Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 91
Filter
Add more filters










Publication year range
1.
Cell Death Dis ; 4: e700, 2013 Jul 04.
Article in English | MEDLINE | ID: mdl-23828563

ABSTRACT

Math1, a basic helix-loop-helix transcription factor homolog of the Drosophila atonal gene, is considered to be a key factor for induction of sensory hair cells (HCs) during development of the organ of Corti or cochlea. Although embryonic stem (ES) cells are able to produce HC-like cells, the role of Math1 in induction of those cells has not been thoroughly elucidated. In the present study, we introduced Math1 into ES cells in order to achieve efficient generation of HC-like cells. ES cells carrying Tet-inducible Math1, Math1-ES cells, were generated using a Tet-On gene expression system. Embryoid bodies (EBs) formed in the absence of doxycycline (Dox) for 4 days were allowed to grow for an additional 14 days in the dishes in the presence of 400 µg/ml of Dox. At the end of those 14-day cultures, approximately 10% of the cells in EB outgrowths expressed the HC-related markers myosin6, myosin7a, calretinin, α9AchR, and Brn3c (also known as Pou4f3) and showed formation of stereocilia-like structures, whereas few cells in EB outgrowths grown without Dox showed those markers. Reporter assays of Math1-ES cells using a Brn3c-promoter plasmid demonstrated positive regulation of Brn3c by Math1. Furthermore, such HC-related marker-positive cells derived from Math1-ES cells were found to be incorporated in the developing inner ear after transplantation into chick embryos. Math1-ES cells are considered to be an efficient source of ES-derived HC-like cells, and Math1 may be an important factor for induction of HC-like cells from differentiating ES cells.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/physiology , Cell Differentiation , Embryoid Bodies/physiology , Hair Cells, Auditory, Inner/metabolism , Animals , Antigens, Differentiation/metabolism , Cells, Cultured , Chick Embryo , Mechanotransduction, Cellular , Mice , Mice, Transgenic
2.
Neuroscience ; 147(1): 1-4, 2007 Jun 15.
Article in English | MEDLINE | ID: mdl-17521824

ABSTRACT

Fyn, a member of the Src-family protein tyrosine kinase (PTK), is an essential factor in myelination in the CNS and is involved in murine embryonic stem (ES) cell growth and differentiation. Although dysfunctions of Fyn have been comparatively studied, the gain of function by ectopic expression, especially using ES cells, has seldom been investigated. In this article, we give the first report of the involvement of Fyn alteration in the sphere formation ability of murine ES cells. First, transient transfection of Fyn hardly affected multiplication and specialization. Then, we investigated Fyn overexpression using ES cells, which stably express Fyn. As a result, altered sphere formation capability was observed in all clones stably expressing Fyn. These results may provide important information for reproduction medical treatment using ES cells.


Subject(s)
Cell Differentiation/physiology , Embryonic Stem Cells/metabolism , Proto-Oncogene Proteins c-fyn/metabolism , Spheroids, Cellular/metabolism , Animals , Cell Proliferation , Cells, Cultured , Embryonic Stem Cells/cytology , Mice , Spheroids, Cellular/cytology , Transfection
3.
Cell Death Differ ; 14(6): 1080-5, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17318222

ABSTRACT

L3/Lhx8 is a member of the LIM-homeobox gene family. Previously, we demonstrated that L3/Lhx8-null mice specifically lacked cholinergic neurons in the basal forebrain. In the present study, we conditionally suppressed L3/Lhx8 function during retinoic acid-induced neural differentiation of a murine embryonic stem (ES) cell line using an L3/Lhx8-targeted small interfering RNA (siRNA) produced by an H1.2 promoter-driven vector. Our culture conditions induced efficient differentiation of the ES cells into neurons and astrocytes, but far less efficient differentiation into oligodendrocytes. Suppression of L3/Lhx8 expression by siRNA led to a dramatic decrease in the number of cells positive for the cholinergic marker ChAT, and overexpression of L3/Lhx8 recovered this effect. However, no significant changes were observed in the number of Tuj1+ neurons and GABA+ cells. These results strongly suggest that L3/Lhx8 is a key factor in the cholinergic differentiation of murine ES cells and is involved in basal forebrain development.


