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At least 649 records · Page 36Linked to original sources
PubMed1996

Cloning, characterization and transient expression of the gene encoding a rice U3 small nuclear RNA.

A rice U3 small nuclear RNA (snRNA)-encoding gene has been isolated. The coding region of this gene contains all five conserved sequence boxes common to plant U3 snRNAs. The upstream and downstream regions of the gene harbour characteristic sequence elements required for transcription with RNA polymerase III (pol III), as well as three monocot-specific promoter (MSP) elements [Connelly et al., Mol. Cell Biol.14 (1994) 5910-5919], two of them comprising a palindromic G+C-rich segment. The sequence (TTTAAAA) of the TATA-box in this gene does not fit the established consensus [Marshallsay et al., Plant Mol. Biol. 19(1992) 973-983], making this gene unique among reported snRNA-encoding genes of plants. An RNase protection assay showed that the gene is expressed properly in cucumber protoplasts. We thus suggest that, with other promoter elements present, the TATA-box for RNA pol III-specific snRNA-encoding genes may not be as well conserved as that for RNA pol II-specific genes.

Base Sequence↗
PubMed1996

The receptor tyrosine kinase, Cek8, is transiently expressed on subtypes of motoneurons in the spinal cord during development.

Receptor tyrosine kinases (RTKs) play important roles in cellular proliferation, differentiation, and survival. We performed reverse transcriptase-polymerase chain reactions (RT-PCR) from enriched embryonic day 5 (E5) chick motoneurons by panning to identify RTKs involved in the early development of motoneuron. In situ hybridization revealed that Cek8, a member of the eph family, was specifically expressed on motoneurons at the brachial and lumbar segments of the spinal cord which innervate limb muscles, and disappeared after the naturally occurring cell death period (E6-E11). Immunohistochemistry using an anti-Cek8 monoclonal antibody showed the localization of Cek8 protein at the cell bodies and axonal fibers of motoneurons and muscles. The unique expression of Cek8 suggests its involvement in cellular survival or cell-cell interactions for specific subpopulations of developing motoneurons.

Amino Acid Sequence↗
PubMed1993

Current applications of COS cell based transient expression systems.

An ever increasing number of mammalian expression systems have become available in recent years. Yet a simple and robust mammalian expression system is all that is needed for most routine mammalian expression work. The well established COS cell based expression systems have filled this role and continue to be used to study gene expression, to clone by expression, to produce small quantities of recombinant protein, and to test the efficacy of mammalian expression constructs. Recent applications of COS cell based expression systems in these areas include the following: studies on the role of AU-rich regions localized in the 3' untranslated regions of mRNA transcripts in mRNA half-life; the cloning of the leukocyte antigens CD34 and CD69 as well as the type-II and type-III TGF-beta receptors; and the production of a soluble recombinant form of the gamma delta T-cell receptor and a bispecific Ig fusion protein of the endothelial cell-surface proteins E-selectin and P-selectin.

Animals↗
PubMed2005

MN-cadherin and its novel variant are transiently expressed in chick embryo spinal cord.

To isolate cDNAs that are involved in limb-motoneuron development, we compared mRNAs of lumbar and thoracic motoneurons purified from spinal cord of E4 chick embryo by differential display. In situ hybridization demonstrated that one of cDNAs is expressed exclusively in lateral motor column in spinal cord from E4 to E10. We identified two mRNA variants for the cDNA by library screening. The long form (788 amino acids) was identical to chick MN-cadherin. The short variant (543 amino acids) lacks the first two of five extracellular domains of MN-cadherin, which commonly exist in classical cadherins. The amino acid sequence of the short form is identical to that of the carboxyl terminal MN-cadherin, except for the distinct signal sequence. The ratio of mRNA of short form to long form was 1-20. cDNA transfection study revealed that the long form but not the short form MN-cadherin had cell adhesion activity.

Amino Acid Sequence↗
PubMed2004

Transient expression of GABAA receptor alpha2 and alpha3 subunits in differentiating cerebellar neurons.

