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Biomedical subjects

F E Bloom

Publications and source records attributed to F E Bloom.

At least 181 records · Page 10Linked to original sources

A single nucleotide difference in the gene for myelin proteolipid protein defines the jimpy mutation in mouse.

We have previously shown that, in the myelin-deficient jimpy mutant mouse, 74 nucleotides are absent from the mRNA for proteolipid protein (PLP) as a result of aberrant RNA processing. To define the exact site of the jimpy mutation, we have analyzed the PLP gene obtained from a jimpy mouse genomic library. We find that the nucleotide sequence that is absent from jimpy PLP mRNA is fully preserved in the jimpy PLP gene. The missing segment corresponds to a separate exon, equivalent to exon 5 of the human PLP gene. The nucleotide sequence at the 3' end of intron 4 in the jimpy PLP gene contains a single point mutation. A base change A----G in the 3' acceptor splice site has altered a position that is 100% conserved in all published splice acceptor sequences. We conclude that the primary genetic defect of the jimpy mouse is a single base change in the PLP gene disabling an invariant recognition sequence of RNA splicing.

Animals↗

Neural protein 1B236/myelin-associated glycoprotein (MAG) defines a subgroup of the immunoglobulin superfamily.

We have reviewed the structure and properties of the neural protein 1B236/MAG. This molecule consists largely of five Ig-like domains separated from its carboxyl terminal tail by a single membrane-spanning region. Two forms of the protein differ in the length and sequence of the carboxyl terminus: these are encoded by alternatively spliced mRNAs that are differentially expressed during postnatal neural development. The Ig-like domains of 1B236/MAG are unusual in having structural similarities to Ig V domains but with short Cys-Cys distances characteristic of C domains. Several other Ig-like molecules exhibit this structural feature, including the cell adhesion molecule N-CAM, which is most closely related in sequence to 1B236/MAG. We have proposed 1B236/MAG as the prototype for this subgroup of the Ig family and offer a model for this type of Ig domain structure. 1B236/MAG probably acts as a cell adhesion molecule to mediate interactions between cells in a fashion similar to that proposed for N-CAM. In particular, 1B236/MAG may be involved in interactions between myelinating oligodendrocytes or Schwann cells and axons or between adjacent layers of myelin membrane during the process of myelin compaction. It is most likely that the homophilic or heterophilic interactions of 1B236/MAG occur through binding to the Ig-like domains. The structure of 1B236/MAG is therefore quite consistent with its proposed function and may serve as the model for this class of cell-cell interaction molecules. One would predict, for example, that the neuron-glia cell adhesion molecule Ng-CAM, also known as NILE or L1 (Bock et al. 1985, Friedlander et al. 1985), which mediates interactions between neurons and glial cells, would have a very similar structure to those of N-CAM and 1B236/MAG. In addition, the carboxyl terminal tails of the 1B236/MAG proteins may also be involved in interactions with cytoskeletal components, during membrane vesicle transport through the glial cytoplasm during myelination or through neuronal axoplasm or cytoplasm. The availability of full-length cDNA clones of 1B236/MAG mRNAs with the ability to express these products in vitro will enable the structure and interactions of 1B236/MAG to be tested in detail.

Amino Acid Sequence↗

Electrophysiology of ethanol on central neurons.

With respect to the theme of this volume, the results of our recent studies on three neuronal model systems point to several relevant conclusions: ethanol may interact electrophysiologically with certain anesthetics such as urethane; ethanol can selectively enhance responses to certain neurotransmitters; resting membrane properties of individual neurons show a wide range of sensitivities to ethanol and are generally fairly insensitive; the synapse--independent of specific transmitters--seems most sensitive to ethanol. As regards the first point, it has long been known that ethanol and anesthetics have features in common, including the ability to alter the lipid components of biological membranes (see R. A. Harris et al., L. L. M. van Deenen et al., M. J. Hudspith et al., E. Rubin et al., and C. C. Cunningham & P. I. Spach in this volume), so interactions between the two are not unexpected. However, our electrophysiological findings suggest great caution and appropriate controls be used in in-vivo studies of anesthetized animals, as the interactions derived may actually reverse the usual effect of ethanol. The enhancement of responses to ACh and SS (second point) might be assumed to arise postsynaptically in the target cells recorded and are seen with low, intoxicating doses of ethanol. Whether this potentiation involves enhancement of specific agonist binding to the receptor or facilitation of the function of the ionic channel linked to the receptor remains to be determined. It is not hard to imagine that ethanol could perturb membrane properties near receptors, to alter their conformation and ligand binding, or perhaps even uncover hidden receptors. The relative insensitivity of the resting membrane properties (third point) may suggest that membrane channels responsible for these functions (e.g., 'leak' channels for Na+ and K+ ions) do not usually interact with the lipid components affected by ethanol, at least at low, 'intoxicating' ethanol concentrations. Finally, the reduction of synaptic potentials by ethanol may indicate a presynaptic locus of action, as the response to the transmitter for at least one of these synaptic potentials (GABA) was not altered. These data would seem to indicate that synaptic release of the transmitter is reduced by ethanol, at least in the hippocampal slice. The high sensitivity of this presynaptic element for ethanol could indicate that the machinery for synaptic release, such as conductances for calcium entry (see REF. 39) or the action of second messenger systems (e.g., those leading to synapsin phosphorylation) are particularly sensitive to ethanol.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗

