{"id":{"repo_id":"sheffield-hallam","oai_identifier":"oai:shura.shu.ac.uk:19409"},"canonical_url":"https://search.dev.ndltd.org/etd/sheffield-hallam/oai:shura.shu.ac.uk:19409","repository":{"repo_id":"sheffield-hallam","name":"Sheffield Hallam University","base_url":"https://shura.shu.ac.uk/cgi/oai2"},"display":{"title":"5-HT1A receptor expression : Studies in postmortem tissue and characterisation of a model system.","abstract":"Serotonin (5-HT) neurotransmission is involved in the psychopharmacology of several psychiatric disorders including, depression, anxiety disorders and schizophrenia. The release of 5-HT in neurons is mediated by somatodendritic 5-HT[1A] autoreceptors. The presence of 5-HT[1A] receptor is thought to be increased in depressed patients, producing a reduction in the synthesis of 5-HT. A common single nucleotide polymorphism in the promoter region of 5HT[1A] receptor C-1019G is also associated with depression and suicide. The nuclear DEAF-1 related (NUDR) protein represses the 5-HT[1A] promoter region hence regulating both 5-HT[1A] transcription and receptor expression. The project involved undertaking a post-mortem study to determine any association between the 5-HT[1A] promoter polymorphism and the expression of 5-HT[1A] receptor mRNA and receptor density in control human hippocampal brain tissue. This was achieved by genotyping human brain tissue for the 5-HT[1A] receptor polymorphism C-1019G and 5-HT[1A] receptor mRNA levels were quantified using real-time PCR. Radioligand binding was used to determine B[max] and Kd quantifying 5-HT[1A] receptor density. The SHSY-5Y neuroblastoma cell line is a well characterised cell line model used in neurotransmitter studies when differentiated. The 5-HT[1A] receptor couples to Gi proteins inhibiting AC activity and hence mediating a variety of intracellular changes such as, decreasing cAMP leading to decreased Ca[2+] levels. The SH-SY5Y cell line study investigated whether the 5-HT[1A] agonist 8-OH-DPAT, inhibited forskolin stimulated Ca[2+] release in the SHSY-5Y cell line and whether the 5-HT[1A] antagonist p-MPPI reversed this effect using flow cytometry. The post-mortem study showed that the G-1019 allele had significantly higher 5-HT[1A] expression compared to the C allele in control hippocampal tissue. Radioligand binding data demonstrated that control samples with a GG or G/C genotype had a significantly higher 5-HT[1A] receptor density compared to samples with a CC genotype. SH-SY5Y cells differentiated with RA for 5 days or NGF and aphidicolin for 10 days had significantly increased 5-HT 1A receptor mRNA levels compared to undifferentiated cells. Western blots and immunocytochemistry confirmed the presence of the 5-HT[1A] receptor in this cell line. An increase in NUDR expression was observed at the same time there is an increase in 5-HT[1A] receptor expression in SH-SY5Y cells treated with RA or NGF and aphidicolin. Flow cytometry showed that 8-OH-DPAT efficiently diminished forskolin-stimulated increase in intracellular Ca[2+] in RA differentiated cells. 5-HT also a 5-HT[1A] agonist had a similar effect. SH-SY5Y cells treated with both p-MPPI and 8-OH-DPAT demonstrated that cells treated with p-MPPI at higher concentrations significantly increased forskolin-stimulated intracellular Ca[2+] levels and therefore effectively reversed the agonistic effect of 8-OH-DPAT. The findings presented in the post-mortem study are novel and the SH-SY5Y cell line study demonstrates that this cell line when differentiated with either RA or NGF and aphidicolin is a useful cell-line model system for studying the 5-HT[1A] receptor.","abstract_html":"Serotonin (5-HT) neurotransmission is involved in the psychopharmacology of several psychiatric disorders including, depression, anxiety disorders and schizophrenia. The release of 5-HT in neurons is mediated by somatodendritic 5-HT[1A] autoreceptors. The presence of 5-HT[1A] receptor is thought to be increased in depressed patients, producing a reduction in the synthesis of 5-HT. A common single nucleotide polymorphism in the promoter region of 5HT[1A] receptor C-1019G is also associated with depression and suicide. The nuclear DEAF-1 related (NUDR) protein represses the 5-HT[1A] promoter region hence regulating both 5-HT[1A] transcription and receptor expression. The project involved undertaking a post-mortem study to determine any association between the 5-HT[1A] promoter polymorphism and the expression of 5-HT[1A] receptor mRNA and receptor density in control human hippocampal brain tissue. This was achieved by genotyping human brain tissue for the 5-HT[1A] receptor polymorphism C-1019G and 5-HT[1A] receptor mRNA levels were quantified using real-time PCR. Radioligand binding was used to determine B[max] and Kd quantifying 5-HT[1A] receptor density. The SHSY-5Y neuroblastoma cell line is a well characterised cell line model used in neurotransmitter studies when differentiated. The 5-HT[1A] receptor couples to Gi proteins inhibiting AC activity and hence mediating a variety of intracellular changes such as, decreasing cAMP leading to decreased Ca[2+] levels. The SH-SY5Y cell line study investigated whether the 5-HT[1A] agonist 