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Biomedical subjects

A J Patel

Publications and source records attributed to A J Patel.

At least 37 records · Page 2Linked to original sources
PubMed1997

Presenilins and early-onset familial Alzheimer's disease.

Thirty-seven missense mutations and a splice-site mutation in the presenilin gene PS1 on chromosome 14 and two missense mutations PS2 on chromosome 1 co-segregate with early-onset familial Alzheimer's disease (AD). The presenilins belong to a family of conserved integral membrane proteins which include Caenorhabditis elegans SPE4 and SEL12 and the rat apoptosis-linked gene, ALG3. This review summarizes the genetics of presenilins in AD and indicators of putative function based on cellular localization and the functions of non-human homologues. Findings to date suggest an important role of presenilins in beta-amyloid (A beta) production: in vitro and in vivo studies have shown that presenilin mutations are associated with relatively increased production of the longer, and highly fibrillogenic A beta 42(43) peptide, and a marked elevation in the number of A beta 42-immunoreactive plaques in the brains of individuals with familial AD who carry PS1 and PS2 mutations. There is growing evidence that the deposition of A beta 42(43) could in some cases be an early and key event in the AD pathogenic cascade. The genetic and molecular biological data discussed in this review describe mechanisms by which presenilin mutations could lead to the development of AD. Also, mutant presenilins may be more proapoptotic. It is argued that the understanding of the processes by which presenilin mutations lead to the development of AD will help in devising a coherent framework for therapeutic strategies.

Adult↗
PubMed1997

Expression of beta-amyloid precursor protein immunoreactivity in the retina of the rat during normal development and after neonatal optic tract lesion.

Immunoreactivity to beta-amyloid precursor protein (APP) was present in the inner plexiform, ganglion cell and optic fibre layers, as well as in blood vessels, at birth in normally developing rat retinas. In the inner plexiform layer immunoreactivity disappeared by postnatal day (P) 14. A small population of ganglion cells was immunoreactive at birth, but none were visible at P7. From P14 onwards, however, there was weak immunoreactivity in ganglion cells again, and strong staining in Müller glia. Retinas affected by neonatal optic tract lesions contained more immunoreactive ganglion cells at P4 than did controls, but by P14 there was a severe loss of ganglion cells. These observations are consistent with APP being involved in retinal differentiation, including maturation of glia and neurones, synaptogenesis and possibly neuronal survival.

Aging↗
PubMed1997

Cell-specific expression of beta-amyloid precursor protein isoform mRNAs and proteins in neurons and astrocytes.

The abnormal accumulation of beta-amyloid (A beta) in senile plaques appears to be a central pathological process in Alzheimer's disease. A beta is formed by proteolysis of beta-amyloid precursor protein (APP) with several isoforms generated by alternative splicing of exons 7, 8 and 15. A semi-quantitative reverse transcription (RT)-polymerase chain reaction (PCR) analysis showed that APP695 mRNA lacking exon 7 and 8 was most abundant in primary cultures of rat neurons, while APP770 and APP751 representing, respectively, the full length and exon 8 lacking isoforms predominated in cultured astroglial cells. Antisera AP-2 and AP-4 were produced by immunizing rabbits with keyhole limpet haemocyanin coupled with synthetic peptides representing KPI region APP301-316 and A beta region APP670-686 of APP770, respectively. These polyclonal antisera were purified against the corresponding peptide using affinity chromatography. Western blot analysis of homogenates of relatively enriched neuronal and astroglial cultures showed that these antibodies discretely stained bands of proteins in a cell-specific manner. Dot-blot analysis using AP-2, AP-4 and 22C11 antibodies indicated that, in comparison with neurons, cultured astrocytes contained 3-fold greater KPI-containing APP isoform proteins. The amount of total APP proteins, which include both KPI-containing and KPI-lacking APP isoforms, was approximately 90% higher in astrocytes than in neurons. Consistent with these in vitro findings in cultured astrocytes, in fimbria-fornix lesioned rat hippocampus, labelling with AP-2 antibody, which specifically reacts with KPI-containing APP proteins, was mainly observed in glial fibrillary acidic protein-positive reactive astrocytes in vivo. The results showed that APP isoforms are expressed in a cell type-specific manner in the brain and, since deposition of A beta is closely associated with the expression of KPI-containing APP isoforms, provide further evidence for the involvement of astrocytes in plaque biogenesis.