Expression of small cytoplasmic transcripts of the rat identifier element in vivo and in cultured cells.

We examined the level of expression of small RNA transcripts hybridizing to a rodent repetitive DNA element, the identifier (ID) sequence, in a variety of cell types in vivo and in cultured mammalian cells. A 160-nucleotide (160n) cytoplasmic poly(A)+ RNA (BC1) appeared in late embryonic and early postnatal rat brain development, was enriched in the cerebral cortex, and appeared to be restricted to neural tissue and the anterior pituitary gland. A 110n RNA (BC2) was specifically enriched in brain, especially the postnatal cortex, but was detectable at low levels in peripheral tissues. A third, related 75n poly(A)- RNA (T3) was found in rat brain and at lower levels in peripheral tissues but was very abundant in the testes. The BC RNAs were found in a variety of rat cell lines, and their level of expression was dependent upon cell culture conditions. A rat ID probe detected BC-like RNAs in mouse brain but not liver and detected a 200n RNA in monkey brain but not liver at lower hybridization stringencies. These RNAs were expressed by mouse and primate cell lines. Thus, tissue-specific expression of small ID-sequence-related transcripts is conserved among mammals, but the tight regulation found in vivo is lost by cells in culture.

Animals↗

Clinical perspectives on neuropeptides.

Abnormalities in various neuropeptide systems have been detected in a number of neuropsychiatric disorders. Understanding the pathophysiological significance of these findings requires considering them in the context of the neural circuits involved. In both Huntington's disease and Alzheimer's disease, the reported alterations in multiple neuropeptide systems may help define the primary pathologic change, and may lead to new therapeutic strategies.

Alzheimer Disease↗

Developmentally regulated rat brain mRNAs: molecular and anatomical characterization.

In order to identify markers for developing neural cell populations and gain molecular insights into the processes of neural development and differentiation, we have selected cDNA clones of rat brain mRNAs that are expressed in brain at embryonic day 16 (E16) with at least 10-fold greater abundance than they are in adult brain. Eleven such clones were obtained from a cDNA library of E16 brain poly(A)+ RNA using a combination of differential and subtractive hybridization screens. The temporal and spatial patterns of expression of the mRNAs corresponding to these clones were characterized by Northern (RNA) blotting and by in situ hybridization. Although all the mRNAs were enriched in embryonic brain, different mRNAs demonstrated maximum abundance at different times in late embryogenesis. The mRNAs can be grouped into 3 classes on the basis of their patterns of spatial expression in the embryo: one cDNA clone from each class and its corresponding mRNAs have been characterized in more detail. Class C represents mRNAs that are highly enriched in the nervous system and may be expressed in newly differentiating neurons; the example chosen was shown by nucleotide sequence analysis to encode the brain alpha 1 isotype of tubulin. Class B mRNAs have a broader distribution in the developing embryo but are expressed predominantly in the ventricular germinal zones of the developing nervous system and may represent molecules involved with neurogenesis. A third class (Class A) includes mRNAs with a more homogeneous distribution within the embryo and developing nervous system, which may encode "housekeeping" molecules. These clones and their encoded products will provide markers for cell populations at particular stages of neural development.

Animals↗

Opioid enhancement of perforant path transmission: effect of an enkephalin analog on inhibition and facilitation in the dentate gyrus.

The enkephalin analogue [D-Ala2,D-Leu5]enkephalin, applied locally in vivo, enhanced the responsiveness of dentate granule cells of the hippocampus to perforant path stimulation. The facilitatory effect included a decrease in the inhibition and increase in the facilitation induced by sequential stimuli. The results indicate that opioids, acting at opioid receptors, can enhance responsiveness of the dentate gyrus to perforant path transmission, possibly via a disinhibitory mechanism.