8-OH-DPAT, inhibited forskolin stimulated Ca[2+] release in the SHSY-5Y cell line and whether the 5-HT[1A] antagonist p-MPPI reversed this effect using flow cytometry. The post-mortem study showed that the G-1019 allele had significantly higher 5-HT[1A] expression compared to the C allele in control hippocampal tissue. Radioligand binding data demonstrated that control samples with a GG or G/C genotype had a significantly higher 5-HT[1A] receptor density compared to samples with a CC genotype. SH-SY5Y cells differentiated with RA for 5 days or NGF and aphidicolin for 10 days had significantly increased 5-HT 1A receptor mRNA levels compared to undifferentiated cells. Western blots and immunocytochemistry confirmed the presence of the 5-HT[1A] receptor in this cell line. An increase in NUDR expression was observed at the same time there is an increase in 5-HT[1A] receptor expression in SH-SY5Y cells treated with RA or NGF and aphidicolin. Flow cytometry showed that 8-OH-DPAT efficiently diminished forskolin-stimulated increase in intracellular Ca[2+] in RA differentiated cells. 5-HT also a 5-HT[1A] agonist had a similar effect. SH-SY5Y cells treated with both p-MPPI and 8-OH-DPAT demonstrated that cells treated with p-MPPI at higher concentrations significantly increased forskolin-stimulated intracellular Ca[2+] levels and therefore effectively reversed the agonistic effect of 8-OH-DPAT. The findings presented in the post-mortem study are novel and the SH-SY5Y cell line study demonstrates that this cell line when differentiated with either RA or NGF and aphidicolin is a useful cell-line model system for studying the 5-HT[1A] receptor.","abstract_has_math":false,"creators":["Bunn, Lindsey J."],"institution":"Sheffield Hallam University (United Kingdom).","degree_name":"phd","degree_level":"doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Dalton, Caroline","Hall, Adrian","Reynolds, Gavin"],"committee_chairs":[],"committee_members":[],"year":2008,"date_issued":"2008","date_published":"2008","updated_at":"2026-07-24T06:31:14Z","subjects":[],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Dalton, Caroline","Hall, Adrian","Reynolds, Gavin"]},{"key":"dc:creator","label":"Author","values":["Bunn, Lindsey J."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2008"]},{"key":"dc:date.issued","label":"Date","values":["2008"]},{"key":"dc:publisher.commercial","label":"Dc Publisher Commercial","values":["Sheffield Hallam University,"]},{"key":"dc:publisher.department","label":"Dc Publisher Department","values":["Department Not Provided."]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["Sheffield Hallam University (United Kingdom)."]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://shura.shu.ac.uk/19409/"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["doctoral"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["phd"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://shura.shu.ac.uk/19409/1/10694290.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Serotonin (5-HT) neurotransmission is involved in the psychopharmacology of several psychiatric disorders including, depression, anxiety disorders and schizophrenia. The release of 5-HT in neurons is mediated by somatodendritic 5-HT[1A] autoreceptors. The presence of 5-HT[1A] receptor is thought to be increased in depressed patients, producing a reduction in the synthesis of 5-HT. A common single nucleotide polymorphism in the promoter region of 5HT[1A] receptor C-1019G is also associated with depression and suicide. The nuclear DEAF-1 related (NUDR) protein represses the 5-HT[1A] promoter region hence regulating both 5-HT[1A] transcription and receptor expression. The project involved undertaking a post-mortem study to determine any association between the 5-HT[1A] promoter polymorphism and the expression of 5-HT[1A] receptor mRNA and receptor density in control human hippocampal brain tissue. This was achieved by genotyping human brain tissue for the 5-HT[1A] receptor polymorphism C-1019G and 5-HT[1A] receptor mRNA levels were quantified using real-time PCR. Radioligand binding was used to determine B[max] and Kd quantifying 5-HT[1A] receptor density. The SHSY-5Y neuroblastoma cell line is a well characterised cell line model used in neurotransmitter studies when differentiated. The 5-HT[1A] receptor couples to Gi proteins inhibiting AC activity and hence mediating a variety of intracellular changes such as, decreasing cAMP leading to decreased Ca[2+] levels. The SH-SY5Y cell line study investigated whether the 5-HT[1A] agonist 8-OH-DPAT, inhibited forskolin stimulated Ca[2+] release in the SHSY-5Y cell line and whether the 5-HT[1A] antagonist p-MPPI reversed this effect using flow cytometry. The post-mortem study showed that the G-1019 allele had significantly higher 5-HT[1A] expression compared to the C allele in control hippocampal tissue. Radioligand binding data demonstrated that control samples with a GG or G/C genotype had a significantly higher 5-HT[1A] receptor density compared to samples with a CC genotype. SH-SY5Y cells differentiated with RA for 5 days or NGF and aphidicolin for 10 days had significantly increased 5-HT 1A receptor mRNA levels compared to undifferentiated cells. Western blots and immunocytochemistry confirmed the presence of the 5-HT[1A] receptor in this cell line. An increase in NUDR expression was observed at the same time there is an increase in 5-HT[1A] receptor expression in SH-SY5Y cells treated with RA or NGF and aphidicolin. Flow cytometry showed that 8-OH-DPAT efficiently diminished forskolin-stimulated increase in intracellular Ca[2+] in RA differentiated cells. 