Amyloid beta-Protein Precursor↗
PubMed1996

Modulation of N-methyl-D-aspartate receptor mediated arachidonic acid release by endogenous nitric oxide in cultured neurons.

Nitric oxide (NO) and arachidonic acid (AA) are believed to act as intra- and intercellular messengers in the central nervous system. Using primary cultures of striatal and hippocampal neurons, we have studied the effect of endogenous NO on N-methyl-D-aspartate (NMDA)-mediated AA release. Inhibition of NO synthesis with L-NG-nitroarginine (L-Noarg) produced a dose-dependent increase in NMDA-mediated AA release that was reversed by L-arginine. On the other hand, L-Noarg did not modify AA release produced by joint stimulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and phospholipase C-coupled metabotropic subtypes of glutamate receptors. Our results indicate that endogenously produced NO could modulate cellular events mediated by NMDA-induced AA release.

Animals↗
PubMed1996

beta-Amyloid-mediated inhibition of redox activity (MTT reduction) is not an indicator of astroglial degeneration.

The reduction in 3-[4,5-dimethylthiazol]-2,5-diphenyltetrazolium bromide (MTT) to a coloured formazan compound by cultured cells has been extensively used as an in vitro model for understanding neurobiological mechanisms involved in amyloid beta-protein (A beta-mediated cell death. In primary cultures of astrocytes, very low concentrations of aggregated A beta 1-4C, but not A beta 4C-1, produced a significant inhibition in the reduction of the dye MTT. This inhibitory response was rapid and persisted as long as A beta 1-4C was present in the culture medium. Such a severe reduction in cell redox activity for days failed to cause death of astroglial cells, measured in terms of trypan blue uptake and lactate dehydrogenase release. Interleukin-1 beta (IL-1 beta), which is known to attenuate excitotoxic neurodegeneration, had no effect on A beta 1-4C-induced inhibition of MTT reduction. These results suggest that even though inhibition of MTT reduction represents an early indicator of the A beta 1-4C mediated cell injury, without other corroborating evidence, it should not be used as a measure of cell death.

Amyloid beta-Protein Precursor↗
PubMed1996

Glial cell derived neurotrophic factors and Alzheimer's disease.

In Alzheimer's disease the normal balance of metabolic pathways regulating trophic factors/cytokines is disrupted; local reduction may result in neurons being deprived of neurotrophic factors while an excess may initiate a cascade of interaction between glial cells and beta-amyloid precursor protein metabolism thereby facilitating plaque formation. This paper briefly discusses the findings of our group on aspects ranging from cholinergic humoral and trophic factors to mechanisms underlying amyloidogenesis in Alzheimer's disease.

Alzheimer Disease↗
PubMed1995

Neurodegeneration mediated by glutamate and beta-amyloid peptide: a comparison and possible interaction.

