanisms determining melanocyte death or survival, however, are far from clear. Here, we find that rhododendrol treatment leads to a reduction in the levels of cellular glutathione but also induces a cellular antioxidant response that eventually increases glutathione levels. We further find that rhododendrol toxicity is enhanced when glutathione levels are experimentally reduced and alleviated when glutathione levels are increased. Hence, it appears that the size of the preexisting glutathione pool along with the capacity to supply glutathione via the antioxidant response determines whether melanocytes survive or die after rhododendrol exposure. It is conceivable, therefore, that rhododendrol-induced leukoderma depends on the capacity to maintain appropriate glutathione levels and that enhancement of glutat...
Regulation of hyaluronan (HA) synthesis and degradation is essential to maintenance of extracellu... more Regulation of hyaluronan (HA) synthesis and degradation is essential to maintenance of extracellular matrix homeostasis. We recently reported that HYBID (HYaluronan-Binding protein Involved in hyaluronan Depolymerization), also called KIAA1199, plays a key role in HA depolymerization in skin and arthritic synovial fibroblasts. However, regulation of HA metabolism mediated by HYBID and HA synthases (HASs) under stimulation with growth factors remains obscure. Here we report that TGF-1, basic FGF, EGF, and PDGF-BB commonly enhance total amount of HA in skin fibroblasts through up-regulation of HAS expression, but molecular size of newly produced HA is dependent on HYBID expression levels. Stimulation of HAS1/2 expression and suppression of HYBID expression by TGF-1 were abrogated by blockade of the MAPK and/or Smad signaling and the PI3K-Akt signaling, respectively. In normal human skin, expression of the TGF-1 receptors correlated positively with HAS2 expression and inversely with HYBID expression. On the other hand, TGF-1 upregulated HAS1/2 expression but exerted only a slight suppressive effect on HYBID expression in synovial fibroblasts from the patients with osteoarthritis or rheumatoid arthritis, resulting in the production of lower molecular weight HA compared with normal skin and synovial fibroblasts. These data demonstrate that although TGF-1, basic FGF, EGF, and PDGF-BB enhance HA production in skin fibroblasts, TGF-1 most efficiently contributes to production of high molecular weight HA by HAS up-regulation and HYBID down-regulation and suggests that inefficient downregulation of HYBID by TGF-1 in arthritic synovial fibroblasts may be linked to accumulation of depolymerized HA in synovial fluids in arthritis patients.
Rhododendrol, an inhibitor of melanin synthesis developed for lightening/whitening cosmetics, was... more Rhododendrol, an inhibitor of melanin synthesis developed for lightening/whitening cosmetics, was recently reported to induce a depigmentary disorder principally at the sites of repeated chemical contact. Rhododendrol competitively inhibited mushroom tyrosinase and served as a good substrate, while it also showed cytotoxicity against cultured human melanocytes at high concentrations sufficient for inhibiting tyrosinase. The cytotoxicity was abolished by phenylthiourea, a chelator of the copper ions at the active site, and by specific knockdown of tyrosinase with siRNA. Hence, the cytotoxicity appeared to be triggered by the enzymatic conversion of rhododendrol to active product(s). No reactive oxygen species were detected in the treated melanocytes, but up-regulation of the CCAAT-enhancer-binding protein homologous protein gene responsible for apoptosis and/or autophagy and caspase-3 activation were found to be tyrosinase dependent. These results suggest that a tyrosinase-dependent ...
The granule cells (GCs) of dentate gyrus exhibit regionally specific morphology, and continue to ... more The granule cells (GCs) of dentate gyrus exhibit regionally specific morphology, and continue to be born and to develop well into adult life. We used a novel monoclonal antibody, MAb2G7, elicited by immunization of a mouse with a microsome fraction of the hippocampus, to evaluate regional and age-related differences in GCs immunohistochemically. Weak cytoplasmic reactions were observed in many neurons, but intense MAb2G7-positive dots were observed only in GCs. Using electron microscopy, we observed that these dots were localized in the internal droplets of secondary lysosome-like structures in GCs. The MAb2G7-positive granules were quantitatively analyzed in young adult and middle-aged rats. Larger numbers of reactive granules were observed in the infrapyramidal blade (IPB) than in the suprapyramidal blade (SPB) and the numbers of positive granules were proportionally reduced in the two areas in middle-aged rats. The changes in the MAb2G7 immunoreactivity may reflect different activation or neurogeneration of GCs in the IPB versus the SPB, and in middle-aged versus young adult rats.