Subject(s)
Cell Differentiation/physiology , Embryonic Stem Cells/metabolism , Homeodomain Proteins/physiology , Animals , Astrocytes/cytology , Astrocytes/metabolism , Cell Differentiation/genetics , Cells, Cultured , Choline O-Acetyltransferase/genetics , Choline O-Acetyltransferase/metabolism , Embryonic Stem Cells/cytology , Gene Expression , Genetic Vectors/genetics , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Immunohistochemistry , In Situ Nick-End Labeling , LIM-Homeodomain Proteins , Mice , Mutation , Neurons/cytology , Neurons/metabolism , Oligodendroglia/cytology , Oligodendroglia/metabolism , Prosencephalon/cytology , Prosencephalon/metabolism , RNA, Small Interfering/genetics , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factors , Transfection
4.
J Neurochem ; 94(3): 723-30, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16000160

ABSTRACT

The LIM homeobox family of transcription factors is involved in many processes during the development of the mammalian central nerves system. L3, also called Lhx8 (L3/Lhx8), is a recently identified member of the LIM homeobox gene family and is selectively expressed in the medial ganglionic eminence (MGE). Our previous study demonstrated that L3/Lhx8-null mice specifically lacked cholinergic neurons in the basal forebrain. In this study, we reduced L3/Lhx8 function in the murine neuroblastoma cell line, Neuro2a (N2a), using L3/Lhx8-targeted small interfering RNA (siRNA) produced by H1.2 promoter-driven vector. The levels of cholinergic markers per cell were diminished without a reduction in the number of marker-positive cells. Intriguingly, GABAergic marker expression and the number of GABAergic cells were dramatically increased in the differentiating L3/Lhx8-knockdown N2a. These results suggest the possibility that L3/Lhx8 is involved in the determination of transmitter phenotypes (GABAergic or cholinergic cell fate) in a population of neurons during basal forebrain development.


Subject(s)
Cell Differentiation/drug effects , Choline O-Acetyltransferase/metabolism , Homeodomain Proteins/metabolism , RNA, Small Interfering/pharmacology , gamma-Aminobutyric Acid/metabolism , Animals , Blotting, Northern/methods , Cell Count/methods , Cell Death/drug effects , Cell Differentiation/physiology , Cell Line, Tumor , Cyclic AMP/pharmacology , Drug Interactions , Gene Expression Regulation/drug effects , Genetic Vectors/physiology , Glutamate Decarboxylase/metabolism , Green Fluorescent Proteins/biosynthesis , Homeodomain Proteins/genetics , Homeodomain Proteins/pharmacology , In Situ Nick-End Labeling/methods , Isoenzymes/metabolism , Ki-67 Antigen/metabolism , LIM-Homeodomain Proteins , Membrane Transport Proteins/genetics , Membrane Transport Proteins/metabolism , Mice , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neuroblastoma , RNA, Messenger/metabolism , RNA, Small Interfering/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction/methods , Time Factors , Transcription Factors , Transfection/methods , Tretinoin/pharmacology , Vesicular Acetylcholine Transport Proteins
5.
Orthod Craniofac Res ; 5(2): 65-70, 2002 May.
Article in English | MEDLINE | ID: mdl-12086327