In the adult mammalian brain, synaptic transmission mediated by gamma-amino butyric acid (GABA) plays a role in inhibition of excitatory synaptic transmission. During brain development, GABA is involved in brain morphogenesis. To clarify how GABA exerts its effect on immature neurons, we examined the expression of the GABAA receptor alpha2 and alpha3 subunits, which are abundantly expressed before alpha1 and alpha6 subunits appear, in the developing mouse cerebellum using in situ hybridization. Proliferating neuronal precursors in the ventricular zone and external granular layer expressed neither alpha2 nor alpha3 subunits. Hybridization signals for the alpha2 and alpha3 subunit mRNAs first appeared in the differentiating zone at embryonic day 13 (E13). The alpha2 subunit was detected in the migrating and differentiating granule cells and cerebellar nucleus neurons until postnatal day 14 (P14). Hybridization signals for the alpha3 subunit mRNA, on the other hand, were localized in the developing Purkinje cells and cerebellar nucleus neurons, and disappeared from Purkinje cells by the end of first postnatal week. Taken together, this indicated that the alpha2 and alpha3 subunits were abundantly expressed in distinct types of cerebellar neurons after completing cell proliferation while forming the neural network. These results suggest that GABA might extrasynaptically activate the GABAA receptors containing alpha2 and/or alpha3 subunits on the differentiating neurons before finishing the formation of synapses and networks, and could be involved in neuronal differentiation and maturation in the cerebellum.

Animals↗
PubMed2006

Reelin is transiently expressed in the peripheral nerve during development and is upregulated following nerve crush.

Reelin is an extracellular matrix protein which is critical for the positioning of migrating post-mitotic neurons and the laminar organization of several brain structures during development. We investigated the expression and localization of Reelin in the rodent peripheral nerve during postnatal development and following crush injury in the adult stage. As shown with Western blotting, immunocytochemistry and RT-PCR, Schwann cells in the developing peripheral nerve and in primary cultures from neonatal nerves produce and secrete Reelin. While Reelin levels are downregulated in adult stages, they are again induced following sciatic nerve injury. A morphometric analysis of sciatic nerve sections of reeler mice suggests that Reelin is not essential for axonal ensheathment by Schwann cells, however, it influences the caliber of myelinated axons and the absolute number of fibers per unit area. This indicates that Reelin may play a role in peripheral nervous system development and repair by regulating Schwann cell-axon interactions.

Animals↗
PubMed2003

Thalamocortical cells in the cat pulvinar nucleus transiently express nitric oxide synthase during development.

We examined the postnatal expression of the neuronal form of nitric oxide synthase (nNOS) within the pulvinar and lateral posterior (LP) nuclei of the cat thalamus using immunocytochemical techniques. During the first postnatal month, nNOS was expressed in many cells within the pulvinar nucleus and medial subdivision of the LP nucleus; fewer neurons in the lateral LP nucleus were stained by the nNOS antibody. We examined the pulvinar nucleus to determine what cell types express nNOS. A comparison of the soma sizes of nNOS-stained cells to the overall population of Nissl-stained cells and interneurons (stained with an antibody against glutamic acid decarboxylase) suggests that within the pulvinar nucleus, thalamocortical cells express nNOS during development. In addition, the nNOS antibody stained axon bundles that traverse the pulvinar nucleus to enter the optic radiations, suggesting that thalamocortical cell axons also contain nNOS during development. However, this staining pattern was dramatically reduced by postnatal day 42 and later ages; the size of the remaining nNOS-stained cells was closer to that of interneurons, a subset of which contain nNOS in the adult pulvinar nucleus. This contrasts with our previous findings that nNOS is specifically expressed within interneurons in the developing dorsal lateral geniculate nucleus (LGN) and serves as further confirmation that the pulvinar nucleus and LGN represent distinct categories of thalamic nuclei.

Animals↗
PubMed2006

Transient expression of Human papillomavirus type 16 L1 protein in Nicotiana benthamiana using an infectious tobamovirus vector.

A Tobacco mosaic virus (TMV)-derived vector was used to express a native Human papillomavirus type 16 (HPV-16) L1 gene in Nicotiana benthamiana by means of infectious in vitro RNA transcripts inoculated onto N. benthamiana plants. HPV-16 L1 protein expression was quantitated by enzyme-linked immunosorbent assays (ELISA) after concentration of the plant extract. We estimated that the L1 product yield was 20-37 microg/kg of fresh leaf material. The L1 protein in the concentrated extract was antigenically characterised using the neutralising and conformation-specific Mabs H16:V5 and H16:E70, which bound to the plant-produced protein. Particles observed by transmission electron microscopy were mainly capsomers but virus-like particles (VLPs) similar to those produced in other systems were also present. Immunisation of rabbits with the concentrated plant extract induced a weak immune response. This is the first report of the successful expression of an HPV L1 gene in plants using a plant virus vector.

