Search PubMed⌕ Search

Biomedical subjects

M Segal

Publications and source records attributed to M Segal.

At least 109 records · Page 6Linked to original sources
PubMed1996

Development of neuronal precursor cells and functional postmitotic neurons from embryonic stem cells in vitro.

To understand the mechanism of the sequential restriction of multipotency of stem cells during development, we have established culture conditions that allow the differentiation of neuroepithelial precursor cells from embryonic stem (ES) cells. A highly enriched population of neuroepithelial precursor cells derived from ES cells proliferates in the presence of basic fibroblast growth factor (bFGF). These cells differentiate into both neurons and glia following withdrawal of bFGF. By further differentiating the cells in serum-containing medium, the neurons express a wide variety of neuron-specific genes and generate both excitatory and inhibitory synaptic connections. The expression pattern of position-specific neural markers suggests the presence of a variety of central nervous system (CNS) neuronal cell types. These findings indicate that neuronal precursor cells can be isolated from ES cells and that these cells can efficiently differentiate into functional post-mitotic neurons of diverse CNS structures.

Animals↗
PubMed1996

A recombination outside the BB deletion refines the location of the X linked retinitis pigmentosa locus RP3.

Genetic loci for X-linked retinitis pigmentosa (XLRP) have been mapped between Xp11.22 and Xp22.13 (RP2, RP3, RP6, and RP15). The RP3 gene, which is responsible for the predominant form of XLRP in most Caucasian populations, has been localized to Xp21.1 by linkage analysis and the map positions of chromosomal deletions associated with the disease. Previous linkage studies have suggested that RP3 is flanked by the markers DXS1110 (distal) and OTC (proximal). Patient BB was thought to have RP because of a lesion at the RP3 locus, in addition to chronic granulomatous disease, Duchenne muscular dystrophy (DMD), mild mental retardation, and the McLeod phenotype. This patient carried a deletion extending approximately 3 Mb from DMD in Xp21.3 to Xp21.1, with the proximal breakpoint located approximately 40 kb centromeric to DXS1110. The RP3 gene, therefore, is believed to reside between DXS1110 and the proximal breakpoint of the BB deletion. In order to refine the location of RP3 and to ascertain patients with RP3, we have been analyzing several XLRP families for linkage to Xp markers. Linkage analysis in an American family of 27 individuals demonstrates segregation of XLRP with markers in Xp21.1, consistent with the RP3 subtype. One affected mate shows a recombination event proximal to DXS1110. Additional markers within the DXS1110-OTC interval show that the crossover is between two novel polymorphic markers, DXS8349 and M6, both of which are present in BB DNA and lie centromeric to the proximal breakpoint. This recombination places the XLRP mutation in this family outside the BB deletion and redefines the location of RP3.

Adult↗
PubMed1996

Dynamic analysis of the three-phasic vascular response to histamine in coronary circulation.

In this work we tried to perform a dynamic analysis of coronary vascular response to histamine, using a simple mathematical analysis as a first step in the better understanding of complex histamine effects on coronary blood vessels of the isolated guinea pig heart. So, we defined more parameters, such as: I and delta I to quantify the intensity of coronary perfusion pressure (CPP) change, T and dt to quantify the time CPP change occurs and D to characterize the intensity and duration of CPP change, i.e. to quantify the dynamics of this change.

Animals↗
PubMed1995

Differential activation of yeast adenylyl cyclase by Ras1 and Ras2 depends on the conserved N terminus.

Although both Ras1 and Ras2 activate adenylyl cyclase in yeast, a number of differences can be observed regarding their function in the cAMP pathway. To explore the relative contribution of conserved and variable domains in determining these differences, chimeric RAS1-RAS2 or RAS2-RAS1 genes were constructed by swapping the sequences encoding the variable C-terminal domains. These constructs were expressed in a cdc25ts ras1 ras2 strain. Biochemical data show that the difference in efficacy of adenylyl cyclase activation between the two Ras proteins resides in the highly conserved N-terminal domain. This finding is supported by the observation that Ras2 delta, in which the C-terminal domain of Ras2 has been deleted, is a more potent activator of the yeast adenylyl cyclase than Ras1 delta, in which the C-terminal domain of Ras1 has been deleted. These observations suggest that amino acid residues other than the highly conserved residues of the effector domain within the N terminus may determine the efficiency of functional interaction with adenylyl cyclase. Similar levels of intracellular cAMP were found in Ras1, Ras1-Ras2, Ras1 delta, Ras2, and Ras2-Ras1 strains throughout the growth curve. This was found to result from the higher expression of Ras1 and Ras1-Ras2, which compensate for their lower efficacy in activating adenylyl cyclase. These results suggest that the difference between the Ras1 and the Ras2 phenotype is not due to their different efficacy in activating the cAMP pathway and that the divergent C-terminal domains are responsible for these differences, through interaction with other regulatory elements.