ABSTRACT

OBJECTIVES: To compare and contrast the gene expression of two LIM-homeobox type transcription factors, Lhx6 and L3/Lhx8, in secondary palate formation. METHODS: In situ hybridization histochemistry with digoxygenin (DIG) labelled cRNA probes specific for Lhx6 and L3/Lhx8. MATERIALS: Serial cryo-sections of embryonic day (E)13.5, 14.5, and 15.5 mice (C57BL/6). OUTCOME MEASURE: Comparison of the signal intensities of NBT/BCIP precipitate by alkaline phosphatase conjugated anti-DIG antibody. RESULTS: From E13.5 to E15.5, Lhx6 and L3/Lhx8 signals are detected in palatal mesenchyme, but the L3/Lhx8 signal is much more intense than the Lhx6 signal. In palatal epithelium, covering the mesenchyme, Lhx6 mRNA is transiently expressed at E14.5, while L3/Lhx8 mRNA expression is never detected throughout the development. CONCLUSION: Lhx6 and L3/Lhx8 functions may be partially redundant in the mesenchyme of the secondary palate, but not in the palatal epithelium.


Subject(s)
Cleft Palate/embryology , Homeodomain Proteins/biosynthesis , Nerve Tissue Proteins , Palate, Hard/embryology , Animals , Embryonic and Fetal Development , Epithelium/embryology , Epithelium/metabolism , Gene Expression Regulation, Developmental , Genes, Homeobox/physiology , LIM-Homeodomain Proteins , Mesoderm/metabolism , Mice , Mice, Inbred C57BL , RNA Probes , Transcription Factors
6.
Bone ; 29(5): 458-66, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11704499

ABSTRACT

Recent experiments have shown that exogenous basic fibroblast growth factor (bFGF) enlarges fracture callus and accelerates the healing of osteotomized long bones. The actions of bFGF are mediated by four different transmembrane receptors (FGFR1-4). Among them, FGFR1 has a high affinity for bFGF, and gain-of-function mutations of the FGFR1 gene cause craniosynostosis in humans. Gene expression for FGFR1 has been analyzed in embryogenesis; however, in skeletal repair, detailed expression of FGFR1 has not been fully established. In the present study, a rat model of closed femoral fracture healing was used to quantify mRNA encoding the FGFR1 and to characterize cells expressing FGFR1 by in situ hybridization. Gene expression for FGFR1 was rapidly upregulated after fracture; its mRNA level on day 1 was 3.4-fold higher than that of unfractured femora. At this stage, a moderate signal for FGFR1 was detected in periosteal osteoprogenitor cells, inflammatory cells near fracture sites, and cells among muscle layers. FGFR1 mRNA reached peak expression when callus remodeling actively progressed (6.8-fold on day 14), and remained elevated even in the later stages of healing (6.3-fold on day 28). During the intermediate stage of fracture healing, a strong signal for FGFR1 was diffusely distributed in mature osteoblasts in the hard callus, and mature osteoclasts also expressed a weak signal for FGFR1. These results suggest that FGF/FGFR1 signaling has multifunctional roles during fracture healing and may regulate both osteoblasts and osteoclasts, contributing to bone formation and callus remodeling.


Subject(s)
Fracture Healing/physiology , Receptor Protein-Tyrosine Kinases/genetics , Receptors, Fibroblast Growth Factor/genetics , Animals , Bony Callus/physiopathology , Femur/injuries , Femur/physiology , Fibroblast Growth Factor 2/genetics , Gene Expression/physiology , In Situ Hybridization , Male , RNA, Messenger/analysis , Rats , Rats, Sprague-Dawley , Receptor, Fibroblast Growth Factor, Type 1 , Ribonucleases , Signal Transduction/physiology
7.
Neurosci Lett ; 313(3): 153-7, 2001 Nov 09.
Article in English | MEDLINE | ID: mdl-11682150

ABSTRACT

DNA fragmentation and activation of caspase-1, implicating involvement of apoptosis, have been reported in the spinal cord of amyotrophic lateral sclerosis (ALS) patients and transgenic mouse models of ALS. Because BH3-only members of the Bcl-2 family have pro-apoptotic activity, we examined the expression of the BH3-only peptide harakiri (Hrk) in the spinal cord of ALS patients. In situ expression of Hrk mRNA and immunoreactivity against the Hrk peptide were verified in the spinal neurons. In the immunoblot analysis, upregulated Hrk protein migrated at 16 kDa. Heterodimerization of Hrk with Bcl-2 was detected by immunoprecipitation, which suggests the competition of Hrk and anti-apoptotic Bcl-2. These findings suggest that Hrk plays a role in apoptotic events in ALS pathogenesis.


Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Carrier Proteins/metabolism , Neurons/metabolism , Spinal Cord/metabolism , Up-Regulation , Adult , Aged , Amyotrophic Lateral Sclerosis/pathology , Apoptosis , Carrier Proteins/genetics , Female , Humans , Immunoblotting , Immunohistochemistry , In Situ Hybridization , In Situ Nick-End Labeling , Male , Middle Aged , Neurons/pathology , Precipitin Tests , Protein Binding , Protein Structure, Tertiary/physiology , Proto-Oncogene Proteins c-bcl-2/metabolism , RNA, Messenger/biosynthesis , Spinal Cord/pathology
8.
Histochem Cell Biol ; 116(2): 141-8, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11685542

ABSTRACT

The OASIS gene, which encodes a novel CREB/ATF family member, was isolated from long-term cultured astrocytes that were employed as an in vitro gliosis model. In the present study, we examined the expression pattern of the OASIS gene in the developing mouse embryo by in situ hybridization histochemistry and compared it with the expression of osteogenesis markers. OASIS mRNA expression was most strongly detected in preosteoblasts of the outer bony cortex of the ribs. Alveolar bone also showed strong signals for OASIS gene expression. OASIS mRNA was also localized to the preodontoblast of tooth buds. Expression began at embryonic day 12 (D12.5), peaked around D14.5-16.5, and continued to D18.5. The pattern of expression was very similar to that of hXBP-1 mRNA, which encodes another CREB/ATF family member. Spatiotemporal patterns of OASIS partly overlapped that of osteopontin, osteonectin, and alpha1 type I procollagen genes. Among these, the time course of OASIS mRNA expression was most similar to that of osteopontin mRNA expression, suggesting that the OASIS protein is involved in the late phase of osteoblast differentiation, as compared to the Cbfa1 that regulates early phases of osteoblast differentiation.


Subject(s)
Bone Development/physiology , Gene Expression Regulation, Developmental , Nerve Tissue Proteins , Transcription Factors/genetics , Animals , Cartilage/embryology , Cartilage/physiology , Collagen/genetics , Cyclic AMP Response Element-Binding Protein , DNA-Binding Proteins/genetics , Exocrine Glands/embryology , Exocrine Glands/physiology , Female , Fetus/physiology , In Situ Hybridization , Mice , Mice, Inbred ICR , Odontoblasts/physiology , Osteoblasts/physiology , Osteonectin/genetics , Osteopontin , Pregnancy , RNA, Messenger/analysis , Regulatory Factor X Transcription Factors , Ribs/cytology , Ribs/embryology , Ribs/physiology , Sialoglycoproteins/genetics , Tooth Germ/cytology , Tooth Germ/embryology , Tooth Germ/physiology
9.
Cell Immunol ; 204(1): 11-8, 2000 Aug 25.
Article in English | MEDLINE | ID: mdl-11006013

ABSTRACT

Endogenous retroviral gene products have been recognized as being expressed in human cancerous tissues. However, these products have not been shown to be antigenic targets for T-cells, possibly due to immune tolerance. Since carcinogen-induced colon tumor CT26 expresses an envelope protein, gp70, of an endogenous ecotropic murine leukemia virus that is comparable to human tumor-associated antigens, we examined whether a DNA vaccine containing the gp70 gene induces protective immunity against CT26 cells. Injection of mice with plasmid DNA (pDNA) encoding gp70 alone failed to induce anti-gp70 antibody (Ab) or anti-CT26 cytotoxic T lymphocyte (CTL) responses. However, immunization with pDNA encoding the beta-galactosidase (beta-gal)/gp70 fusion protein induced anti-gp70 Ab and anti-CT26 CTL responses and conferred protective immunity against CT26 cells. These results indicate that beta-gal acts as an immunogenic carrier protein that helps in the induction of immune responses against the poorly immunogenic gp70. Considering these results, it is possible that potential tolerance to the endogenous retroviral gene products expressed by human tumors may be overcome by DNA vaccines that contain an endogenous retroviral gene fused to genes encoding immunogenic carrier proteins.