The expression of cytoplasmic dynein and kinesin in rat Sertoli cells was examined using immunohi... more The expression of cytoplasmic dynein and kinesin in rat Sertoli cells was examined using immunohistochemistry with light and electron microscopies, and the existence of such bi-directional motors in the cells was confirmed. Cytoplasmic dynein was localized in the ectoplasmic specializations (ESs), mitochondria, and endoplasmic reticula. Among microtubules, an intense reaction of dynein was also detected, while, the reaction of kinesin changed in intensity in a stagedependent manner. The most distinct reaction of kinesin in Sertoli cells was detected at stages VII-VIII, while a weak reaction was observed at stages IX-XIV. No positive reaction was detected at stages I-VI. The intracellular localization of kinesin was detected in the ES (stage X) and endoplasmic reticulum, but a defined reaction was not observed among microtubules. These results suggested that an active bi-directional organelle transport is performed in rat Sertoli cells, and the system may relate to the elongation and release of spermatids, and to the migration of membranous organelles such as mitochondria and endoplasmic reticula. In addition, the sliding movement of microtubules may be carried out mainly by cytoplasmic dynein.
Polarization-sensitive optical coherence tomography (PS-OCT) permits non-invasive visualization o... more Polarization-sensitive optical coherence tomography (PS-OCT) permits non-invasive visualization of dermal birefringence, mainly due to collagenous structures. The purpose of this study is to use PS-OCT to assess intrinsic-age-related and photo-age-related differences in three-dimensional dermal birefringence. We measured dermal birefringence of the cheek skin and photo-protected interior upper arm skin from old and young volunteers. The algorithm that we used automatically produces the transversal dermal birefringence map from the polarization-sensitive OCT volume. This allowed quantitative comparison and visualization of the transverse distribution of the dermal birefringence. We found that dermal birefringence of the cheek skin was significantly smaller in the old group than in the young group (young group, 0.295±0.0371 mm À1 ; old group, 0.207±0.031 mm À1 ; P ¼ 0.003), whereas the interior upper arm showed no age-dependent difference. The transversal map of the cheek showed a heterogeneous decrease in dermal birefringence due to photoaging. The maps suggested that the peripheral regions of some infundibula were surrounded by a strong collagen network. Three-dimensional analyses of dermal birefringence using PS-OCT help to quantify the diagnosis of photoaging.
Background: Hyaluronan (HA) plays a role in keratinocyte proliferation and differentiation, and h... more Background: Hyaluronan (HA) plays a role in keratinocyte proliferation and differentiation, and have shown different biological activities depending on its molecular mass. While many studies have shown changes in the amount of HA after UVB irradiation, molecular mass change remains to be elucidated. Objective: To investigate the change in the molecular mass of HA after UVB irradiation in mouse epidermis. Methods: The mice were irradiated with a single dose of UVB (0.15 J/cm 2). The amount of HA was examined using HABP sandwich assay. The molecular mass distribution was estimated by Sephacryl S-1000 chromatography. Has and Hyal mRNA expressions were detected by real-time PCR. Results: On day 2 after UVB irradiation, both the amount of HA and the up-regulation of Has3 mRNA expression reached their maximum. The average HA molecular mass was about 1000 kDa, a level similar to that of the non-irradiated epidermis. On day 3, the average HA molecular mass drastically decreased to 100 kDa, while Hyal1, Hyal2, and Hyal3 mRNA expressions slightly increased. The amount of HA, however, remained high. On days 4 and 5, the amount of HA gradually decreased, but the molecular mass of HA remained low. A drastic reduction of the HA molecular mass after UVB irradiation was confirmed. Conclusion: UVB irradiation elicits remarkable changes in the molecular mass of HA, as well as amount. These qualitative and quantitative changes of HA might play an important role in UVB-induced cell proliferation and differentiation. Further study will be required to resolve the mechanism of HA degradation in the epidermis.