Adenylyl Cyclases↗
PubMed1995

Imaging of calcium variations in living dendritic spines of cultured rat hippocampal neurons.

1. Cultured rat hippocampal neurons were loaded with the Ca2+ indicator fura-2 through micropipettes and visualized with an inverted microscope equipped with a high power objective and a cooled CCD camera. The responses of dendritic spines and their parent dendrites to stimuli which evoke a rise of [Ca2+]i were monitored. 2. NMDA caused a rapid and transient rise in [Ca2+]i, which was more evident in the spine than in the parent dendrite. The recovery in both compartments had the same time course, and was dependent on normal [Na+]o. 3. Application of alpha-latrotoxin, which causes release of neurotransmitters from terminals, produced a rise of [Ca2+]i in the dendritic spines, more than in their parent dendrites. Prolonged exposure to the drug eliminated the spine/dendrite disparity. 4. The presence of voltage-gated calcium channels in dendritic spines is indicated by the enhanced calcium rise in spines rather than dendrites of cells depolarized by either intracellular current injection or by raising [K+]o. This rise was attenuated by nifedipine or verapamil, both L-type channel blockers. 5. It is suggested that the dendritic spine constitutes an independent calcium compartment that is closely linked to the parent dendrite.

Animals↗
PubMed1995

Morphological alterations in dendritic spines of rat hippocampal neurons exposed to N-methyl-D-aspartate.

Individual cultured rat hippocampal neurons grown on glass coverslips were loaded with lucifer yellow (LY) and segments of their dendrites, containing dendritic spines, were exposed to the glutamate agonist N-methyl-D-aspartate (NMDA). Repeated pulse application of NMDA caused a reduction of spine length by 20%, within 1-2 h of exposure to the drug. No such changes were seen in spines exposed to NMDA in presence of the antagonist 2-aminophosphonovalerate or in spines on sister dendrites which were not exposed to NMDA. It is suggested that dendritic spines shrink following exposure to excessive synaptic stimulation.

Animals↗
PubMed1995

Two distinct regions of Ras participate in functional interaction with GDP-GTP exchangers.

We have previously implemented a combined genetic/biochemical approach, for analysis of insertion-deletion mutants, to identify sites of Harvey-Ras participating in the interaction with guanine nucleotide exchangers, using the yeast Cdc25 as a model exchanger. We showed that positions 101-106 may be required for catalyzed exchange. We here present a further improved strategy to define more precisely the residues on Ras participating in this interaction. Non-conservative replacements at positions 103 or 105 abolished response to Cdc25 while substitutions at positions 102 or 104 were partially affected. The same substitutions had no effect on coupling to adenylyl cyclase. Since the strategy enables us to assess Ras functional interaction with both the exchanger and effector simultaneously, we have also examined the effect of substitutions in the distal part of the switch II region (amino acids 69-78). In contrast to other reports, substitutions at positions 69 or 73 prevented Cdc25 response while mutations at position 74 did not prevent this interaction. However, all these substitutions partly affected cyclase activation. These findings establish the crucial role of the 102-105 region in the catalyzed exchange reaction and suggest that the 69-74 area would be required for the functional interaction with both exchangers and effector molecules.

Adenylyl Cyclases↗
PubMed1995

[Testicular metastasis from transitional cell carcinoma of the bladder].

We present a 54-year-old man with a testicular metastasis originating from a transitional cell carcinoma of the urinary bladder. Initially, radical cystectomy and ileal-conduit urinary diversion were performed, followed 9 months later by urethrectomy for local recurrence. A year later he presented with a tumorous right testis proved to harbor transitional cell carcinoma.

Carcinoma, Transitional Cell↗
PubMed1995

Myxoid leiomyosarcoma of the uterus with unusual malignant histologic pattern--a case report.