Subject(s)
Cancer Vaccines/therapeutic use , Colonic Neoplasms/therapy , Leukemia Virus, Murine/immunology , Retroviridae Proteins, Oncogenic/immunology , Vaccines, DNA/therapeutic use , Viral Envelope Proteins/immunology , Animals , Antibodies, Neoplasm/blood , Colonic Neoplasms/mortality , Cytotoxicity, Immunologic , Female , Leukemia Virus, Murine/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Neoplasm Proteins/immunology , Recombinant Fusion Proteins/immunology , Retroviridae Proteins, Oncogenic/genetics , T-Lymphocytes, Cytotoxic/immunology , Vaccination , Viral Envelope Proteins/genetics , beta-Galactosidase/genetics , beta-Galactosidase/immunology
11.
J Chem Neuroanat ; 17(2): 99-107, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10585161

ABSTRACT

Huntingtin-interacting protein-2 (Hip-2) was identified as a human protein specifically associated with huntingtin in vitro, a gene product affected in patients with Huntington disease (HD). It is a ubiquitin-conjugating enzyme identical to the previously characterized bovine E2-25k. We identified the mouse Hip-2 homologue (mHip-2) and examined its distribution patterns in the developing mouse brain in order to gain an insight into the functional significance of the Hip-2 protein in the normal brain as well as in the pathogenesis of HD. As reported with huntingtin, the mHip-2 mRNA expression developed in parallel with neuronal maturation and became distributed widely in the postnatal mouse brain. This spatiotemporal pattern of mHip-2 mRNA expression resembled that of huntingtin. We further demonstrated that mHip-2 mRNA was colocalized with huntingtin-like immunoreactivity in a single neuron. These findings suggested that the Hip-2 interacted with huntingtin in vivo and played an important role in HD pathogenesis.


Subject(s)
Brain/metabolism , Ligases/genetics , Ubiquitin-Conjugating Enzymes , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Brain/anatomy & histology , Brain/growth & development , DNA, Complementary/analysis , Huntingtin Protein , In Situ Hybridization , Ligases/chemistry , Ligases/metabolism , Mice , Mice, Inbred ICR , Molecular Sequence Data , Nerve Tissue Proteins/metabolism , Nuclear Proteins/metabolism , RNA, Messenger/metabolism
12.
Pathol Int ; 49(8): 687-94, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10504535

ABSTRACT

In basal cell nevus syndrome (BCNS) patients, mutations of a gene, patched (ptc), which encodes a putative signal transducer of sonic hedgehog protein (SHH), were found and are thought to be one of the major causes of BCNS. The SHH signaling pathway is an important developmental pathway, and ptc protein (PTC) is a suppressive component serving as a receptor for the secreted SHH. Another transmembrane protein, smoothened (SMO), forms a complex with PTC and regulates this signaling pathway. Recent transgenic studies have strengthened the importance of the SHH signaling system in the etiology of basal cell carcinoma (BCC). In this study, we examined the expression patterns of mRNA for ptc and smo in two different BCC subtypes and normal skin. We found that the expressions of ptc and smo mRNA were enhanced in the tumor nests of the nodular BCC, especially at the advancing portions, but were under the detectable level in the superficial BCC cases examined, indicating that ptc and smo mRNA expressions might be associated with BCC tumor progression and divide the BCC histologic types into two subtypes, superficial and nodular types. In addition, no obvious signals for ptc and smo mRNA were detected in the normal human epidermis, appendages, or seborrheic keratosis, indicating that the abnormal proliferation of follicular epithelial cells caused by ptc, smo and/or other genetic changes, which also cause ptc and smo overexpressions, might result in BCC tumor formation.


