Jose Aguilar - Academia.edu (original) (raw)
Papers by Jose Aguilar
Neuroscience Letters, Dec 1, 1981
The binding of [3H]L-quinuclidinylbenzilate to muscarinic receptors and [3H]flunitrazepam to benz... more The binding of [3H]L-quinuclidinylbenzilate to muscarinic receptors and [3H]flunitrazepam to benzodiazepine receptors were studied in the rat hippocampus after lesion of the fimbria-fornix. While the muscarinic receptors showed no change, the benzodiazepine receptors did change considerably at various time intervals. Two days after the lesion the specific [3H]flunitrazepam binding decreased 380, while at 5 days it increased 65%. After 14 days of the lesion it still was significantly above normal. These changes are due to a variation in the number of sites, and not in affinity. Possible interpretations of these results are discussed.
European Journal of Pharmacology, Aug 1, 1979
3H-Quinuclidinyl benzylate and dimethyl 14C-d-tubocurarine were used to localize muscarinic and n... more 3H-Quinuclidinyl benzylate and dimethyl 14C-d-tubocurarine were used to localize muscarinic and nicotinic receptors in synaptosomal membranes of the rat cerebral cortex. The results obtained, after the action of different concentrations of Triton X-100, suggest that, while nicotinic receptors are postsynaptic, the muscarinic ones are both pre-and postsynaptic. Receptor localization Synaptic complexes Synaptosomai membranes 3H-QNB binding 14C-DMTC binding
The FASEB Journal, Aug 1, 1989
This article describes ways in which receptors, key components of signal propagation through a sy... more This article describes ways in which receptors, key components of signal propagation through a synapse, can mediate changes in that propagation. Changes occur at four levels: in the signal-transducing capability of a single receptor molecule, in the number of receptors per cell, in the subcellular placement of receptor molecules, and in the cytoarchitecture of receptor-rich regions. The ability of receptors to shift betewen different desired states is called plasticity, and such shifts can be long-lived as well as transient. In this article we focus on neuronal receptors, although key findings from a variety of cell systems are reported. Neuronal receptor plasticity may have a special role in the assembly as well as the adaptability of the nervous system.
Virus Genes, Jun 1, 2007
Herpes simplex virus type 1 (HSV-1) uses multicomponent mechanisms for binding, penetration, and ... more Herpes simplex virus type 1 (HSV-1) uses multicomponent mechanisms for binding, penetration, and cell-to-cell passage. These processes are affected by polysulfonated compounds. In this paper we have addressed the question of whether the same or different interactions of HSV-1 with polysulfonated compounds are involved in binding, penetration, and passage. For this, we have compared the inhibitory dose-response for a series of polysulfonated and cationic compounds known to block HSV-1 infections. These comparisons were done at the level of binding, penetration, and cell-to-cell passage. Variations in the parameters of the dose-response curves - IC(50) and Hill coefficients (n (H)) - are consistent with HSV-1 having multiple interactions with sulfonated cellular components in all these processes. Some of the interactions seem to be common to the three processes, while others are particular for each one.
Journal of Neurochemistry, Aug 1, 1982
ABSTRACT
European Journal of Pharmacology, May 1, 1980
Synaptosomal membranes from cat cerebral cortex were labelled with [3H]-quinuclidinyl benzylate (... more Synaptosomal membranes from cat cerebral cortex were labelled with [3H]-quinuclidinyl benzylate ([3H]-QNB). There was shown to be a single type of binding site with Kd = 0.34 nM, Bmax = 2.2 nmol/g protein and Hill No. = 1.01. Triton X-100 at 5 × 10-4 % inhibited the specific binding of [3H]QNB and the inhibition was almost complete at 10-2 %. Treatment with 2.5 × 10-6 M atropine, followed by centrifugation washings protected the receptor site from the inhibitory action of the detergent. The protection afforded by other cholinergic drugs was less effective. The use of this technique has confirmed the results of our previous work on the possible pre-and postsynaptic location of central muscarinic receptors. These findings open the possibility for protection of other detergent-sensitive receptors and for their isolation and purification as well-defined macromolecules.