In Alzheimer's disease, abnormal extracellular accumulations of beta-amyloid (a major component of the senile plaques) and of the excitatory amino acid glutamate are both thought to be associated with degeneration of nerve cells. In the present study, using cultured cortical or hippocampal neurons as an in vitro model, we compared the effects of various factors influencing neurodegeneration mediated by glutamate or by beta-amyloid peptide (A beta). We also asked the question: does long-term treatment with sublethal doses of A beta-(25-35) potentiate glutamate-mediated excitotoxicity? Neuronal cell death was quantified using the lactate dehydrogenase (LDH) method. Since extracellular LDH remains stable for days, the magnitude of relative afflux of LDH correlates in a linear fashion with the number of damaged neurons in cultures. When applied singly, both glutamate (for 15 min) and A beta-(25-35) or its parent peptide A beta-(1-40) (continuously) produced a dose-dependent neuronal degeneration. In the case of glutamate, the half-maximal effects were observed at about 0.08 mM glutamate for both cerebral cortical and hippocampal neurons (cultured for 13 days in vitro, DIV). The effect of A beta-(25-35) was also time-dependent, while neurons grown in a chemically defined medium showed relatively greater susceptibility to A beta-(25-35) than those cultured in a serum-containing medium. These differential effects were not related to the presence of different numbers of glial cells in the cultures. Treatment with different doses of the antimitotic inhibitor, cytosine arabinoside, for 24 h (6-7 DIV) produced at 13 DIV cortical neuronal cultures with varying numbers of astrocytes, as determined by the astrocyte-specific enzyme glutamine synthetase. The presence of astrocytes decreased the toxicity of glutamate for neurons. The modulation was due to uptake of glutamate by astrocytes, thereby reducing its effective concentration, as the effect was seen at 0.1 mM and not at 10 mM glutamate. Incorporation of an NMDA receptor mediated Ca2+ ion channel blocker, MK-801, together with glutamate completely inhibited degeneration of cortical neurons, and pretreatment of cultures with basic fibroblast growth factor for 2 days did so partially. However, these compounds had no effect on neurotoxicity mediated by A beta-(25-35). Lastly, the effect of glutamate interacted with that of A beta-(25-35). Pretreatment of cortical neurons for 2 days with 10 microM A beta-(25-35) by itself had no appreciable effect, but it potentiated significantly the degeneration of these neurons mediated by glutamate.(ABSTRACT TRUNCATED AT 400 WORDS)

Alzheimer Disease↗
PubMed1995

Effects of protein kinase C modulation on NMDA receptor mediated regulation of neurotransmitter enzyme and c-fos protein in cultured neurons.

The role of protein kinase C (PKC) in N-methyl-D-aspartate (NMDA) receptor-mediated biochemical differentiation and c-fos protein expression was investigated in cultured cerebellar granule neurons. The biochemical differentiation of glutamatergic granule cells was studied in terms of the specific activity of phosphate-activated glutaminase, an enzyme treatment in the synthesis of the putative neurotransmitter pool of glutamate. When the partially depolarized cells were treated with NMDA for the last 1 to 3 days (between 2 and 5 days in vitro), it elevated the specific activity of glutaminase. In contrast, NMDA had little effect on the activity of aspartate aminotransferase or of lactate dehydrogenase. Treatment of 10-day old granule neurons with NMDA also resulted in a marked increase in the immunocytochemically measured expression of c-fos protein. The increases in both the activity of glutaminase and the steady state level of c-fos protein were specific to the activation of NMDA receptors, as they were completely blocked by D,L-2-amino-5-phosphonovaleric acid. The specific stimulation of NMDA receptors in PKC-depleted granule neurons or in the presence of reasonably specific PKC inhibitors also produced significant elevation in the activity of glutaminase and the expression of c-fos protein. These increases were similar in magnitude to those observed in the granule neurons of the respective control groups. Our findings demonstrate that PKC is not directly involved in the NMDA receptor-mediated signal transduction processes associated with biochemical differentiation and c-fos induction in cerebellar granule neurons.

Animals↗
PubMed1995

v-Abl activates c-myc transcription through the E2F site.

The v-abl oncogene of Abelson murine leukemia virus encodes a deregulated form of the cellular nonreceptor tyrosine kinase. v-Abl activates c-myc transcription, and c-Myc is an essential downstream component in the v-Abl transformation program. To explore the mechanism by which v-Abl activates c-myc transcription, a cotransfection assay was developed. We show that transactivation of a c-myc promoter by v-Abl requires the SH1 (tyrosine kinase) and SH2 domains of v-Abl; the C-terminal domains are not required for transactivation. The assay also identified the E2F site in the c-myc promoter as a v-Abl-responsive element. In addition, multimerized E2F sites were shown to be sufficient to confer v-Abl-dependent activation on a minimal promoter. This is the first identification of a v-Abl response element for transcriptional activation. v-Abl tyrosine kinase-dependent changes in proteins binding the c-myc E2F site were also demonstrated, including induction of a complex containing DP1, p107, cyclin A, and cdk2. Identification of v-Abl-dependent changes in E2F-binding proteins provides an important link between v-Abl, transcription, cell cycle regulation, and control of cellular growth.