Subject(s)
Carcinoma, Basal Cell/genetics , Gene Expression , Membrane Proteins/genetics , Proteins/physiology , Receptors, Cell Surface/genetics , Receptors, G-Protein-Coupled , Signal Transduction , Skin Neoplasms/genetics , Trans-Activators , Aged , Aged, 80 and over , Hedgehog Proteins , Humans , In Situ Hybridization , Middle Aged , Patched Receptors , Patched-1 Receptor , RNA, Messenger/metabolism , Smoothened Receptor
13.
Brain Res Mol Brain Res ; 69(1): 93-103, 1999 May 21.
Article in English | MEDLINE | ID: mdl-10350641

ABSTRACT

Gliosis is a characteristic response of astrocytes to inflammation and trauma of the central nervous system (CNS). To study the mechanisms underlying gliosis, we performed differential display screening for genes specifically induced in long-term cultured astrocytes used as an in vitro gliosis model. We identified and characterized a gene (named OASIS, for old astrocyte specifically-induced substance) expressed in long-term cultured mouse astrocytes, or 'old astrocytes (OA)'. The OASIS gene encoded a putative transcription factor belonging to the cyclic AMP responsive element binding protein/activating transcription factor (CREB/ATF) gene family, with homology to box B-binding factor-2 (BBF-2), a Drosophila transcription factor. Its expression was developmentally regulated; OASIS mRNA was primarily expressed in the salivary gland and cartilage in the mouse embryo and it was transiently upregulated in the brain during postnatal two weeks. The expression became weaker in the adult brain. We also demonstrated that an expression of the OASIS mRNA was induced in response to the cryo-injury of the mouse cerebral cortex. The distribution pattern of the OASIS-positive cells in the injured cortex was very similar to that of the glial fibrillary acidic protein (GFAP)-positive cells. These results suggest that OASIS protein may play a role in gliotic events.


Subject(s)
Astrocytes/chemistry , Astrocytes/physiology , Blood Proteins/genetics , Cyclic AMP Response Element-Binding Protein/genetics , Nerve Tissue Proteins , Transcription Factors/genetics , Activating Transcription Factors , Age Factors , Animals , Blotting, Northern , Cells, Cultured , Cerebral Cortex/cytology , Cloning, Molecular , Glial Fibrillary Acidic Protein/analysis , Gliosis/immunology , In Situ Hybridization , Male , Mice , Mice, Inbred ICR , Molecular Sequence Data , Neuritis/genetics , Neuritis/immunology , RNA, Messenger/analysis , Sequence Homology, Amino Acid
14.
FEBS Lett ; 447(1): 76-80, 1999 Mar 19.
Article in English | MEDLINE | ID: mdl-10218586

ABSTRACT

We identified a novel member of the Ikaros gene family, which has critical roles in the development of lymphoid lineages. This gene, which we named Eos, was expressed predominantly in the developing central and peripheral nervous system. Eos protein could interact with itself and Ikaros protein through its C-terminal portion in the yeast two hybrid assay. These findings suggested that Eos may have important roles in neural development similarly to the Ikaros family in the development of hemolymphoid tissue.


Subject(s)
Carrier Proteins/genetics , DNA-Binding Proteins , Multigene Family , Nerve Tissue Proteins/genetics , Zinc Fingers/genetics , Amino Acid Sequence , Animals , Astrocytes/cytology , Carrier Proteins/isolation & purification , Central Nervous System/chemistry , Ikaros Transcription Factor , In Situ Hybridization , Mice , Mice, Inbred ICR , Molecular Sequence Data , Nerve Tissue Proteins/isolation & purification , Peripheral Nervous System/chemistry , Protein Binding , RNA, Messenger/isolation & purification , Sequence Homology, Amino Acid , Tissue Distribution , Transcription Factors/genetics , Transcription Factors/metabolism
15.
J Biol Chem ; 274(12): 7975-81, 1999 Mar 19.
Article in English | MEDLINE | ID: mdl-10075695