To clarify the anatomical location of medullary neurons associated with vomiting, the musk shrew ... more To clarify the anatomical location of medullary neurons associated with vomiting, the musk shrew (Suncus murinus), a small animal used as a model for emesis, was exposed to various emetic stimuli and patterns of neuronal excitation were investigated by Fos immunohistochemistry. In motion experiments, musk shrews were shaken for 30 min on a tabletop shaker (displacement = 25 mm and frequency = 1.2 Hz). Ten of fifteen animals vomited frequently (Mo-FV group); the other five animals did not vomit (Mo-NV group). In radiation experiments, X-ray irradiation (10 Gy) of the whole body caused frequent vomiting in all of seven experimental animals (Ra-FV group). In the Mo-FV group, many Fos-immunoreactive (Fos-ir) neurons were detected in the nucleus of the solitary tract (NTS) and the reticular formation. The distribution pattern of Fos-ir neurons in the Mo-NV group was similar to that in the Mo-FV group, but the Mo-NV group had significantly fewer positive neurons in the NTS and the reticular formation around the nucleus ambiguus. In the Ra-FV group, numerous Fos-ir neurons were observed in the area postrema, an area containing no positive neurons in the motion-stimulated animals. The number of Fos-ir neurons in the NTS of the Ra-FV group was not statistically different from that of the Mo-NV group. In the Mo-FV and Ra-FV groups, Fos-ir neurons were clustered in the reticular formation at the dorsal-dorsomedial edge of the nucleus ambiguus at the level of the rostral medulla, while few such clusters were observed in the Mo-NV group. These neurons may play a role in the regulation of the vomiting response.
Cardiac vagal preganglionic neurons (CVN) control cardiac activity by negative chronotropic, drom... more Cardiac vagal preganglionic neurons (CVN) control cardiac activity by negative chronotropic, dromotropic and inotropic effects. We attempted to characterize the distribution and neuronal properties of the CVN by using double labeling with the retrograde tracer cholera toxin B subunit (CTb) and immunohistochemistry for choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP) or nitric oxide synthase (NOS). Injection of CTb into the sinoatrial ganglia resulted in many retrogradely labeled of neurons in the dorsal motor nucleus of the vagus (DMV), the compact (AmC), semicompact (AmS), loose (AmL), external (AmE) formations of the nucleus ambiguus, and the intermediate zone (IZ) between DMV and the nucleus ambiguus. Almost all CTb-labeled neurons showed ChAT immunoreactivity in the DMV, AmC, AmS, AmL and IZ, but most of the CTb-labeled neurons showed no ChAT immunoreactivity in the AmE. Most of the CTb-labeled neurons were double-labeled with CGRP immunoreactivity in the AmC, AmS and AmL, but a few doublelabeled neurons were found in the DMV, IZ and AmE. A few CTb-labeled neurons were double-labeled with NOS immunoreactivity only in the DMV. No TH-immunoreactive neurons were found among the CVN. These results indicate that there are four kinds of neurons among the CVN: non-cholinergic CVN in the AmE, cholinergic and CGRP-containing CVN in the AmC, AmS and AmL, and cholinergic or cholinergic and NOS-containing CVN in the DMV.
The visual cortex in the rat is matured physiologically by postnatal day 30, but the visual syste... more The visual cortex in the rat is matured physiologically by postnatal day 30, but the visual system retains the potential to be reorganized until postnatal day 45. Therefore, we defined the period from postnatal days 28-45 as the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;late critical phase&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;. To examine whether monocular deprivation during the late critical phase gives rise to neuronal apoptosis in the dorsal lateral geniculate nucleus (dLGN), we used the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling method and anterograde tracing. The number of apoptotic cells in the dLGN after monocular deprivation at postnatal day 28 showed little difference from control at postnatal day 29, but was significantly increased to more than fourfold of control ipsilaterally to the monocular deprivation at postnatal day 35 and to more than 10-fold of control bilaterally at postnatal day 40. In the control, there were almost no apoptotic neurons in the dLGN on either side at postnatal day 40. In the nucleus ipsilateral to the monocular deprivation, approximately half the apoptotic neurons were found in an area that did not receive a retinal projection. These findings suggest that the biological process of increased apoptosis in the dLGN of rats that received monocular deprivation in the late critical phase may be different from that in the early critical phase. The increased number of apoptotic cells in the dLGN in the late critical phase may not be simply the result of monocular deprivation.