Cancer Research, Apr 4, 2023
Chemoresistance is a major therapeutic challenge to prostate cancer and its underlying molecular ... more Chemoresistance is a major therapeutic challenge to prostate cancer and its underlying molecular mechanism is poorly understood. Previously, it has been suggested that bone morphogenetic protein (BMP) signaling is down-regulated during the prostate cancer progression from the early androgen-sensitive stage to the metastatic castration-resistant stage. However, no literature reports are available for BMP signaling in the more advanced-chemoresistant prostate cancer. In this study, we found the expression levels of the BMP type I receptor members, Activin-like kinase-2 (ALK2) and Activin-like kinase-3 (ALK3), were significantly higher in the chemoresistant prostate cancer cells than those in the chemosensitive prostate cancer cells. In addition, the phospho-Smad1/5/9 proteins, the pivotal intracellular effectors of the BMP signaling, were notably elevated in the chemoresistant prostate cancer cells over the chemosensitive prostate cancer cells, indicating that BMP signaling is highly activated in the chemoresistant prostate cancer cells. We also found that BMP signaling inhibition with either DMH1 or the knockdown of ALK2/ALK3 sensitized chemoresistant prostate cancer cells to the chemotherapy drug docetaxel in a dose dependent manner. Our further study indicates that DMH1 suppressed the migration and invasion of chemoresistant prostate cancer cells in vitro, and attenuated chemoresistant prostate tumor growth in the mouse xenograft model in vivo. In addition, we showed that DMH1 disrupted the sphere formation in DU145-TxR and PC3-TxR cells, suppressed the expression of marker genes of the cancer stem cells (CSCs), and inhibited the efflux activity of multidrug resistance P-glycoprotein (P-gp1). In conclusion, our study demonstrates that BMP signaling is associated with prostate cancer chemoresistance and BMP signaling inhibition effectively overcomes the cancer chemoresistance potentially through the disruption of CSCs’ stemness and P-gp1 efflux activity. Citation Format: Jijun Hao, Chen Xie, Zhijun Wang, Yong Bar, Jose Aguilar, Austin Kyan, Li Zhing. BMP inhibition overcomes chemoresistance of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2844.
Humana Press eBooks, Oct 26, 2004
The design and construction of a long (75-mer) oligonucleotide-based DNA microarray for herpes si... more The design and construction of a long (75-mer) oligonucleotide-based DNA microarray for herpes simplex virus type 2 transcripts is described. This array is utilized to generate an analysis of HSV-2 transcript abundance as a function of conditions of infection of human cells, and global patterns of HSV-2 transcript abundance are compared with those for HSV-1. General similarities in patterns along with notable differences in specific details are noted. These results reveal a marked conservation in the program of gene activity between phenotypically diverged strains.
Journal Of Receptors And Signal Transduction, 1981
Membranes were isolated from the cerebral cortices of rat brain and submitted to binding with 3H-... more Membranes were isolated from the cerebral cortices of rat brain and submitted to binding with 3H-flunitrazepam (3H-FNZP). Specific binding gave a hyperbolic saturation curve reaching a maximum between 10 - 12 nM suggesting a single population of sites. By Scatchard analysis a KD = 4 nM and a Bmax = 407 fmol/mg protein was obtained; the Hill number was 0.97. The effect of Triton X-100 was analyzed between concentrations of 6.4 x 10(-8) and 5 x 10(-1)%. With 6.4 x 10(-6) and 6.4 x 10(-3)% there is an activation of the binding without loss of protein. This activation is not reversed by washing. At 6.4 x 10(-2)% or higher concentrations there is an inhibition that is partially reversed and also solubilization of some receptors. The activation with low detergent is due to an increase in Bmax without change in KD. With 2 x 10(-1)% and 5 x 10(-1)% Triton X-100 there is considerable decrease in Bmax and in th latter case, an increase in affinity, as well. The results obtained through the action of Triton X-100 suggest that a proportion of benzodiazepine receptors are localized presynaptically. There findings are discussed in relation to previous studies from this laboratory on the localization of other central receptors with reference to the synaptic region.