5-HT also a 5-HT[1A] agonist had a similar effect. SH-SY5Y cells treated with both p-MPPI and 8-OH-DPAT demonstrated that cells treated with p-MPPI at higher concentrations significantly increased forskolin-stimulated intracellular Ca[2+] levels and therefore effectively reversed the agonistic effect of 8-OH-DPAT. The findings presented in the post-mortem study are novel and the SH-SY5Y cell line study demonstrates that this cell line when differentiated with either RA or NGF and aphidicolin is a useful cell-line model system for studying the 5-HT[1A] receptor."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["5-HT1A receptor expression : Studies in postmortem tissue and characterisation of a model system."]}]}],"canonical_facts":{"dc:contributor.advisor":["Dalton, Caroline","Hall, Adrian","Reynolds, Gavin"],"dc:creator":["Bunn, Lindsey J."],"dc:date":["2008"],"dc:date.issued":["2008"],"dc:description.abstract":["Serotonin (5-HT) neurotransmission is involved in the psychopharmacology of several psychiatric disorders including, depression, anxiety disorders and schizophrenia. The release of 5-HT in neurons is mediated by somatodendritic 5-HT[1A] autoreceptors. The presence of 5-HT[1A] receptor is thought to be increased in depressed patients, producing a reduction in the synthesis of 5-HT. A common single nucleotide polymorphism in the promoter region of 5HT[1A] receptor C-1019G is also associated with depression and suicide. The nuclear DEAF-1 related (NUDR) protein represses the 5-HT[1A] promoter region hence regulating both 5-HT[1A] transcription and receptor expression. The project involved undertaking a post-mortem study to determine any association between the 5-HT[1A] promoter polymorphism and the expression of 5-HT[1A] receptor mRNA and receptor density in control human hippocampal brain tissue. This was achieved by genotyping human brain tissue for the 5-HT[1A] receptor polymorphism C-1019G and 5-HT[1A] receptor mRNA levels were quantified using real-time PCR. Radioligand binding was used to determine B[max] and Kd quantifying 5-HT[1A] receptor density. The SHSY-5Y neuroblastoma cell line is a well characterised cell line model used in neurotransmitter studies when differentiated. The 5-HT[1A] receptor couples to Gi proteins inhibiting AC activity and hence mediating a variety of intracellular changes such as, decreasing cAMP leading to decreased Ca[2+] levels. The SH-SY5Y cell line study investigated whether the 5-HT[1A] agonist 8-OH-DPAT, inhibited forskolin stimulated Ca[2+] release in the SHSY-5Y cell line and whether the 5-HT[1A] antagonist p-MPPI reversed this effect using flow cytometry. The post-mortem study showed that the G-1019 allele had significantly higher 5-HT[1A] expression compared to the C allele in control hippocampal tissue. Radioligand binding data demonstrated that control samples with a GG or G/C genotype had a significantly higher 5-HT[1A] receptor density compared to samples with a CC genotype. SH-SY5Y cells differentiated with RA for 5 days or NGF and aphidicolin for 10 days had significantly increased 5-HT 1A receptor mRNA levels compared to undifferentiated cells. Western blots and immunocytochemistry confirmed the presence of the 5-HT[1A] receptor in this cell line. An increase in NUDR expression was observed at the same time there is an increase in 5-HT[1A] receptor expression in SH-SY5Y cells treated with RA or NGF and aphidicolin. Flow cytometry showed that 8-OH-DPAT efficiently diminished forskolin-stimulated increase in intracellular Ca[2+] in RA differentiated cells. 5-HT also a 5-HT[1A] agonist had a similar effect. SH-SY5Y cells treated with both p-MPPI and 8-OH-DPAT demonstrated that cells treated with p-MPPI at higher concentrations significantly increased forskolin-stimulated intracellular Ca[2+] levels and therefore effectively reversed the agonistic effect of 8-OH-DPAT. The findings presented in the post-mortem study are novel and the SH-SY5Y cell line study demonstrates that this cell line when differentiated with either RA or NGF and aphidicolin is a useful cell-line model system for studying the 5-HT[1A] receptor."],"dc:format":["application/pdf"],"dc:identifier.uri":["https://shura.shu.ac.uk/19409/1/10694290.pdf"],"dc:language":["en"],"dc:publisher.commercial":["Sheffield Hallam University,"],"dc:publisher.department":["Department Not Provided."],"dc:publisher.institution":["Sheffield Hallam University (United Kingdom)."],"dc:relation.isreferencedby":["https://shura.shu.ac.uk/19409/"],"dc:title":["5-HT1A receptor expression : Studies in postmortem tissue and characterisation of a model system."],"dc:type":["Thesis"],"dc:type.qualificationlevel":["doctoral"],"dc:type.qualificationname":["phd"]},"updated_at":"2026-07-24T06:31:14Z"}