Biochimica Et Biophysica Acta-proteins and Proteomics, 2011
It is well known that advanced glycation end products (AGEs) are formed in long-lived dermal prot... more It is well known that advanced glycation end products (AGEs) are formed in long-lived dermal proteins such as collagen, and that their formation is related to skin aging. To examine the distribution of AGEs in skin tissue, we performed immunofluorescence studies on the human skin using an anti-AGEs antibody. Interestingly, AGEs signals were observed not only in the dermis but
The Journal of Clinical Endocrinology & Metabolism, Oct 1, 2004
Both GH and IGF-I stimulate bone growth, but the molecular mechanisms mediating their effects on ... more Both GH and IGF-I stimulate bone growth, but the molecular mechanisms mediating their effects on the growth plate are not fully understood. We measured gene expression by microarray analysis in primary cultured human chondrocytes treated with either GH or IGF-I. One of the genes found to be up-regulated by both GH and IGF-I was that encoding cartilage oligomeric matrix protein (COMP). This protein is predominantly found in the extracellular matrix of cartilage. Mutations in the COMP gene have been associated with syndromes of short stature. To verify that COMP is regulated by GH in vivo, we measured COMP levels in serum in short children treated with GH. The study included 113 short pre-pubertal children (14 girls and 99 boys) with a mean (؎ SD) age of 8.84 ؎ 2.76 yr, height SD score of ؊2.74 ؎ 0.67, and IGF-I SD score of ؊1.21 ؎ 1.07 at the start of GH administration. Serum levels of COMP were 1.58 ؎ 0.28, 1.83 ؎ 0.28 (P < 0.0001), 1.91 ؎ 0.28 (P < 0.0001), 1.78 ؎ 0.28 (P < 0.001), and 1.70 ؎ 0.24 (P < 0.05) g/ml at baseline and after 1 wk and 1, 3, and 12 months, respectively. In conclusion, we have demonstrated that COMP expression is up-regulated by both GH and IGF-I in primary cultured human chondrocytes. Furthermore, serum levels of COMP increase after the start of GH treatment in short children.
Autophagy is a membrane traffic system that provides sustainable degradation of cellular componen... more Autophagy is a membrane traffic system that provides sustainable degradation of cellular components for homeostasis, and is thus considered to promote health and longevity, though its activity declines with aging. The present findings show deterioration of autophagy in association with premature skin aging. Autophagy flux was successfully determined in skin tissues, which demonstrated significantly decreased autophagy in hyperpigmented skin such as that seen in senile lentigo. Furthermore, an exacerbated decline in autophagy was confirmed in xerotic hyperpigmentation areas, accompanied by severe dehydration and a barrier defect, which showed correlations with skin physiological conditions. The enhancement of autophagy in skin ex vivo ameliorated skin integrity, including pigmentation and epidermal differentiation. The present results indicate that the restoration of autophagy can contribute to improving premature skin aging by various intrinsic and extrinsic factors via the normaliz...
The immune-regulatory compound histamine is involved in the metabolism of the essential skin comp... more The immune-regulatory compound histamine is involved in the metabolism of the essential skin component hyaluronan (HA). We previously reported that histamine up-regulates the expression of hyaluronan-binding protein involved in hyaluronan depolymerization (HYBID, aka CEMIP or KIAA1199), which plays a key role in HA degradation. However, no information is available about histamine’s effects on HA synthase (HAS) expression, the molecular sizes of the HAs produced, and histamine receptors and their signaling pathways in skin fibroblasts. Moreover, histamine’s effects on photoaged skin remain elusive. Here, we show that histamine increases HA degradation by up-regulating HYBID and down-regulating HAS2 in human skin fibroblasts in a dose- and time-dependent manner and thereby decreases the total amounts and sizes of newly produced HA. Histamine H1 blocker abrogated the histamine effects on HYBID up-regulation, HAS2 suppression, and HA degradation. Histamine H1 agonist exhibited effects o...
Rhododendrol is a phenolic compound that shows a tyrosinase-dependent toxicity for melanocytes an... more Rhododendrol is a phenolic compound that shows a tyrosinase-dependent toxicity for melanocytes and occasionally induces a vitiligo-like skin depigmentation. The post-tyrosinase mechanisms determining melanocyte death or survival, however, are far from clear. Here, we find that rhododendrol treatment leads to a reduction in the levels of cellular glutathione but also induces a cellular antioxidant response that eventually increases glutathione levels. We further find that rhododendrol toxicity is enhanced when glutathione levels are experimentally reduced and alleviated when glutathione levels are increased. Hence, it appears that the size of the preexisting glutathione pool along with the capacity to supply glutathione via the antioxidant response determines whether melanocytes survive or die after rhododendrol exposure. It is conceivable, therefore, that rhododendrol-induced leukoderma depends on the capacity to maintain appropriate glutathione levels and that enhancement of glutat...