Abelson murine leukemia virus↗
PubMed1994

MAZ-dependent termination between closely spaced human complement genes.

The zinc finger protein MAZ, originally identified as a factor that binds to the c-myc P2 promoter, is associated with transcriptional termination. As shown in these studies, a termination sequence between the closely spaced human complement genes C2 and Factor B contains a protein binding site which interacts with three different proteins in vitro. Binding of one of these factors, MAZ, correlates with activity of the C2 termination sequence in vivo. Cloned MAZ was used to obtain a consensus binding site, G5AG5. This allowed identification of new sites, between the closely spaced human genes g11 and C4 and within an intron of the mouse IgM-D gene, where termination is known to occur and regulate the expression of IgD. The g11 and IgM MAZ sites lie within sequences that have activity in a termination assay and, furthermore, mutation of C2 or g11 MAZ sites severely reduces termination activity. MAZ bends DNA, and inherently bent DNA is highly active as a terminator, suggesting that MAZ-induced bending is important for C2 and g11 termination. We propose that MAZ sites exist in promoters which require protection against transcriptional interference, such as those of closely spaced genes, to cause efficient termination. The MAZ consensus sequence will facilitate the identification of further sites.

Base Sequence↗
PubMed1993

Colocalization of NGF receptor with VIP in rat suprachiasmatic neurones.

Low-affinity nerve growth factor receptor (p75NGFR) and vasoactive intestinal polypeptide (VIP) immunoreactive neuronal structures and their interrelationship were investigated at light and electron microscopic levels in rat suprachiasmatic nucleus (SCN). p75NGFR immunoreactive neuronal perikarya were detected in the ventrolateral part, while the dorsolateral region contained mainly receptor positive fibres. Double-label immunocytochemistry showed that nearly all p75NGFR positive neurones in the ventrolateral region of the SCN also contained VIP. The axons of the receptor positive neurones terminated on unidentified neurones. In single-label experiments, the axons of the VIP containing neurones formed axodendritic synapses on cells containing the same peptide. The findings provide further insight on the chemical structural organization of the SCN.

Animals↗
PubMed1993

Pseudoexstrophy.

Pseudoexstrophy is a rare, mild exstrophy variant which involves the major musculoskeletal defects of the exstrophy complex without any associated defect in the urinary system. A case is reported presenting at birth as an umbilical positional anomaly. Differential diagnosis and management are reviewed.

Bladder Exstrophy↗
PubMed1993

Regulation of beta-amyloid precursor protein isoform mRNAs by transforming growth factor-beta 1 and interleukin-1 beta in astrocytes.

In cultured astrocytes, all three major transcripts of beta-amyloid precursor protein (APP) were expressed with the ratio for APP695, APP751 and APP770 isoform mRNAs being 1:4:2. In comparison with controls, treatment of astrocytes with transforming growth factor-beta 1 (TGF-beta 1) produced about 6 fold increase in total APP mRNA, while elevation in the interleukin-1 beta (IL-1 beta) treated group was small and may relate to the mitogenic effect of IL-1 beta on astrocytes. Treatment of astrocytes with cytokines also produced marked changes in the upregulation in expression of different APP isoforms. The net increase in mRNAs of KPI-containing isoforms APP751 and APP770 was relatively more than for the APP695 isoform. This phenomenon was mainly related to the differences in the expression of KPI-containing APP isoforms and APP695 isoform in the controls. The present findings provide further evidence for the involvement of astrocytes in a cascade of events leading to the development of senile plaques in Alzheimer's disease and Down's syndrome.