Journal of NeuroVirology, Dec 1, 2009
Herpes simplex virus type 1 (HSV-1)-based vectors readily transduce neurons and have a large payl... more Herpes simplex virus type 1 (HSV-1)-based vectors readily transduce neurons and have a large payload capacity, making them particularly amenable to gene therapy applications within the central nervous system (CNS). Because aspects of the host responses to HSV-1 vectors in the CNS are largely unknown, we compared the host response of a nonreplicating HSV-1 vector to that of a replicationcompetent HSV-1 virus using microarray analysis. In parallel, HSV-1 gene expression was tracked using HSV-specific oligonucleotide-based arrays in order to correlate viral gene expression with observed changes in host response. Microarray analysis was performed following stereotactic injection into the right hippocampal formation of mice with either a replication-competent HSV-1 or a nonreplicating recombinant of HSV-1, lacking the ICP4 gene (ICP4−). Genes that demonstrated a significant change (P < .001) in expression in response to the replicating HSV-1 outnumbered those that changed in response to mock or nonreplicating vector by approximately 3-fold. Pathway analysis revealed that both the replicating and nonreplicating vectors induced robust antigen presentation but only mild interferon, chemokine, and cytokine signaling responses. The ICP4− vector was restricted in several of the Toll-like receptor-signaling pathways, indicating reduced stimulation of the innate immune response. These array analyses suggest that although the nonreplicating vector induces detectable activation of immune response pathways, the number and magnitude of the induced response is dramatically restricted compared to the replicating vector, and with the exception of antigen presentation, host gene expression induced by the non-replicating vector largely resembles mock infection.
World Neurosurgery, Jul 1, 2018
Neurochemical Research, Dec 1, 1992
Catecholamine secretion in the bovine adrenal medulla is evoked largely by nicotinic receptor act... more Catecholamine secretion in the bovine adrenal medulla is evoked largely by nicotinic receptor activation. However, bovine adrenal medulla also contain muscarinic receptors that mediate several cell responses. To understand the physiological role of muscarinic receptors in the bovine adrenal medulla it is important to identify the pharmacological subtypes present in this tissue. For this, we analyzed the abilities of different selective muscarinic antagonists in displacing the binding of the non-selective antagonist [3H] quinuclidinyl benzylate to an enriched plasma membrane fraction prepared from bovine adrenal medulla. All the selective antagonists bind at least two bindings sites with different affinities. The binding profile of the sites with high proportion is similar to the M2 subtype and those present in low proportion have a M1 profile. However, some variation in the proportion of the sites for the different ligands suggest the presence of the third pharmacological subtype (M3). We conclude that the sites in high proportion (60-80%) correspond to M2 muscarinic subtypes, and the rest is constituted by M1 plus M3 subtypes. The presence of multiplicity of subtypes in the adrenal medulla membranes suggests a diversity of functions of muscarinic receptors in the adrenal gland.
Virology, 2006
The genomes of human herpes virus type-1 and type-2 share a high degree of sequence identity; yet... more The genomes of human herpes virus type-1 and type-2 share a high degree of sequence identity; yet, they exhibit important differences in pathology in their natural human host as well as in animal host and cell cultures. Here, we report the comparative analysis of the time and relative abundance profiles of the transcription of each virus type (their transcriptomes) using parallel infections and microarray analysis using HSV-1 probes which hybridize with high efficiency to orthologous HSV-2 transcripts. We have confirmed that orthologous transcripts belong to the same kinetic class; however, the temporal pattern of accumulation of 4 transcripts (U L 4, U L 29, U L 30, and U L 31) differs in infections between the two virus types. Interestingly, the protein products of these transcripts are all involved in nuclear organization and viral DNA localization. We discuss the relevance of these findings and whether they may have potential roles in the pathological differences of HSV-1 and HSV-2.
Neuroscience Letters, 2001
Our aim was to investigate the neuromodulatory role of diadenosine tetraphosphate (Ap 4 A). Ap 4 ... more Our aim was to investigate the neuromodulatory role of diadenosine tetraphosphate (Ap 4 A). Ap 4 A-binding sites were detected in striatum and hippocampus membranes using [ 35 S]-ADPbS as radioligand and Ap 4 A and 1-(Ap 4 A), di-ethenoadenosine tetraphosphate, as displacers. Effects of 1-(Ap 4 A) on extracellular glutamate levels were studied using intracerebral perfusion. Both areas contain high-af®nity binding sites for [ 35 S]-ADPbS with K d values in the low nM range. [ 35 S]-ADPbS binding was displaced by Ap 4 A and 1-(Ap 4 A). At 1 and 10 mM doses, 1-(Ap 4 A) markedly decreased glutamate levels in the striatum. The possibility of Ap 4 A acting as an endogenous modulator of excitatory neurotransmission is discussed.