A case report of myxoid leiomyosarcoma of the uterus is presented as a rare variant of uterine sarcoma. This case, contrary to most of the 11 cases previously described in the literature, is characterized by highly atypical pleomorphic cells and numerous mitoses. Furthermore, myxoid material is also found in the cytoplasm of pleomorphic tumor cells, a feature, that to our knowledge, has not been previously described. All these tumors, no matter what the microscopic appearance is, exhibit a highly malignant behavior. The best therapy for this tumor is unknown, since there are too few cases from which to draw conclusions.

Aged↗
PubMed1995

Imaging of calcium inside dendrites and spines: goals, issues and perspectives.

The growing interest in dendritic spines in recent years originates from both the realization that the spine is likely to be the site where long-term plastic changes in synaptic properties take place, and that imaging methods are available which allow, for the first time, the study of these changes in the living dendritic spine. This report briefly summarizes methodological and biological issues associated with the study of dendritic spines in living tissue. The combined use of electrical and imaging methods for the study of dendritic spines certainly will contribute to a better understanding of synaptic integration and plasticity.

Animals↗
PubMed1995

Dendritic spines for neuroprotection: a hypothesis.

Ever since their first description in neurons, dendritic spines could be visualized only in fixed tissue, using high-power light and electron microscopy. Recent studies have been able to measure the free intracellular Ca2+ concentration ([Ca2+]i) in dendritic spines of live neurons, and the results suggest that the spine is an independent cellular Ca2+ compartment. Other recent observations have indicated that the density of spines on dendrites changes in a dynamic fashion depending on ongoing neuronal activity. Together, these findings have led to the proposal that the dendritic spine is not only a storage device for long-term memory but perhaps a means for isolating the cell from the harmful consequences of synaptically evoked surges in [Ca2+]i. In other words, the dendritic spine is a neuroprotectant. This hypothesis has specific testable implications, including relating cell activity to spine density.

Animals↗
PubMed1995

0.0625% bupivacaine with 0.0002% fentanyl via patient-controlled epidural analgesia for pain of labor and delivery.

OBJECTIVE: To compare the utility of 0.0625% bupivacaine with fentanyl administered via patient-controlled epidural analgesia (PCEA) to a traditional continuous epidural infusion for pain of labor and delivery. DESIGN AND SUBJECTS: Forty-one women in established labor were randomized to receive either (a) 0.0625% bupivacaine with 2 micrograms/ml fentanyl via PCEA (demand dose = 3 ml, lockout interval = 6 min, background infusion = 6 ml/h, no 1 or 4 h limits) or (b) 0.125% bupivacaine with 2 micrograms/ml fentanyl via continuous epidural infusion (CEI) at 12 ml/h. Supplemental 0.25% bupivacaine (3 ml every 5 min, p.r.n., x 3) was administered for treatment of breakthrough pain upon patient request. The study protocol was double-blind and placebo-controlled. OUTCOME MEASURES: Visual analogue pain scores, motor strength, pinprick level of sensory analgesia and bupivacaine use were assessed by an anesthesiologist unaware of the individual patient's randomization to a particular study group. RESULTS: The cephalad extent of pinprick sensory analgesia was significantly lower during both the first (p < 0.03) and second (p < 0.03) stages of labor in patients receiving PCEA. However, visual analogue pain scores, intensity of motor blockade, and need for physician-administered supplemental bupivacaine were comparable in both groups. Patients receiving PCEA used 40% less bupivacaine per hour while achieving analgesia comparable to patients receiving CEI. CONCLUSIONS: The results of this study show that 0.0625% bupivacaine with 2 micrograms/ml of fentanyl is an effective analgesic combination when used via PCEA.

Adult↗
PubMed1995

Fast imaging of [Ca]i reveals presence of voltage-gated calcium channels in dendritic spines of cultured hippocampal neurons.

1. Cultured hippocampal neurons were recorded with a patch pipette containing 100 microM of the calcium indicator Fluo-3, and one of their dendrites, carrying dendritic spines, was visualized with a x100, 1.3-numerical aperture oil objective. Calcium spikes evoked by depolarizing the somata and changes in free dendrite and spine calcium concentrations ([Ca]d and [Ca]s, respectively) were monitored with a cooled charge-coupled device (CCD) camera, acquiring images at a rate of 17-20 ms per frame. In the majority of spine-dendrite pairs, [Ca]s rose faster and to a higher level than the adjacent [Ca]d. Likewise, topical application of glutamate evoked a faster and larger change in [Ca]s than in [Ca]d. The rise of intracellular calcium concentration in response to a depolarizing current pulse, but not in response to glutamate, was reduced in the presence of the calcium antagonist verapamil in both dendrites and spines. It is suggested that dendritic spines possess voltage-gated calcium channels.