Action Potentials↗

Systemic ethanol: selective enhancement of responses to acetylcholine and somatostatin in hippocampus.

In rat hippocampal pyramidal cells tested in situ by iontophoresis of several neurotransmitters, ethanol significantly enhanced excitatory responses to acetylcholine and inhibitory responses to somatostatin-14 but had no statistically significant effect on excitatory responses to glutamate or inhibitory responses to gamma-aminobutyric acid or, in preliminary tests, to norepinephrine or serotonin. The effects of ethanol on responses to acetylcholine and somatostatin-14 may provide insight into synaptic mechanisms underlying the behavioral consequences of ethanol intoxication.

Acetylcholine↗

P300 latency reflects the degree of cognitive decline in dementing illness.

An auditory discrimination paradigm was employed to elicit the P3 component of the event-related brain potential (ERP) from 39 demented patients (mean age = 71 years). Component latency was longer in patients who were diagnosed as having primary degenerative dementia and other cognitive impairment disorders compared to age-matched controls. Neurologist ratings of cognitive impairment were significantly correlated with P3 latency values, although no differences in mean latency were obtained between the various categories of dementia. ERP measurement techniques and the interpretation of P3 latency as in index of dementing illness are discussed.

Aged↗

Opiate antagonists do not alter neuronal responses to stimulation of opioid-containing pathways in rat hippocampus.

The opioid receptor antagonists, naloxone and beta-chlornaltrexamine, were used to determine whether activation of endogenous opioid peptide containing pathways produced pharmacologically reversible opioid actions. Extracellularly recorded responses of the hippocampal CA3 pyramidal cells were evoked by stimulation of the dynorphin-containing mossy fiber pathway. Neither naloxone nor beta-chlornaltrexamine pretreatment significantly changed the evoked response. However, both antagonists blocked the effect of applied dynorphin-A(1-17) on CA1 pyramidal cell evoked responses. Thus, our data demonstrate that if endogenous opioids are released from this pathway, the peptides cannot be responsible for the evoked response measured in hippocampal CA3 cellular field. With no direct evidence for endogenous opioid peptides acting through opioid receptors, the neurotransmitter role of dynorphins in rat hippocampus remains obscure.

Animals↗

P300 and alcohol consumption in normals and individuals at risk for alcoholism. A preliminary report.

Pairs of college student subjects (36 male, 36 female) were matched on age, sex, and personal drinking history. One pair member had a parent who met the DSM III criteria of alcoholism, while the other pair member had no close alcoholic relative. The P300 event-related brain potential (ERP) was obtained from each subject with auditory stimuli in an "oddball" paradigm. Target stimuli occurred randomly on 20% of the trials in a frequency discrimination task, a relatively easy intensity discrimination task, and a more difficult intensity discrimination task. Subjects indicated when the target items occurred by moving their index finger. No significant overall effects were obtained for family history for either P300 latency or amplitude. P300 latency increased and amplitude decreased with increases in the reported amount of alcohol consumption in all subjects only for the difficult intensity task but were statistically significant only for individuals with a negative family history for alcoholism.

Adolescent↗

Efferent projections of nucleus locus coeruleus: topographic organization of cells of origin demonstrated by three-dimensional reconstruction.

The present study examines the spatial distribution within rat locus coeruleus of neurons projecting to particular brain regions. In order to accurately recreate, in digital and pictorial formats, the spatial distribution of neurons for the entire nucleus locus coeruleus, three-dimensional reconstructions were created which specified the location of each individual Nissl-stained locus coeruleus cell in each of five nuclei. Dynamic computerized displays were visually analyzed and statistically compared. The nuclei from different brains were found to be strikingly similar in density and distribution of cells. In order to determine whether the cells of origin for particular locus coeruleus projections were clustered within the nucleus, reconstructions were created of the distributions of cells labeled by injections of a retrograde tracer, horseradish peroxidase, into particular terminal regions. Groups consisting of animals with injections into the same target areas were visually and statistically compared. The cells of origin of most efferent projections were found to be spatially organized within locus coeruleus. Specifically, projections to both the dorsal and ventral hippocampus originated solely from the dorsal segment of the nucleus, while spinal cord projections originated from ventral-posterior locus coeruleus. Cells of origin of cerebral and cerebellar cortical efferents, as well as hypothalamic efferents, exhibited less clustering, although reliable differences in distribution were observed. The most striking example of clustered cells of origin was exhibited by the large norepinephrine-containing cells constituting the anterior pole of locus coeruleus which were labeled only by hypothalamic injections. This analysis of spatial organization within locus coeruleus is unique in its utilization of a defined control group, experimental groups consisting of strictly defined replications, accurate three-dimensional reconstruction, and statistical comparisons. The demonstrated spatial heterogeneity of locus coeruleus neurons with respect to efferent projections can now be compared to the spatial distributions of other cellular characteristics such as soma morphology, colocalized transmitters and physiological properties. Presumably, such spatial segregation reflects the operation of functionally important organizing principles within the nucleus.