Journal of Receptors and Signal Transduction, 1980
AbstractProteolipids (i.e., hydrophobic proteins) have been extracted with chloroform-methanol (2... more AbstractProteolipids (i.e., hydrophobic proteins) have been extracted with chloroform-methanol (2:1) from lyophilized Torpedo electroplax, and fractionated on a DEAE-cellulose column. The elution system consisted of the same solvent and a gradient of the hydrophobic ion ptoluene sulfonate (0.1–100mM). The three fractions obtained (I, II and III) have different content of protein, lipid P, and reducing sugars. The amino acid composition shows a higher proportion of hydrophobic residues in I and more charged ones in fractions II and III. Polyacryl amide gel electrophoresis of fraction I shows a single major band of 39 kdaltons; in fractions II and III a major band of 42 kdaltons and fainter bands in the range 62–68 kdaltons are observed.Fraction I has the highest specific binding for [3H]-acetylcholine (7.1 nmol/mg protein) and [3H]α-bungarotoxin (5.2 nmol/mg protein). The nicotinic nature of this proteolipid was demonstrated by blocking experiments. The Scatchard plot showed two affinity sites for [3H]-ace...
![Research paper thumbnail of Inhibition by local anesthetics, phentolamine and propranolol of [3H]Quinuclydinyl benzylate binding to central muscarinic receptors](https://attachments.academia-assets.com/113873155/thumbnails/1.jpg)
](European Journal of Pharmacology, 1980
317-326. [ 3H ] Quinuclydinyl benzylate ([ 3H ] Q NB) binding was carried out on crude synaptosom... more 317-326. [ 3H ] Quinuclydinyl benzylate ([ 3H ] Q NB) binding was carried out on crude synaptosomal membranes isolated from eat cerebral cortex. The specific binding showed a single type of site with K D 0.25 nM, Hill number 0.89 and Bma x 114 pmol/g protein. The local anesthetics procaine, tetracaine and dibucaine, and the adrenergic antagonists phentolamine and propranolol, in concentrations between 1 nM and 500 pM, inhibited [3H] QNB binding with K i varying between 9 pM for procaine and 80 pM for propranolol. The Hill coefficients obtained from logit/ log plots suggested that there was no cooperativity between the binding sites for local anesthetics. At various concentrations the inhibition by procaine and propranolol may appear as competitive or non-competitive. The Hill numbers obtained from the saturation curves suggest that there was no cooperativity between anesthetics and [3H]QNB binding sites. Neither 1 mM Ca 2+ nor Mg 2+ affected [3H]QNB binding or the action of the drugs. The effect of local anesthetics and adrenergic antagonists was reversible and these drugs did not protect the muscarinic receptor from the deleterious effect of Triton X-100 as was the case with muscarinic agents. Our findings suggest that local anesthetics inhibit [ ~H]QNB binding to the muscarinic receptor by acting at some accessory site but not on the true receptor site. The possible mechanism of action of local anesthetics on synaptic transmission is discussed.
Brain Research, 1982
Bilateral intraventricular injections of 0.5 pg of kai,ic acid were used to selectively destroy C... more Bilateral intraventricular injections of 0.5 pg of kai,ic acid were used to selectively destroy CA3 hippocampal pyramidal neurons, in an effort to clarify the possible localization of muscarinic cholinergic receptors in the rat hippocampal formation. Thirty days after treatment, there was a 43 % decrease in the total number of [aH]L-QNB binding sites per hippocampus, wifi, no change in affinity. Histological examination confirmed the selective loss of pyra~:-;~al neurons in subareas CA3a-b while other regions of the hippocampal formation were spared. The unilateral transection of the fimbria-fornix, done 14 days after kainic acid, produced a further reduction in binding in relation to control hippocampi (-57 ~o). The results demonstrate that in the pyramidal cells of CA3 there is a high concentration of postsynaptic muscarinic receptors. However, the slight further decrease, found after fimbria-fornix transection, suggests the possible existeqce of a small population of presynaptic receptors that, hitherto, had only been demonstrated indirectly by physiological methods.