Regulation of hyaluronan (HA) synthesis and degradation is essential to maintenance of extracellu... more Regulation of hyaluronan (HA) synthesis and degradation is essential to maintenance of extracellular matrix homeostasis. We recently reported that HYBID (HYaluronan-Binding protein Involved in hyaluronan Depolymerization), also called KIAA1199, plays a key role in HA depolymerization in skin and arthritic synovial fibroblasts. However, regulation of HA metabolism mediated by HYBID and HA synthases (HASs) under stimulation with growth factors remains obscure. Here we report that TGF-1, basic FGF, EGF, and PDGF-BB commonly enhance total amount of HA in skin fibroblasts through up-regulation of HAS expression, but molecular size of newly produced HA is dependent on HYBID expression levels. Stimulation of HAS1/2 expression and suppression of HYBID expression by TGF-1 were abrogated by blockade of the MAPK and/or Smad signaling and the PI3K-Akt signaling, respectively. In normal human skin, expression of the TGF-1 receptors correlated positively with HAS2 expression and inversely with HYBID expression. On the other hand, TGF-1 upregulated HAS1/2 expression but exerted only a slight suppressive effect on HYBID expression in synovial fibroblasts from the patients with osteoarthritis or rheumatoid arthritis, resulting in the production of lower molecular weight HA compared with normal skin and synovial fibroblasts. These data demonstrate that although TGF-1, basic FGF, EGF, and PDGF-BB enhance HA production in skin fibroblasts, TGF-1 most efficiently contributes to production of high molecular weight HA by HAS up-regulation and HYBID down-regulation and suggests that inefficient downregulation of HYBID by TGF-1 in arthritic synovial fibroblasts may be linked to accumulation of depolymerized HA in synovial fluids in arthritis patients.
Rhododendrol, an inhibitor of melanin synthesis developed for lightening/whitening cosmetics, was... more Rhododendrol, an inhibitor of melanin synthesis developed for lightening/whitening cosmetics, was recently reported to induce a depigmentary disorder principally at the sites of repeated chemical contact. Rhododendrol competitively inhibited mushroom tyrosinase and served as a good substrate, while it also showed cytotoxicity against cultured human melanocytes at high concentrations sufficient for inhibiting tyrosinase. The cytotoxicity was abolished by phenylthiourea, a chelator of the copper ions at the active site, and by specific knockdown of tyrosinase with siRNA. Hence, the cytotoxicity appeared to be triggered by the enzymatic conversion of rhododendrol to active product(s). No reactive oxygen species were detected in the treated melanocytes, but up-regulation of the CCAAT-enhancer-binding protein homologous protein gene responsible for apoptosis and/or autophagy and caspase-3 activation were found to be tyrosinase dependent. These results suggest that a tyrosinase-dependent ...
The granule cells (GCs) of dentate gyrus exhibit regionally specific morphology, and continue to ... more The granule cells (GCs) of dentate gyrus exhibit regionally specific morphology, and continue to be born and to develop well into adult life. We used a novel monoclonal antibody, MAb2G7, elicited by immunization of a mouse with a microsome fraction of the hippocampus, to evaluate regional and age-related differences in GCs immunohistochemically. Weak cytoplasmic reactions were observed in many neurons, but intense MAb2G7-positive dots were observed only in GCs. Using electron microscopy, we observed that these dots were localized in the internal droplets of secondary lysosome-like structures in GCs. The MAb2G7-positive granules were quantitatively analyzed in young adult and middle-aged rats. Larger numbers of reactive granules were observed in the infrapyramidal blade (IPB) than in the suprapyramidal blade (SPB) and the numbers of positive granules were proportionally reduced in the two areas in middle-aged rats. The changes in the MAb2G7 immunoreactivity may reflect different activation or neurogeneration of GCs in the IPB versus the SPB, and in middle-aged versus young adult rats.
