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J I Morgan

Publications and source records attributed to J I Morgan.

At least 55 records · Page 3Linked to original sources
PubMed1995

A protein related to extracellular matrix proteins deleted in the mouse mutant reeler.

The autosomal recessive mouse mutation reeler leads to impaired motor coordination, tremors and ataxia. Neurons in affected mice fail to reach their correct locations in the developing brain, disrupting the organization of the cerebellar and cerebral cortices and other laminated regions. Here we use a previously characterized reeler allele (rl(tg)) to close a gene, reelin, deleted in two reeler alleles. Normal but not mutant mice express reelin in embryonic and postnatal neurons during periods of neuronal migration. The encoded protein resembles extracellular matrix proteins involved in cell adhesion. The reeler phenotype thus seems to reflect a failure of early events associated with brain lamination which are normally controlled by reelin.

Alleles↗
PubMed1995

Fos: an immediate-early transcription factor in neurons.

In the past several years a great deal of evidence has accumulated linking neuronal activation events to the regulation of gene expression. We have pursued an analysis of c-fos regulation in the nervous system to elucidate the molecular mechanisms involved in stimulus-transcription coupling. The c-fos gene can be viewed as an archetype of the set of cellular immediate-early genes encoding transcription factors. These genes are believed to function in coupling short-term signals elicited by extracellular to long-term changes in cellular phenotype by orchestrating alterations in target gene expression. Several animal seizure models have been used to demonstrate the activation of gene expression in specific populations of neurons. Using a transgenic mouse approach, based on a foslacZ fusion gene, we now demonstrate an association between c-fos expression and cell death in the nervous system. A delayed and protracted induction was observed following surgical lesion and in response to neurotoxin exposure. This system allows us to determine, for the first time, the DNA regulatory sequences that are responsible for the induction of gene expression in neurons in vivo. Furthermore, foslacZ transgenic mice provide a unique resource for identifying cell populations that respond to specific stimuli or that are susceptible to particular toxins.

Animals↗
PubMed1995

The cellular prion protein (PrP) selectively binds to Bcl-2 in the yeast two-hybrid system.

Bcl-2 can rescue neurons from death and might, therefore, exert its action by associating with neuron-specific proteins. Using LexA-Bcl-2 as bait, we find that the cellular prion protein (PrP) interacts with Bcl-2, but not Bax, in the yeast two-hybrid system. Since the PrP gene has been implicated in neurodegenerative disorders, this preliminary observation suggests a potential pathogenic mechanism for these conditions.

Apoptosis↗
PubMed1995

Regulation of c-fos expression in transgenic mice requires multiple interdependent transcription control elements.

Transcription control regions of eukaryotic genes contain multiple sequence elements proposed to function independently to regulate transcription. We developed transgenic mice carrying fos-lacZ fusion genes with clustered point mutations in each of several distinct regulatory sequences: the sis-inducible element, the serum response element, the fos AP-1 site, and the calcium/cAMP response element. Analysis of Fos-lacZ expression in the CNS and in cultured cells demonstrated that all of the regulatory elements tested were required in concert for tissue- and stimulus-specific regulation of the c-fos promoter. This implies that the regulation of c-fos expression requires the concerted action of multiple control elements that direct the assembly of an interdependent transcription complex.

Animals↗
PubMed1995

The olfactory system as a model for the analysis of the contribution of gene expression to programmed cell death.

The process of programmed cell death is frequently attenuated by inhibitors of protein and RNA synthesis. This implies that gene expression is necessary for the active elimination of some cell types. Genes such as bcl-2 and bax have been implicated in the direct control of cell death, while cellular immediate-early genes (cIEGs), such as c-fos and c-jun have been repeatedly associated with neuronal degeneration. We are using the olfactory neuroepithelium as a model system to investigate the role that expression of such genes might play in cell death. The advantages of this system is that even in the adult, there is spontaneous degeneration of olfactory receptor neurons followed by their replacement by the division and differentiation of precursors. Furthermore, the receptor neurons can be induced to die synchronously by removal of the olfactory bulb or intranasal administration of toxic agents. We have generated fos-lacZ and jun-lacZ transgenic mice that can be used to assess expression of c-fos and c-jun following these various manipulations. In addition, a line of transgenic mice has been derived that express Bcl-2 under the control of the olfactory receptor protein promoter. These mice have high levels of Bcl-2 selectively in receptor neurons of the primary neuro-epithelium and vomeronasal organ. Since in some circumstances, Bcl-2 can protect against programmed cell death these mice are being assessed for neuronal turnover under basal conditions and following olfactory bulbectomy.

Animals↗
PubMed1995

The maf proto-oncogene stimulates transcription from multiple sites in a promoter that directs Purkinje neuron-specific gene expression.