Neuroscience Letters, Dec 1, 1981
The binding of [3H]L-quinuclidinylbenzilate to muscarinic receptors and [3H]flunitrazepam to benz... more The binding of [3H]L-quinuclidinylbenzilate to muscarinic receptors and [3H]flunitrazepam to benzodiazepine receptors were studied in the rat hippocampus after lesion of the fimbria-fornix. While the muscarinic receptors showed no change, the benzodiazepine receptors did change considerably at various time intervals. Two days after the lesion the specific [3H]flunitrazepam binding decreased 380, while at 5 days it increased 65%. After 14 days of the lesion it still was significantly above normal. These changes are due to a variation in the number of sites, and not in affinity. Possible interpretations of these results are discussed.
European Journal of Pharmacology, Aug 1, 1979
3H-Quinuclidinyl benzylate and dimethyl 14C-d-tubocurarine were used to localize muscarinic and n... more 3H-Quinuclidinyl benzylate and dimethyl 14C-d-tubocurarine were used to localize muscarinic and nicotinic receptors in synaptosomal membranes of the rat cerebral cortex. The results obtained, after the action of different concentrations of Triton X-100, suggest that, while nicotinic receptors are postsynaptic, the muscarinic ones are both pre-and postsynaptic. Receptor localization Synaptic complexes Synaptosomai membranes 3H-QNB binding 14C-DMTC binding
The FASEB Journal, Aug 1, 1989
This article describes ways in which receptors, key components of signal propagation through a sy... more This article describes ways in which receptors, key components of signal propagation through a synapse, can mediate changes in that propagation. Changes occur at four levels: in the signal-transducing capability of a single receptor molecule, in the number of receptors per cell, in the subcellular placement of receptor molecules, and in the cytoarchitecture of receptor-rich regions. The ability of receptors to shift betewen different desired states is called plasticity, and such shifts can be long-lived as well as transient. In this article we focus on neuronal receptors, although key findings from a variety of cell systems are reported. Neuronal receptor plasticity may have a special role in the assembly as well as the adaptability of the nervous system.
Virus Genes, Jun 1, 2007
Herpes simplex virus type 1 (HSV-1) uses multicomponent mechanisms for binding, penetration, and ... more Herpes simplex virus type 1 (HSV-1) uses multicomponent mechanisms for binding, penetration, and cell-to-cell passage. These processes are affected by polysulfonated compounds. In this paper we have addressed the question of whether the same or different interactions of HSV-1 with polysulfonated compounds are involved in binding, penetration, and passage. For this, we have compared the inhibitory dose-response for a series of polysulfonated and cationic compounds known to block HSV-1 infections. These comparisons were done at the level of binding, penetration, and cell-to-cell passage. Variations in the parameters of the dose-response curves - IC(50) and Hill coefficients (n (H)) - are consistent with HSV-1 having multiple interactions with sulfonated cellular components in all these processes. Some of the interactions seem to be common to the three processes, while others are particular for each one.
Journal of Neurochemistry, Aug 1, 1982
ABSTRACT
European Journal of Pharmacology, May 1, 1980
Synaptosomal membranes from cat cerebral cortex were labelled with [3H]-quinuclidinyl benzylate (... more Synaptosomal membranes from cat cerebral cortex were labelled with [3H]-quinuclidinyl benzylate ([3H]-QNB). There was shown to be a single type of binding site with Kd = 0.34 nM, Bmax = 2.2 nmol/g protein and Hill No. = 1.01. Triton X-100 at 5 × 10-4 % inhibited the specific binding of [3H]QNB and the inhibition was almost complete at 10-2 %. Treatment with 2.5 × 10-6 M atropine, followed by centrifugation washings protected the receptor site from the inhibitory action of the detergent. The protection afforded by other cholinergic drugs was less effective. The use of this technique has confirmed the results of our previous work on the possible pre-and postsynaptic location of central muscarinic receptors. These findings open the possibility for protection of other detergent-sensitive receptors and for their isolation and purification as well-defined macromolecules.