Antibodies, Monoclonal↗
PubMed1998

Muscle/heart isoform of mitochondrial adenine nucleotide translocase (ANT1) is transiently expressed during perinatal development in rat liver.

A postnatal increase in the content of mitochondrial ANT in rat liver which is related to the maturation of mitochondrial function has previously been reported [Schönfeld et al., Biochim. Biophys Acta 1144 (1993) 353-358]. In order to define the contribution of the ANT isoforms to this postnatal increase we have studied the expression of ANT1 and ANT2 isoforms in the liver during this period. The results show that in contrast to adult liver, perinatal liver expressed the ANT1 isoform at the mRNA and protein level, and that during this period the expression of ANT1 increased to a similar extent as total ANT content. It is concluded that the postnatal increase in ANT is mainly due to the ANT1 isoform and therefore, a role for the ANT1 isoform in the postnatal maturation of mitochondrial respiration in rat liver is suggested.

Animals↗
PubMed1999

Transient expression assays with the proximal promoter of a newly characterized actin gene from the oyster Crassostrea gigas.

We undertook the characterization of an actin gene and its proximal promoter in the oyster Crassostrea gigas. A complete actin cDNA was identified, sequenced and its amino acid sequence deduced. Comparative analysis showed a high homology with actin of other species and that this gene is closer to the cytoplasmic form of actins than to the muscle type. A probe derived from the 5'-untranslated region of the cDNA was then used to isolate the actin gene from a genomic library. The gene was sequenced and shown to contain a single 643 bp intron. A 1670 bp fragment upstream from the open reading frame was isolated and sequenced. This upstream region displays typical features of actins such as a serum response element (CarG box). This fragment was cloned into the promoterless vector pGL3-basic and the resulting construct was transfected into cells of dissociated oyster heart primary cultures. Its capacity to express the luciferase in this in vitro homologous system was monitored and showed high expression levels. This is the first complete actin sequence reported so far for the oyster C. gigas and its promoter is the first available among bivalves.

Actins↗
PubMed1988

An embryonic-like myosin heavy chain is transiently expressed in nodal conduction tissue of the rat heart.

In the bovine nodal conduction tissue we have described the existence of a novel cardiac myosin isoform, immunologically related to the myosin types expressed during skeletal muscle development. Using different monoclonal antibodies specific for the embryonic and the neonatal skeletal myosin heavy chain types we investigated the myosin composition of the rat sino-atrial and atrio-ventricular nodes. We find that nodal conduction tissue fibers of the rat heart contain a distinct cardiac myosin isoform antigenically similar to the skeletal embryonic myosin heavy chain. The expression of this myosin isoform in nodal tissue appears to be developmentally regulated and partially controlled by thyroid hormone. Reactive cardiac fibers were detected in the nodal regions only during fetal development and a few days after birth, whereas very rare labelled fibers could be observed in the adult nodes. This myosin type does not represent a primordial cardiac myosin isoform since it was not detected in the embryonic heart before 13.5 days of gestation. When congenital hypothyroidism was induced in rats, the post-natal disappearance of reactive fibers in the nodal regions was delayed. On the other hand, hypothyroidism induced in the adult rats did not change the number of the reactive nodal fibers with respect to the euthyroid hearts.

Animals↗
PubMed2001

Transient expression of P2X(1) receptor subunits of ATP-gated ion channels in the developing rat cochlea.