The expression of cytoplasmic dynein and kinesin in rat Sertoli cells was examined using immunohi... more The expression of cytoplasmic dynein and kinesin in rat Sertoli cells was examined using immunohistochemistry with light and electron microscopies, and the existence of such bi-directional motors in the cells was confirmed. Cytoplasmic dynein was localized in the ectoplasmic specializations (ESs), mitochondria, and endoplasmic reticula. Among microtubules, an intense reaction of dynein was also detected, while, the reaction of kinesin changed in intensity in a stagedependent manner. The most distinct reaction of kinesin in Sertoli cells was detected at stages VII-VIII, while a weak reaction was observed at stages IX-XIV. No positive reaction was detected at stages I-VI. The intracellular localization of kinesin was detected in the ES (stage X) and endoplasmic reticulum, but a defined reaction was not observed among microtubules. These results suggested that an active bi-directional organelle transport is performed in rat Sertoli cells, and the system may relate to the elongation and release of spermatids, and to the migration of membranous organelles such as mitochondria and endoplasmic reticula. In addition, the sliding movement of microtubules may be carried out mainly by cytoplasmic dynein.
Polarization-sensitive optical coherence tomography (PS-OCT) permits non-invasive visualization o... more Polarization-sensitive optical coherence tomography (PS-OCT) permits non-invasive visualization of dermal birefringence, mainly due to collagenous structures. The purpose of this study is to use PS-OCT to assess intrinsic-age-related and photo-age-related differences in three-dimensional dermal birefringence. We measured dermal birefringence of the cheek skin and photo-protected interior upper arm skin from old and young volunteers. The algorithm that we used automatically produces the transversal dermal birefringence map from the polarization-sensitive OCT volume. This allowed quantitative comparison and visualization of the transverse distribution of the dermal birefringence. We found that dermal birefringence of the cheek skin was significantly smaller in the old group than in the young group (young group, 0.295±0.0371 mm À1 ; old group, 0.207±0.031 mm À1 ; P ¼ 0.003), whereas the interior upper arm showed no age-dependent difference. The transversal map of the cheek showed a heterogeneous decrease in dermal birefringence due to photoaging. The maps suggested that the peripheral regions of some infundibula were surrounded by a strong collagen network. Three-dimensional analyses of dermal birefringence using PS-OCT help to quantify the diagnosis of photoaging.
Background: Hyaluronan (HA) plays a role in keratinocyte proliferation and differentiation, and h... more Background: Hyaluronan (HA) plays a role in keratinocyte proliferation and differentiation, and have shown different biological activities depending on its molecular mass. While many studies have shown changes in the amount of HA after UVB irradiation, molecular mass change remains to be elucidated. Objective: To investigate the change in the molecular mass of HA after UVB irradiation in mouse epidermis. Methods: The mice were irradiated with a single dose of UVB (0.15 J/cm 2). The amount of HA was examined using HABP sandwich assay. The molecular mass distribution was estimated by Sephacryl S-1000 chromatography. Has and Hyal mRNA expressions were detected by real-time PCR. Results: On day 2 after UVB irradiation, both the amount of HA and the up-regulation of Has3 mRNA expression reached their maximum. The average HA molecular mass was about 1000 kDa, a level similar to that of the non-irradiated epidermis. On day 3, the average HA molecular mass drastically decreased to 100 kDa, while Hyal1, Hyal2, and Hyal3 mRNA expressions slightly increased. The amount of HA, however, remained high. On days 4 and 5, the amount of HA gradually decreased, but the molecular mass of HA remained low. A drastic reduction of the HA molecular mass after UVB irradiation was confirmed. Conclusion: UVB irradiation elicits remarkable changes in the molecular mass of HA, as well as amount. These qualitative and quantitative changes of HA might play an important role in UVB-induced cell proliferation and differentiation. Further study will be required to resolve the mechanism of HA degradation in the epidermis.