Cancer Research, Apr 4, 2023
Chemoresistance is a major therapeutic challenge to prostate cancer and its underlying molecular ... more Chemoresistance is a major therapeutic challenge to prostate cancer and its underlying molecular mechanism is poorly understood. Previously, it has been suggested that bone morphogenetic protein (BMP) signaling is down-regulated during the prostate cancer progression from the early androgen-sensitive stage to the metastatic castration-resistant stage. However, no literature reports are available for BMP signaling in the more advanced-chemoresistant prostate cancer. In this study, we found the expression levels of the BMP type I receptor members, Activin-like kinase-2 (ALK2) and Activin-like kinase-3 (ALK3), were significantly higher in the chemoresistant prostate cancer cells than those in the chemosensitive prostate cancer cells. In addition, the phospho-Smad1/5/9 proteins, the pivotal intracellular effectors of the BMP signaling, were notably elevated in the chemoresistant prostate cancer cells over the chemosensitive prostate cancer cells, indicating that BMP signaling is highly activated in the chemoresistant prostate cancer cells. We also found that BMP signaling inhibition with either DMH1 or the knockdown of ALK2/ALK3 sensitized chemoresistant prostate cancer cells to the chemotherapy drug docetaxel in a dose dependent manner. Our further study indicates that DMH1 suppressed the migration and invasion of chemoresistant prostate cancer cells in vitro, and attenuated chemoresistant prostate tumor growth in the mouse xenograft model in vivo. In addition, we showed that DMH1 disrupted the sphere formation in DU145-TxR and PC3-TxR cells, suppressed the expression of marker genes of the cancer stem cells (CSCs), and inhibited the efflux activity of multidrug resistance P-glycoprotein (P-gp1). In conclusion, our study demonstrates that BMP signaling is associated with prostate cancer chemoresistance and BMP signaling inhibition effectively overcomes the cancer chemoresistance potentially through the disruption of CSCs’ stemness and P-gp1 efflux activity. Citation Format: Jijun Hao, Chen Xie, Zhijun Wang, Yong Bar, Jose Aguilar, Austin Kyan, Li Zhing. BMP inhibition overcomes chemoresistance of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2844.
Humana Press eBooks, Oct 26, 2004
The design and construction of a long (75-mer) oligonucleotide-based DNA microarray for herpes si... more The design and construction of a long (75-mer) oligonucleotide-based DNA microarray for herpes simplex virus type 2 transcripts is described. This array is utilized to generate an analysis of HSV-2 transcript abundance as a function of conditions of infection of human cells, and global patterns of HSV-2 transcript abundance are compared with those for HSV-1. General similarities in patterns along with notable differences in specific details are noted. These results reveal a marked conservation in the program of gene activity between phenotypically diverged strains.
Journal Of Receptors And Signal Transduction, 1981
Membranes were isolated from the cerebral cortices of rat brain and submitted to binding with 3H-... more Membranes were isolated from the cerebral cortices of rat brain and submitted to binding with 3H-flunitrazepam (3H-FNZP). Specific binding gave a hyperbolic saturation curve reaching a maximum between 10 - 12 nM suggesting a single population of sites. By Scatchard analysis a KD = 4 nM and a Bmax = 407 fmol/mg protein was obtained; the Hill number was 0.97. The effect of Triton X-100 was analyzed between concentrations of 6.4 x 10(-8) and 5 x 10(-1)%. With 6.4 x 10(-6) and 6.4 x 10(-3)% there is an activation of the binding without loss of protein. This activation is not reversed by washing. At 6.4 x 10(-2)% or higher concentrations there is an inhibition that is partially reversed and also solubilization of some receptors. The activation with low detergent is due to an increase in Bmax without change in KD. With 2 x 10(-1)% and 5 x 10(-1)% Triton X-100 there is considerable decrease in Bmax and in th latter case, an increase in affinity, as well. The results obtained through the action of Triton X-100 suggest that a proportion of benzodiazepine receptors are localized presynaptically. There findings are discussed in relation to previous studies from this laboratory on the localization of other central receptors with reference to the synaptic region.