ABSTRACT

DP5, which contains a BH3 domain, was cloned as a neuronal apoptosis-inducing gene. To confirm that DP5 interacts with members of the Bcl-2 family, 293T cells were transiently co-transfected with DP5 and Bcl-xl cDNA constructs, and immunoprecipitation was carried out. The 30-kDa Bcl-xl was co-immunoprecipitated with Myc-tagged DP5, suggesting that DP5 physically interacts with Bcl-xl in mammalian cells. Previously, we reported that DP5 is induced during neuronal apoptosis in cultured sympathetic neurons. Here, we analyzed DP5 gene expression and the specific interaction of DP5 with Bcl-xl during neuronal death induced by amyloid-beta protein (A beta). DP5 mRNA was induced 6 h after treatment with A beta in cultured rat cortical neurons. The protein encoded by DP5 mRNA showed a specific interaction with Bcl-xl. Induction of DP5 gene expression was blocked by nifedipine, an inhibitor of L-type voltage-dependent calcium channels, and dantrolene, an inhibitor of calcium release from the endoplasmic reticulum. These results suggested that the induction of DP5 mRNA occurs downstream of the increase in cytosolic calcium concentration caused by A beta. Moreover, DP5 specifically interacts with Bcl-xl during neuronal apoptosis following exposure to A beta, and its binding could impair the survival-promoting activities of Bcl-xl. Thus, the induction of DP5 mRNA and the interaction of DP5 and Bcl-xl could play significant roles in neuronal degeneration following exposure to A beta.


Subject(s)
Amyloid beta-Peptides/pharmacology , Apoptosis , Gene Expression Regulation , Neurons/physiology , Neuropeptides/biosynthesis , Neuropeptides/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Alzheimer Disease/physiopathology , Amino Acid Sequence , Animals , Apoptosis/genetics , Apoptosis Regulatory Proteins , Cells, Cultured , Molecular Sequence Data , Protein Binding , RNA, Messenger/metabolism , Rats
16.
Brain Res Mol Brain Res ; 62(2): 220-3, 1998 Nov 20.
Article in English | MEDLINE | ID: mdl-9813338

ABSTRACT

Tra2beta is the first mammalian protein which is proved to activate mRNA splicing in sequence-specific manner. Following hypoglossal nerve injury, the expression of Tra2beta mRNA was elevated in injured motoneurons transiently. The up-regulation of Tra2beta mRNA was observed from post-operative day 3 to 21. In addition to the nerve injury in PNS, a brain lesion in CNS also enhanced the expression of Tra2beta mRNA. The present study could be the first observation showing that an expression of the sequence-specific splicing activator is enhanced in neuronal cells in response to nerve injury, and indicates that Tra2beta may participate in the control of injury-specific splicing patterns in order to express molecules which are necessary for regeneration.


Subject(s)
Gene Expression Regulation , Hypoglossal Nerve Injuries , Medulla Oblongata/metabolism , Nerve Regeneration/genetics , Nerve Tissue Proteins/biosynthesis , RNA Splicing , RNA-Binding Proteins/biosynthesis , Temporal Lobe/metabolism , Animals , Axotomy , Dominance, Cerebral , Hypoglossal Nerve/metabolism , Male , Motor Neurons/metabolism , Nerve Tissue Proteins/genetics , RNA-Binding Proteins/genetics , Rats , Rats, Wistar , Serine-Arginine Splicing Factors
17.
Brain Res Mol Brain Res ; 61(1-2): 179-89, 1998 Oct 30.
Article in English | MEDLINE | ID: mdl-9795207