L7 is expressed in all adult cerebellar Purkinje cells, although during development it appears in a stereotyped spatial and temporal pattern that is manifested as parasagittal domains of neurons. Mutations of the L7 promoter in transgenic mice have established that these domains represent functional compartments of Purkinje neurons. Therefore, it is hoped that by defining the transcriptional control of the L7 gene insights into the mechanisms that control functional fate and organization in the nervous system can be gained. Fragments of the L7 promoter were introduced into a selectable reporter gene in Saccharomyces cerevisiae, and these strains were used to select for cerebellar cDNAs encoding proteins that can bind to, and activate transcription from, these elements. This assay identified the c-Maf proto-oncogene as activating transcription from two sites in the L7 promoter. We did a functional domain analysis of vertebrate c-Maf based upon transcriptional activation in S. cerevisiae and showed the requirement for a transactivation domain, leucine zipper, and DNA-binding region in c-Maf. The c-Maf interaction site was mapped to the sequence G/TGG/CNG/TNCT CAGNN in the L7 promoter, which represents an atypical 12-O-tetradecanoate-13-acetate-responsive element-type Maf-responsive element. However, neither Fos nor Jun, either alone or in combination with each other or c-Maf, altered transcription from this element. In contrast, a Maf-related protein, Nrl, completely mimicked c-Maf actions. These data suggest that Maf may interact with additional basic-zipper proteins that determine a subtype of Maf-responsive element binding.

Animals↗
PubMed1995

Kainic acid-induced neuronal death is associated with DNA damage and a unique immediate-early gene response in c-fos-lacZ transgenic rats.

Previously, we established that persistent upregulation of c-fos expression preceded kainic acid (KA)-induced neuronal death in mice. To discriminate between events that are products of the seizures elicited by KA and those that are specifically associated with its neurotoxic actions, we have examined the expression of cellular immediate-early genes (cIEGs) following KA or pentylenetetrazol (PTZ) treatment in c-fos-lacZ transgenic rats. While both chemoconvulsants elicit seizures, only KA causes selective neuronal death. Following treatment of transgenic rats with KA there was a protracted expression of Fos-lacZ that lasted for 2-3 d. In contrast, PTZ elicited a transient increase in the transgene product that lasted about 6 hr. Normally, Fos and Fos-lacZ were detected only in neuronal nuclei. However, 6 hr following kainic acid (but not PTZ) administration, beta-galactosidase activity appeared in the cytoplasm of neurons within vulnerable regions (as determined by the terminal transferase biotinylated-UTP nick end labeling (TUNEL) procedure). Like c-fos, transcripts for other cIEGs were elevated for longer periods in the KA-treated rat hippocampus. In addition, fra-1 and fra-2 were only induced in the KA-treated rat. These changes in mRNA levels were paralleled by a sustained increase in AP-1 DNA binding activity. Thus, quantitative and qualitative changes in AP-1 DNA binding complexes accompany neurotoxic cell death that are not observed following seizures.

Animals↗
PubMed1994

Isolation of an allele of reeler by insertional mutagenesis.

Reeler (rl) is an autosomal recessive mutation that affects migration of postmitotic neurons in the mouse central nervous system. The reeler (rl/rl) mouse displays a disruption of laminar structures in both the cerebellum and the forebrain and it exhibits tremors, dystonia, and ataxia. The molecular basis of the reeler phenotype is unknown because the gene involved has not yet been identified. We report here the isolation and characterization of an allele of rl, reelertransgene (rltg). This allele was generated by the fortuitous insertion of a transgene, supfos (sf), into the mouse rl locus. Crosses between rl/+ and rltg/+ mice yielded offspring that exhibited the reeler phenotype, indicating that rl and rltg are allelic. We cloned the genomic sequences flanking the transgene insertion site from the rltg/rltg mouse genome. Chromosomal mapping studies revealed that the 5' flanking cellular sequence maps to a locus, D5Gmr1, that lies in a region of mouse chromosome 5 that also contains the rl locus. Southern blot analysis using a probe derived from the D5Gmr1 locus revealed no gross structural rearrangement in the rl locus. Thus, unlike the two rl alleles described previously, rltg provides a molecular probe that can now be used to identify and isolate the rl gene.

Alleles↗
PubMed1994

A third synaptotagmin gene, Syt3, in the mouse.

The synaptotagmins are integral membrane proteins of synaptic vesicles thought to serve as Ca2+ sensors in the process of vesicular trafficking and exocytosis. Results from antibody microinjection and gene-disruption experiments have led to a controversy over whether synaptotagmins are essential for neurotransmission. However, the studies casting doubt on the role of synaptotagmins have assumed that no further isoforms of these molecules exist. Here we report the isolation of a third member of the synaptotagmin family (Syt3) from mouse brain. Although retaining the characteristic five-domain structure of the other synaptotagmins, SYT3 is considerably more divergent at the level of amino acid sequence. In the most highly conserved C2 domain, the mammalian synaptotagmins, SYT1 and SYT2, share 88% sequence identity, whereas SYT3 has only approximately 45% identity to either. Overall, SYT3 has the greatest sequence identity with rat SYT2 and marine ray p65A (both 37%), although homology to all of the known synaptotagmins is > 30%. However, SYT3 is most like p65C when comparing domain structure. Syt3 is expressed in many regions of the nervous system but is undetectable in extraneural tissues. The three murine synaptotagmins have differential expression patterns in the brain. Furthermore, in PC12 cells, Syt3 is coexpressed with Syt1 and is more abundant than the latter. This result suggests that individual neurons may have specific combinations of synaptotagmins that could provide for diversity in vesicular release.