Animals↗

Somatostatin selectively enhances acetylcholine-induced excitations in rat hippocampus and cortex.

The neuronal effects of somatostatin-14 (SS-14) and its influence on responses to acetylcholine (AcCho) were studied in vivo in the rat parietal cortex and dorsal hippocampus, using single-unit recording and microiontophoresis. SS-14 inhibited spontaneous firing of nearly all cells tested, while AcCho facilitated their firing. In contrast to its direct slowing effect, sustained iontophoretic application of SS-14 enhanced AcCho-induced excitations in 78% of all cells tested. This AcCho-enhancing effect of SS-14 was dose dependent. SS-14 did not enhance the responsiveness to pulses of the excitatory amino acid glutamate. Neurons tonically driven by iontophoretic currents of AcCho responded to concurrent pulses of SS-14 with an increase in firing. Thus, iontophoretic application of SS-14 can produce qualitatively different effects on the spontaneous activity of its target cells depending on the simultaneous effects of other chemical messengers. These condition-dependent interactions may explain the diverse neuronal effects of SS-14 reported in the literature.

Acetylcholine↗

Jimpy mutant mouse: a 74-base deletion in the mRNA for myelin proteolipid protein and evidence for a primary defect in RNA splicing.

The mouse mutant jimpy carries an X chromosome-linked recessive gene defect that affects the formation of myelin in the central nervous system. To understand the molecular basis of the jimpy mutation, we have examined the expression of mRNAs encoding myelin proteolipid protein (PLP). PLP mRNAs were detectable in jimpy brain RNA at 21 days after birth but were severely reduced in abundance compared to wild-type littermates. Nucleotide sequence analysis of cDNA clones for PLP mRNA, isolated from a cDNA library of jimpy brain poly(A)+ RNA, revealed that the PLP mRNA expressed in jimpy contained a deletion of 74 nucleotides with respect to the wild-type sequence. This deletion causes a frameshift in the open reading frame resulting in an altered carboxyl terminus for jimpy PLP. Probes specific for the deleted sequence, however, hybridize with equal efficiency to genomic DNA from jimpy and wild-type littermates, suggesting that the defect in the jimpy PLP mRNA is generated by aberrant RNA processing rather than by deletion of genomic sequences. We conclude that a mutation in the gene for PLP that leads to an incorrectly spliced RNA transcript is the primary defect of this genetic disorder.

Amino Acid Sequence↗

Subcutaneous administration of behaviorally effective doses of arginine vasopressin change brain AVP content only in median eminence.

The accumulation of [8-arginine]vasopressin (AVP) in brain areas inside the blood-brain barrier (thalamus-hypothalamus, amygdala with overlying temporal cortex, hippocampus and cerebral cortex) and outside the blood-brain barrier (median eminence of the hypothalamus and area postrema) was measured after subcutaneous injection of the hormone. The plasma concentrations of AVP peaked at 5 min after subcutaneous injection and declined in a biphasic manner over the next 115 min. The concentration of AVP in brain tissue samples peaked at 20 min after the subcutaneous injection of AVP; the decline of AVP in the areas protected by the blood-brain barrier followed the time course seen for plasma. The concentration of AVP in the brain areas not so protected also peaked at 20 min but these declined at rates that differed from other brain areas and plasma. The concentration of AVP in the plasma and in most brain areas depended on the dose administered, while those in the median eminence and in the area postrema did not. Water deprivation for 24 and 48 h significantly elevated both the plasma AVP concentration and the concentration of AVP in the hypothalamus and in the amygdala-temporal cortex samples. The increases in AVP after water deprivation are limited to these two regions and are quantitatively much lower than after peripheral administration. Furthermore, when the brains of anesthetized rats were perfused free of blood, there were no changes in regional brain AVP content after subcutaneous treatment with 5,000 ng/kg of AVP, except for the median eminence. These data suggest that circulating AVP does not enter the parenchyma of brain areas protected by the blood-brain barrier in sufficient quantities to be detected by our assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