To clarify the anatomical location of medullary neurons associated with vomiting, the musk shrew ... more To clarify the anatomical location of medullary neurons associated with vomiting, the musk shrew (Suncus murinus), a small animal used as a model for emesis, was exposed to various emetic stimuli and patterns of neuronal excitation were investigated by Fos immunohistochemistry. In motion experiments, musk shrews were shaken for 30 min on a tabletop shaker (displacement = 25 mm and frequency = 1.2 Hz). Ten of fifteen animals vomited frequently (Mo-FV group); the other five animals did not vomit (Mo-NV group). In radiation experiments, X-ray irradiation (10 Gy) of the whole body caused frequent vomiting in all of seven experimental animals (Ra-FV group). In the Mo-FV group, many Fos-immunoreactive (Fos-ir) neurons were detected in the nucleus of the solitary tract (NTS) and the reticular formation. The distribution pattern of Fos-ir neurons in the Mo-NV group was similar to that in the Mo-FV group, but the Mo-NV group had significantly fewer positive neurons in the NTS and the reticular formation around the nucleus ambiguus. In the Ra-FV group, numerous Fos-ir neurons were observed in the area postrema, an area containing no positive neurons in the motion-stimulated animals. The number of Fos-ir neurons in the NTS of the Ra-FV group was not statistically different from that of the Mo-NV group. In the Mo-FV and Ra-FV groups, Fos-ir neurons were clustered in the reticular formation at the dorsal-dorsomedial edge of the nucleus ambiguus at the level of the rostral medulla, while few such clusters were observed in the Mo-NV group. These neurons may play a role in the regulation of the vomiting response.
Cardiac vagal preganglionic neurons (CVN) control cardiac activity by negative chronotropic, drom... more Cardiac vagal preganglionic neurons (CVN) control cardiac activity by negative chronotropic, dromotropic and inotropic effects. We attempted to characterize the distribution and neuronal properties of the CVN by using double labeling with the retrograde tracer cholera toxin B subunit (CTb) and immunohistochemistry for choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), calcitonin gene-related peptide (CGRP) or nitric oxide synthase (NOS). Injection of CTb into the sinoatrial ganglia resulted in many retrogradely labeled of neurons in the dorsal motor nucleus of the vagus (DMV), the compact (AmC), semicompact (AmS), loose (AmL), external (AmE) formations of the nucleus ambiguus, and the intermediate zone (IZ) between DMV and the nucleus ambiguus. Almost all CTb-labeled neurons showed ChAT immunoreactivity in the DMV, AmC, AmS, AmL and IZ, but most of the CTb-labeled neurons showed no ChAT immunoreactivity in the AmE. Most of the CTb-labeled neurons were double-labeled with CGRP immunoreactivity in the AmC, AmS and AmL, but a few doublelabeled neurons were found in the DMV, IZ and AmE. A few CTb-labeled neurons were double-labeled with NOS immunoreactivity only in the DMV. No TH-immunoreactive neurons were found among the CVN. These results indicate that there are four kinds of neurons among the CVN: non-cholinergic CVN in the AmE, cholinergic and CGRP-containing CVN in the AmC, AmS and AmL, and cholinergic or cholinergic and NOS-containing CVN in the DMV.
The visual cortex in the rat is matured physiologically by postnatal day 30, but the visual syste... more The visual cortex in the rat is matured physiologically by postnatal day 30, but the visual system retains the potential to be reorganized until postnatal day 45. Therefore, we defined the period from postnatal days 28-45 as the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;late critical phase&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;. To examine whether monocular deprivation during the late critical phase gives rise to neuronal apoptosis in the dorsal lateral geniculate nucleus (dLGN), we used the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling method and anterograde tracing. The number of apoptotic cells in the dLGN after monocular deprivation at postnatal day 28 showed little difference from control at postnatal day 29, but was significantly increased to more than fourfold of control ipsilaterally to the monocular deprivation at postnatal day 35 and to more than 10-fold of control bilaterally at postnatal day 40. In the control, there were almost no apoptotic neurons in the dLGN on either side at postnatal day 40. In the nucleus ipsilateral to the monocular deprivation, approximately half the apoptotic neurons were found in an area that did not receive a retinal projection. These findings suggest that the biological process of increased apoptosis in the dLGN of rats that received monocular deprivation in the late critical phase may be different from that in the early critical phase. The increased number of apoptotic cells in the dLGN in the late critical phase may not be simply the result of monocular deprivation.
Biochimica Et Biophysica Acta-proteins and Proteomics, 2011
It is well known that advanced glycation end products (AGEs) are formed in long-lived dermal prot... more It is well known that advanced glycation end products (AGEs) are formed in long-lived dermal proteins such as collagen, and that their formation is related to skin aging. To examine the distribution of AGEs in skin tissue, we performed immunofluorescence studies on the human skin using an anti-AGEs antibody. Interestingly, AGEs signals were observed not only in the dermis but
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