Aniline Compounds↗
PubMed1995

[Hereditary hemorrhagic telangiectasia presenting with hematuria and severe anemia].

A patient with hereditary hemorrhagic telangiectasia was admitted with hematuria and severe anemia after mild recurrent episodes of epistaxis. Telangiectasias were found in the skin and buccal and nasal mucosa. No defect in the coagulation mechanism was found; thrombocyte count and function were normal. On cystoscopy, tortuous engorged vessels, some actively bleeding, were seen in the trigonal mucosa. Biopsy showed enlarged vessels in the lamina propria. Electrocoagulation of the bleeding vessels stopped hematuria, but 6 months later it recurred. This time Nd-YAG laser was used to stop the bleeding after electrocoagulation was ineffective.

Adult↗
PubMed1995

Morphological analysis of dendritic spine development in primary cultures of hippocampal neurons.

We monitored developmental alterations in the morphology of dendritic spines in primary cultures of hippocampal neurons using confocal laser scanning microscopy (CLSM) and the fluorescent marker Dil. Dissociated rat hippocampal neurons were plated on polylysine-coated glass cover slips and grown in culture for 1-4 weeks. Fixed cultures were stained with Dil and visualized with the CLSM. Spine density, spine length, and diameters of spine heads and necks were measured. Some cultures were immunostained for synaptophysin and others prepared for EM analysis. In the 1-3 week cultures, 92-95% of the neurons contained spiny dendrites. Two subpopulations of spine morphologies were distinguished. At 1 week in culture, "headless" spines constituted 50% of the spine population and were equal in length to the spines with heads. At 2, 3, and 4 weeks in culture headless spines constituted a progressively smaller fraction of the population and were, on average, shorter than spines with heads. Spines with heads had narrower necks than headless spines. At 3 weeks in culture, spines were associated with synaptophysin-immunoreactive labeling, resembling synaptic terminals. At 4 weeks in culture, only 70% of the Dil-filled cells had spiny dendrites, and the density of spines decreased. Ultrastructurally, the majority of dendritic spine-like structures at 1 week resembled long filopodia without synaptic contacts. The majority of axospinous synapses were on short "stubby" spines. At 3 weeks in culture, the spines were characteristic of those seen in vivo. They contained no microtubules or polyribosomes, were filled with a characteristic, filamentous material, and formed asymmetric synapses. These studies provide the basis for further analysis of the rules governing the formation, development, and plasticity of dendritic spines under controlled, in vitro conditions.

Animals↗
PubMed1994

Cdc25 is not the signal receiver for glucose induced cAMP response in S. cerevisiae.

The Ras/cAMP pathway in the yeast S. cerevisiae couples the cell cycle of this unicellular organism to the availability of nutrients. Glucose derepressed S. cerevisiae cells respond to glucose addition by an intracellular rise in cAMP. In the prevailing model, yeast Ras plays a similar role to that of heterotrimeric G-proteins coupled to cell surface receptors. A crucial element of this model is that the exchanger, Cdc25 is activated by glucose. Such activation would result in a glucose-dependent rise in GTP-bound Ras concentration. We here show, in contrast to this view, that Cdc25 cannot be the receiver of the glucose signal. We suggest that the Ras-GTP/cyclase complex is the molecular element directly receiving the signal while Cdc25-dependent exchange constitutes a prerequisite for complex formation.

Adenylyl Cyclases↗
PubMed1994

Acetylcholine mediates the effects of fenfluramine on dentate granule cell excitability in the rat.

Reactivity of the hippocampal system to stimulation of its main afferent, the perforant path, was studied in the intact, anesthetized rat. Parentral administration of fenfluramine caused a marked elevation of population spike response to perforant path stimulation. An injection of atropine before, but not after fenfluramine, blocked the potentiating effect of fenfluramine. The atropine blockade was dose-dependent and not mimicked by the peripheral muscarinic receptor antagonist methyl atropine. This effect of fenfluramine was also prevented by an injection of the 5-HT receptor antagonist spiperone. The effect of fenfluramine was mimicked by the anticholinesterase physostigmine, which was not affected by spiperone pretreatment. It is proposed that release of 5-HT (5-hydroxytryptamine) by fenfluramine potentiates reactivity to afferent stimulation by interacting with cholinergic terminals in the hippocampus.

Acetylcholine↗