The expression pattern of the ATP-gated ion channel P2X(1) receptor subunit was studied in the developing rat cochlea by riboprobe in situ hybridisation and immunohistochemistry. Embryonic (E12, E14, E16 and E18) and postnatal (P0, P2, P4, P6, P10 and adult) rat cochleae were examined. Both mRNA and protein localisation techniques demonstrated comparable P2X(1) receptor expression from E16 until P6 but this expression was absent at later developmental stages. P2X(1) receptor mRNA expression was localised within the otic capsule and associated mesenchyme (from E16 to P6), spiral limbus (from P0 to P6) and within the spiral ligament adjacent to the insertion of Reissner's membrane (from P2 to P6). P2X(1) receptor protein had a similar distribution based upon immunoperoxidase localisation. P2X(1) receptor-like immunoreactivity was detected in the otic capsule and the surrounding mesenchyme (from E16 to P6), spiral limbus (from P0) and epithelial cells of Reissner's membrane (from P2 to P6). The spiral ganglion neurones showed the earliest P2X(1) receptor expression (from E16 to P6). This became associated with immunolabelling of their afferent neurite projections to the base of the developing inner and outer hair cells (observed from E18 and peaking at P2). Immunolabelling of the efferent nerve fibres of the intraganglionic spiral bundle (from E18 to P6) within the spiral ganglion was also observed. The results suggest that ATP-gated ion channels assembled from P2X(1) receptor subunits provide a signal transduction pathway for development of afferent and efferent innervation of the sensory hair cells and purinergic influence on cochlear morphogenesis.

Adenosine Triphosphate↗
PubMed1997

Promoters from Drosophila heat shock protein and cytomegalovirus drive transient expression of luciferase introduced by particle bombardment into embryos of the oyster Crassostrea gigas.

Using high velocity particle bombardment, we transferred a reporter gene into early stages of the oyster Crassostrea gigas and showed the expression of the introduced genes in these embryos at later stages of development. We tested two promoters: (1) the heat shock protein 70 promoter of Drosophila; (2) the cytomegalovirus early promoter, both linked to the luciferase reporter gene. The hsp 70-luc (pDrluc) construct allowed an expression level up to 55-fold higher than the control in a heat inducible fashion. The CMV-luc (pCMVL) construct constitutively gave a 4-fold higher expression than the control. This confirms the suitability of particle bombardment for transfecting genes into eggs, zygotes and trochopores of bivalves and demonstrates the functionality of two heterologous expression vectors in C. gigas.

Animals↗
PubMed2002

CIC, a member of a novel subfamily of the HMG-box superfamily, is transiently expressed in developing granule neurons.

We describe here the identification and characterization of a new gene, Cic, in both human and mouse genomes. These are orthologs of the Drosophila gene capicua, and represent a new subfamily of the HMG-box superfamily. Expression of the Cic gene is predominantly restricted to immature granule cells in the cerebellum, hippocampus and olfactory bulb in the CNS. This gene is therefore implicated in CNS development, in particular in granule cell development.

Amino Acid Sequence↗
PubMed2001

Transient expression of phospholipase D1 in developing rat hippocampus.

We investigated the distribution of phospholipase D1 (PLD1) protein in the developing rat hippocampus using an affinity-purified peptide antibody against PLD1. Immunoreactivity for PLD1 was first seen in some scattered cells in the hippocampus at embryonic day 18. At postnatal day 1 (P1), many PLD1 immunoreactive cells were observed in the CA1 and CA3 sectors, subiculum and the hilus of the dentate gyrus. During the first postnatal week, there was an abrupt increase of immunoreactive neurons in the hippocampus, and their number and intensity peaked at P7. During the second postnatal week, there was an abrupt decrease in the number of immunoreactive hippocampal neurons. By P14, no significant labeling was found in the hippocampus. These results corresponded well with those from Western blot analysis, suggesting that PLD1 may regulate the developmental processes of hippocampal neurons.

Animals↗
PubMed1998

Transient expression of calretinin in the trout habenulo-interpeduncular system during development.

Calcium-binding proteins control calcium homeostasis during neural development. The expression of one of these proteins, calretinin (CR), was monitored by immunohistochemistry in the developing habenulo-interpeduncular system of the rainbow trout, a conserved region of the brain along vertebrate phylogeny that undergoes a neurochemical reorganization in late development. No CR-immunoreactivity was observed in the habenulo-interpeduncular system during the embryonic development. CR-immunolabeling appeared in newly hatched fry and during the fry development the number of CR-immunostained elements increased progressively. During the juvenile stages (from 30 days post-hatching onwards) a gradual decrease in the number of CR-immunostained cells occurred, until its complete disappearance in adults. These variations in CR expression may represent the variable calcium-buffering needs during different developmental stages.

Age Factors↗