Journal of NeuroVirology, Dec 1, 2009
Herpes simplex virus type 1 (HSV-1)-based vectors readily transduce neurons and have a large payl... more Herpes simplex virus type 1 (HSV-1)-based vectors readily transduce neurons and have a large payload capacity, making them particularly amenable to gene therapy applications within the central nervous system (CNS). Because aspects of the host responses to HSV-1 vectors in the CNS are largely unknown, we compared the host response of a nonreplicating HSV-1 vector to that of a replicationcompetent HSV-1 virus using microarray analysis. In parallel, HSV-1 gene expression was tracked using HSV-specific oligonucleotide-based arrays in order to correlate viral gene expression with observed changes in host response. Microarray analysis was performed following stereotactic injection into the right hippocampal formation of mice with either a replication-competent HSV-1 or a nonreplicating recombinant of HSV-1, lacking the ICP4 gene (ICP4−). Genes that demonstrated a significant change (P < .001) in expression in response to the replicating HSV-1 outnumbered those that changed in response to mock or nonreplicating vector by approximately 3-fold. Pathway analysis revealed that both the replicating and nonreplicating vectors induced robust antigen presentation but only mild interferon, chemokine, and cytokine signaling responses. The ICP4− vector was restricted in several of the Toll-like receptor-signaling pathways, indicating reduced stimulation of the innate immune response. These array analyses suggest that although the nonreplicating vector induces detectable activation of immune response pathways, the number and magnitude of the induced response is dramatically restricted compared to the replicating vector, and with the exception of antigen presentation, host gene expression induced by the non-replicating vector largely resembles mock infection.
World Neurosurgery, Jul 1, 2018
Neurochemical Research, Dec 1, 1992
Catecholamine secretion in the bovine adrenal medulla is evoked largely by nicotinic receptor act... more Catecholamine secretion in the bovine adrenal medulla is evoked largely by nicotinic receptor activation. However, bovine adrenal medulla also contain muscarinic receptors that mediate several cell responses. To understand the physiological role of muscarinic receptors in the bovine adrenal medulla it is important to identify the pharmacological subtypes present in this tissue. For this, we analyzed the abilities of different selective muscarinic antagonists in displacing the binding of the non-selective antagonist [3H] quinuclidinyl benzylate to an enriched plasma membrane fraction prepared from bovine adrenal medulla. All the selective antagonists bind at least two bindings sites with different affinities. The binding profile of the sites with high proportion is similar to the M2 subtype and those present in low proportion have a M1 profile. However, some variation in the proportion of the sites for the different ligands suggest the presence of the third pharmacological subtype (M3). We conclude that the sites in high proportion (60-80%) correspond to M2 muscarinic subtypes, and the rest is constituted by M1 plus M3 subtypes. The presence of multiplicity of subtypes in the adrenal medulla membranes suggests a diversity of functions of muscarinic receptors in the adrenal gland.
Virology, 2006
The genomes of human herpes virus type-1 and type-2 share a high degree of sequence identity; yet... more The genomes of human herpes virus type-1 and type-2 share a high degree of sequence identity; yet, they exhibit important differences in pathology in their natural human host as well as in animal host and cell cultures. Here, we report the comparative analysis of the time and relative abundance profiles of the transcription of each virus type (their transcriptomes) using parallel infections and microarray analysis using HSV-1 probes which hybridize with high efficiency to orthologous HSV-2 transcripts. We have confirmed that orthologous transcripts belong to the same kinetic class; however, the temporal pattern of accumulation of 4 transcripts (U L 4, U L 29, U L 30, and U L 31) differs in infections between the two virus types. Interestingly, the protein products of these transcripts are all involved in nuclear organization and viral DNA localization. We discuss the relevance of these findings and whether they may have potential roles in the pathological differences of HSV-1 and HSV-2.
Neuroscience Letters, 2001
Our aim was to investigate the neuromodulatory role of diadenosine tetraphosphate (Ap 4 A). Ap 4 ... more Our aim was to investigate the neuromodulatory role of diadenosine tetraphosphate (Ap 4 A). Ap 4 A-binding sites were detected in striatum and hippocampus membranes using [ 35 S]-ADPbS as radioligand and Ap 4 A and 1-(Ap 4 A), di-ethenoadenosine tetraphosphate, as displacers. Effects of 1-(Ap 4 A) on extracellular glutamate levels were studied using intracerebral perfusion. Both areas contain high-af®nity binding sites for [ 35 S]-ADPbS with K d values in the low nM range. [ 35 S]-ADPbS binding was displaced by Ap 4 A and 1-(Ap 4 A). At 1 and 10 mM doses, 1-(Ap 4 A) markedly decreased glutamate levels in the striatum. The possibility of Ap 4 A acting as an endogenous modulator of excitatory neurotransmission is discussed.