Amino Acid Sequence↗
PubMed1994

Genomic structure and mapping of precerebellin and a precerebellin-related gene.

The cerebellum-specific hexadecapeptide, cerebellin, is derived from a larger precursor, precerebellin, that has sequence homology to the complement component C1qB. We report the cloning of the murine homolog of precerebellin, Cbln1, and a closely related gene, Cbln2. Amino acid comparison of Cbln1 with Cbln2 revealed that Cbln2 is 88% identical to the carboxy terminal region of Cbln1. That these are independent genes was confirmed by Southern analysis and genome mapping. Cbln1 was positioned to the central region of mouse chromosome 8, 2.3 cM distal of JunB and 6.0 cM proximal of Mt1, while Cbln2 mapped to the distal end of mouse chromosome 18, 1.7 cM telomeric of Mbp.

Amino Acid Sequence↗
PubMed1994

Targeted disruption of NMDA receptor 1 gene abolishes NMDA response and results in neonatal death.

In vitro studies have suggested that the NMDA receptor consists of an essential subunit, NR1, and various modulatory NR2 subunits. To test this hypothesis directly in vivo, we generated mice carrying a disrupted NR1 allele. NMDA-inducible increases in intracellular calcium and membrane currents were abolished in neurons from homozygous null mutants (NR1-/-). Thus, NR1 has a unique role, which cannot be substituted by any other subunit, in determining the activity of the endogenous NMDA receptor. A concomitant reduction in levels of NR2B but not NR2A occurred in NR1-/- mice, demonstrating that there is an interdependence of subunit expression. NR1-/- mice died 8-15 hr after birth, indicating a vital neonatal function for the NMDA receptor. Although the NMDA receptor has been implicated in several aspects of neurodevelopment, overall neuroanatomy of NR1-/- mice appeared normal. Pathological evidence suggested that respiratory failure was the ultimate cause of death.

Animals↗
PubMed1993

Continuous c-fos expression precedes programmed cell death in vivo.

The development of a multicellular organism involves a delicate balance among the processes of proliferation, differentiation and death. Naturally occurring cell death aids tissue remodelling, eliminates supernumerary cell populations and provides structural elements such as hair and skin. In the nervous system, selective cell death contributes to the formation and organization of the spinal cord and sympathetic ganglia, retina and corpus callosum. But cell death also occurs in several neuropathological conditions, such as amyelotrophic lateral sclerosis and Alzheimer's disease. Therefore an elucidation of the mechanisms responsible for cell death is critical for an appreciation of both normal development and neuropathological disorders. Using a fos-lacZ transgenic mouse, we provide evidence showing that the continuous expression of Fos, beginning hours or days before the morphological demise of the cell, appears to be a hallmark of terminal differentiation and a harbinger of death.

3T3 Cells↗
PubMed1993

Structure and regulation of the gene encoding the neuron-specific protein PEP-19.

PEP-19 is a 61 amino acid polypeptide that is localized to neurons. PEP-19 is translated from a 0.6 kb poly(A)+ RNA; however, the gene from which it is transcribed spans more than 30 kbp and comprises three exons and two large introns. Exon 1 contains the 5'-untranslated region of PEP-19 and the first three amino acids of the coding sequence; exon 2 codes for the next 17 amino acids while the majority of the PEP-19 mRNA, comprising the remaining 42 amino acids and the 3'-untranslated region, is in exon 3. Gel retardation analysis identifies a region of DNA in the putative promoter of PEP-19 that binds preferentially to nuclear extracts from cerebellum. However, constructs containing 1.35 kbp of 5'-upstream genomic DNA of PEP-19 fused to lacZ do not express in transgenic mice, suggesting that intragenic DNA may be essential for the regulation of PEP-19.

Amino Acid Sequence↗
PubMed1993

Control of segment-like patterns of gene expression in the mouse cerebellum.

A Purkinje cell-specific transgene, L7-lacZ, is expressed in a series of parasagitally oriented stripes in the mouse cerebellum. This banding pattern can be perturbed by promoter mutation, showing that a combination of positive and negative control elements contributes to the temporal and spatial map of L7 gene expression. In addition to the parasagittal stripes, certain mutations reveal Purkinje cells organized into compartments oriented in the transverse plane of the cerebellum. Transcription factors of the POU or homeobox families appear to be involved in controlling L7 expression in the transverse orientation. Strikingly, some of the domains of gene expression revealed by the mutations appear to correspond to functional compartments of Purkinje cells, thereby suggesting an underlying genetic principle used to orchestrate functional organization in the nervous system.

Animals↗