ABSTRACT

In the course of a study conducted to identify the mouse homologue of Drosophila eyes absent (eya), we isolated a novel mouse cDNA fragment which show little homology to eya but encodes a protein with Krüppel (C2H2)-type zinc finger motifs. By further screening using this cDNA fragment as a probe, we obtained the short and long forms of full-length cDNAs, which were apparently alternatively spliced products from one gene. Since both mRNAs encode proteins with widely-interspaced zinc finger motifs, we termed this gene wiz and refer to the short and long wiz transcripts as wizS and wizL, respectively. In situ hybridization studies using the probe against the region common to wizS and wizL showed that these mRNAs were expressed abundantly in the granule cell layers of the mouse cerebellum, the olfactory bulb, and the dentate gyrus, whereas the same technique using the probe against only wizL could not detect positive signals in the developing cerebellum, indicating that there is no expression of wizL mRNA there. Northern blot and in situ hybridization analyses demonstrated that the extracerebellar regions expressed both wizS and wizL mRNAs from the midgestational period to adulthood. The finding that two types of wiz transcripts (wizS and wizL) are expressed with different developmental patterns might indicate separate transcription functions in the cerebellar granule cells and the extracerebellar regions.


Subject(s)
Alternative Splicing/genetics , Cerebellum/chemistry , Gene Expression Regulation, Developmental/genetics , Nerve Tissue Proteins/genetics , Transcription Factors/genetics , Zinc Fingers/genetics , Amino Acid Sequence , Animals , Animals, Newborn , Blotting, Northern , Brain , Cerebellum/cytology , Cerebellum/metabolism , Cloning, Molecular , Kruppel-Like Transcription Factors , Mice , Mice, Inbred ICR , Molecular Sequence Data , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/isolation & purification , Organ Specificity/genetics , RNA, Messenger/biosynthesis , Transcription Factors/biosynthesis , Transcription Factors/isolation & purification
19.
Brain Res Mol Brain Res ; 54(2): 316-20, 1998 Mar 01.
Article in English | MEDLINE | ID: mdl-9555074

ABSTRACT

We examined the expression patterns of the DP5 gene, which encodes a protein with apoptosis-inducing activity, in the developing nervous system of mice. This gene was primarily expressed in the spinal motor neurons and peripheral sensory ganglia of mouse embryos and transiently in the postnatal brain, particularly in the entorhinal cortex and hippocampus. These expression patterns suggest that the DP5 gene may be involved in the apoptosis, if not all, of the developing nervous system.


Subject(s)
Gene Expression Regulation, Developmental/physiology , Nervous System/metabolism , Amino Acid Sequence , Animals , Apoptosis/genetics , Ganglia, Sensory/metabolism , In Situ Hybridization , Mice , Mice, Inbred ICR , Molecular Sequence Data , Motor Neurons/metabolism , Nervous System/embryology , Nervous System/growth & development
20.
Genomics ; 49(2): 307-9, 1998 Apr 15.
Article in English | MEDLINE | ID: mdl-9598319

ABSTRACT

Lhx8 is a LIM-homeodomain protein, containing two tandemly repeated LIM motifs and a hemeodomain. The expression of Lhx8 is limited spatially to the medical ganglionic eminence and the mesenchyme surrounding the oral cavity and temporally from middle embryonic to early postnatal development, suggesting a role for Lhx8 in differentiation of certain neurons and mesenchymal cells, just as the other LIM-homeodomain proteins are implicated in determining the fates of certain cell types. Here we report the structure and the chromosomal localization of the Lhx8 gene. The gene is composed of nine exons and eight introns. The first LIM domain is coded by two exons, exons 2 and 3, and the second by a single exon, exon 4. The homeodomain is encoded by three exons, exons 6, 7, and 8. In situ chromosomal hybridization demonstrated that the Lhx8 gene was localized in the distal region of mouse chromosome 3.


Subject(s)
Chromosome Mapping , Genes, Homeobox , Homeodomain Proteins/genetics , Transcription Factors/genetics , Animals , Embryo, Mammalian , Exons/genetics , In Situ Hybridization , LIM-Homeodomain Proteins , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Sequence Analysis, DNA
SELECTION OF CITATIONS
SEARCH DETAIL
...