Journal of Receptors and Signal Transduction, 1980
AbstractProteolipids (i.e., hydrophobic proteins) have been extracted with chloroform-methanol (2... more AbstractProteolipids (i.e., hydrophobic proteins) have been extracted with chloroform-methanol (2:1) from lyophilized Torpedo electroplax, and fractionated on a DEAE-cellulose column. The elution system consisted of the same solvent and a gradient of the hydrophobic ion ptoluene sulfonate (0.1–100mM). The three fractions obtained (I, II and III) have different content of protein, lipid P, and reducing sugars. The amino acid composition shows a higher proportion of hydrophobic residues in I and more charged ones in fractions II and III. Polyacryl amide gel electrophoresis of fraction I shows a single major band of 39 kdaltons; in fractions II and III a major band of 42 kdaltons and fainter bands in the range 62–68 kdaltons are observed.Fraction I has the highest specific binding for [3H]-acetylcholine (7.1 nmol/mg protein) and [3H]α-bungarotoxin (5.2 nmol/mg protein). The nicotinic nature of this proteolipid was demonstrated by blocking experiments. The Scatchard plot showed two affinity sites for [3H]-ace...
European Journal of Pharmacology, 1980
317-326. [ 3H ] Quinuclydinyl benzylate ([ 3H ] Q NB) binding was carried out on crude synaptosom... more 317-326. [ 3H ] Quinuclydinyl benzylate ([ 3H ] Q NB) binding was carried out on crude synaptosomal membranes isolated from eat cerebral cortex. The specific binding showed a single type of site with K D 0.25 nM, Hill number 0.89 and Bma x 114 pmol/g protein. The local anesthetics procaine, tetracaine and dibucaine, and the adrenergic antagonists phentolamine and propranolol, in concentrations between 1 nM and 500 pM, inhibited [3H] QNB binding with K i varying between 9 pM for procaine and 80 pM for propranolol. The Hill coefficients obtained from logit/ log plots suggested that there was no cooperativity between the binding sites for local anesthetics. At various concentrations the inhibition by procaine and propranolol may appear as competitive or non-competitive. The Hill numbers obtained from the saturation curves suggest that there was no cooperativity between anesthetics and [3H]QNB binding sites. Neither 1 mM Ca 2+ nor Mg 2+ affected [3H]QNB binding or the action of the drugs. The effect of local anesthetics and adrenergic antagonists was reversible and these drugs did not protect the muscarinic receptor from the deleterious effect of Triton X-100 as was the case with muscarinic agents. Our findings suggest that local anesthetics inhibit [ ~H]QNB binding to the muscarinic receptor by acting at some accessory site but not on the true receptor site. The possible mechanism of action of local anesthetics on synaptic transmission is discussed.
Brain Research, 1982
Bilateral intraventricular injections of 0.5 pg of kai,ic acid were used to selectively destroy C... more Bilateral intraventricular injections of 0.5 pg of kai,ic acid were used to selectively destroy CA3 hippocampal pyramidal neurons, in an effort to clarify the possible localization of muscarinic cholinergic receptors in the rat hippocampal formation. Thirty days after treatment, there was a 43 % decrease in the total number of [aH]L-QNB binding sites per hippocampus, wifi, no change in affinity. Histological examination confirmed the selective loss of pyra~:-;~al neurons in subareas CA3a-b while other regions of the hippocampal formation were spared. The unilateral transection of the fimbria-fornix, done 14 days after kainic acid, produced a further reduction in binding in relation to control hippocampi (-57 ~o). The results demonstrate that in the pyramidal cells of CA3 there is a high concentration of postsynaptic muscarinic receptors. However, the slight further decrease, found after fimbria-fornix transection, suggests the possible existeqce of a small population of presynaptic receptors that, hitherto, had only been demonstrated indirectly by physiological methods.