Alzheimer Disease↗
PubMed1993

A low-affinity nerve growth factor receptor antibody is internalized and retrogradely transported selectively into cholinergic neurons of the rat basal forebrain.

The mechanism by which nerve growth factor transduces its signal in responsive cells is yet to be clearly defined. However, it has been suggested that the internalization of nerve growth factor, the first step in the retrograde flow of nerve growth factor, is a property of the high-affinity receptors, p140trkA. Here we show that when a monoclonal antibody (MC 192), which immunoprecipitates p75NGFR (the low-affinity 75,000 mol. wt nerve growth factor receptor protein) and not p140trkA, was administered into the dorsal hippocampal formation of the rats, it was internalized and retrogradely transported to the cell bodies residing in the medial septum-diagonal band complex. The topographic organization and the localization of these neurons containing retrogradely transported p75NGFR antibody were strikingly similar to those nerve cells immunostained for choline acetyltransferase in the immediately-adjacent section, indicating that the neurons which contained p75NGFR antibody were cholinergic neurons. A double-label immunocytochemistry confirmed this conclusion. On the other hand, none of the parvalbumin-positive GABAergic neurons contained retrogradely transported p75NGFR antibody. Moreover, in contrast to specific transport of p75NGFR antibody into cholinergic neurons, when wheat germ agglutinin-colloidal gold was injected into the hippocampus at the same levels, it was taken up and retrogradely transported into both choline acetyltransferase-positive cholinergic and parvalbumin-immunoreactive GABAergic neurons in the medial septum-diagonal band complex.

Animals↗
PubMed1993

Induction of beta-amyloid precursor protein isoform mRNAs by bFGF in astrocytes.

All three major transcripts of beta-amyloid precursor protein (APP) were expressed in astrocytes. The ratio for APP695, APP751 and APP770 isoform mRNAs being 1:4:2. Treatment with bFGF and EGF produced upregulation of APP mRNAs in cultured astrocytes in varying proportions. In comparison with controls, increases in APP mRNAs in bFGF and EGF treated groups were about 4.5 and 2 fold, respectively. The elevation in mRNAs of KPI-containing isoforms APP751 and APP770 was relatively more than the elevation in mRNA of APP695 isoform. Our findings provide further evidence for the involvement of astrocytes in a cascade of events leading to the development of senile plaques in Alzheimer's disease and Down's syndrome.

Amyloid beta-Protein Precursor↗
PubMed1992

Regulation of neurotransmitter enzyme by quisqualate subtype glutamate receptors in cultured cerebellar and hippocampal neurons.

The possible involvement of ionotropic and metabotropic quisqualate (QA) receptors in neuronal plasticity was studied in cultured glutamatergic cerebellar or hippocampal cells in terms of the specific activity of phosphate-activated glutaminase, an enzyme important in the synthesis of the putative neurotransmitter pool of glutamate. When cerebellar or hippocampal neurons were treated with QA, it elevated the specific activity of glutaminase in a dose-dependent manner. The half-maximal effect was obtained at about 0.1 microM, the maximum increase was at about 1 microM, but levels higher than 10 microM QA produced progressive reduction in glutaminase activity. In contrast, QA had little effects on the activities of lactate dehydrogenase and aspartate aminotransferase and the amount of protein, indicating that the increase in glutaminase was relatively specific. The QA-mediated increase in glutaminase was mimicked by the ionotropic QA receptor agonist alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA; EC50, about 0.5 microM), but not by the metabotropic QA receptor agonist trans-(+-)-1-amino-cyclopentyl-1,3,dicarboxylate (t-ACPD; up to 0.5 mM). The specific ionotropic QA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) inhibited QA- and AMPA-mediated increases in glutaminase activity in a dose-dependent manner, whereas other glutamate receptor antagonists, D,L-2-amino-5-phosphonovalerate, gamma-D-glutamyl aminomethyl sulphonic acid and gamma-D-glutamyl diethyl ester were ineffective. The elevation of neurotransmitter enzyme was Ca(2+)-dependent.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