{"id":{"repo_id":"creighton","oai_identifier":"oai:cdr.creighton.edu:10504/4544"},"canonical_url":"https://search.dev.ndltd.org/etd/creighton/oai:cdr.creighton.edu:10504/4544","repository":{"repo_id":"creighton","name":"Creighton University","base_url":"https://cdr.creighton.edu/server/oai/request"},"display":{"title":"Regulation of ERK1/2 and SAPK/JNK Phosphorylation by Histamine","abstract":"Mitogen activated protein kinases (MAPKs) are specific serine/threonine kinases which respond to various stimuli and control various cellular activities including gene expression, mitosis, cell differentiation, and cell survival/apoptosis. Histamine is implicated in allergic disease and asthma and SAPK/JNK and ERK1/2 are involved in certain aspects of allergic inflammation such as TH2 differentiation and proliferation and apoptosis. This study was designed to investigate the effects of histamine on ERK1/2 and SAPK/JNK phosphorylation in splenocytes.|C57BL/6 splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M). Histamine at higher concentration (10<sup>-4</sup> M) increased ERK2 phosphorylation. There was, however no significant effect seen at other concentrations (10<sup>-6</sup> M - 10<sup>-11</sup> M). Surprisingly, H1 receptor agonist betahistine (10<sup>-5</sup> M), H2 agonist amthamine (10<sup>- 5</sup> M), H3 agonist methimepip (10<sup>-6</sup> M) and H4 agonist 4-methyl histamine (10<sup>-6</sup> M), all increased ERK2 phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>- 5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and H3R antagonist clobenpropit (10<sup>-5</sup> M) inhibited histamine mediated ERK2 phosphorylation suggesting that all four histamine receptor subtypes played some role in this phosphorylation. Since TNF-alpha causes phosphorylation of ERK1/2, we investigated whether histamine acted via secretion of TNF-alpha to affect ERK1/2 phosphorylation. As a consequence, TNFalpha knockout mice were used and we found that TNF-alpha was involved in ERK2 phosphorylation. There was complete inhibition of ERK2 phosphorylation by histamine via H2, H3 and H4 agonists, but effects of H1 agonist were inconclusive in TNF-alpha knockout splenocytes. This suggested that histamine indirectly affected the ERK2 phosphorylation via its effects on the secretion of TNF-alpha and H1 receptor played a role in this process.|We performed similar experiments with stress activated protein kinases/c-jun N-terminal kinases (SAPK/JNK). C57BL/6 mice splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M), phorbol 12 myristate 13-acetate (PMA) was used as a positive control and phosphorylation of SAPK/JNK was determined. Histamine inhibited phosphorylation of SAPK/JNK at high concentrations (10<sup>-4</sup> M<sup>-10-8</sup> M) and had no effect on SAPK/JNK phosphorylation at lower concentrations (10<sup>-9</sup> M - 10<sup>- 11</sup> M). Histamine receptor specific agonists were used to identify the histamine receptors involved in the inhibition of SAPK/JNK phosphorylation. H1R agonist betahistine (10-5 M) decreased the phosphorylation of SAPK/JNK. The decrease in SAPK/JNK phosphorylation by histamine was predominantly an H1 receptor effect. H2R agonist amthamine (10<sup>-5</sup> M) did not show any significant effect on SAPK/JNK phosphorylation. H3R agonist methimepip (10<sup>-6</sup> M) and H4R agonist 4-methyl histamine (10<sup>-6</sup> M), increased SAPK/JNK phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>-5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and clobenpropit (10<sup>-5</sup> M), partially reversed the histamine mediated inhibition of SAPK/JNK phosphorylation. TNF-alpha knockout mice were used to determine if histamine regulated SAPK/JNK phosphorylation via TNF-alpha. In TNF-alpha knockout mice splenocytes, histamine inhibited SAPK/JNK phosphorylation. Activation of H1 receptors inhibited SAPK/JNK phosphorylation in knockout as was the case in wild type mice. Histamine via H2 receptor inhibited SAPK/JNK phosphorylation in knockout mice, but did not show any significant effect in wild type mice. Activation of H3 receptors decreased SAPK/JNK phosphorylation in knockout mice, as opposed to an increase in wild type mice suggesting that another cytokine besides TNF-alpha was involved in SAPK/JNK phosphorylation. H4 receptor did not show any significant effect in knockout mice, but showed an increase in SAPK/JNK phosphorylation in wild type mice suggesting that TNF-alpha is required for histamine mediated effects via H4 receptors. This data showed a role of TNF-alpha in histamine-mediated effects of SAPK/JNK phosphorylation via H4 receptors.","abstract_html":"Mitogen activated protein kinases (MAPKs) are specific serine/threonine kinases which respond to various stimuli and control various cellular activities including gene expression, mitosis, cell differentiation, and cell survival/apoptosis. Histamine is implicated in allergic disease and asthma and SAPK/JNK and ERK1/2 are involved in certain aspects of allergic inflammation such as TH2 differentiation and proliferation and apoptosis. This study was designed to investigate the effects of histamine on ERK1/2 and SAPK/JNK phosphorylation in splenocytes.|C57BL/6 splenocytes were treated with different concentrations of histamine (10&lt;sup&gt;-4&lt;/sup&gt; M to 10&lt;sup&gt;-11&lt;/sup&gt; M). Histamine at higher concentration (10&lt;sup&gt;-4&lt;/sup&gt; M) increased ERK2 phosphorylation. There was, however no significant effect seen at other concentrations (10&lt;sup&gt;-6&lt;/sup&gt; M - 10&lt;sup&gt;-11&lt;/sup&gt; M). Surprisingly, H1 receptor agonist betahistine (10&lt;sup&gt;-5&lt;/sup&gt; M), H2 agonist amthamine (10&lt;sup&gt;- 5&lt;/sup&gt; M), H3 agonist methimepip (10&lt;sup&gt;-6&lt;/sup&gt; M) and H4 agonist 4-methyl histamine (10&lt;sup&gt;-6&lt;/sup&gt; M), all increased ERK2 phosphorylation. H1R antagonist pyrilamine (10&lt;sup&gt;-6&lt;/sup&gt; M), H2R antagonist ranitidine (10&lt;sup&gt;- 5&lt;/sup&gt; M), H3/H4R antagonist thioperamide (10&lt;sup&gt;-6&lt;/sup&gt; M), and H3R antagonist clobenpropit (10&lt;sup&gt;-5&lt;/sup&gt; M) inhibited histamine mediated ERK2 phosphorylation suggesting that all four histamine receptor subtypes played some role in this phosphorylation. Since TNF-alpha causes phosphorylation of ERK1/2, we investigated whether histamine acted via secretion of TNF-alpha to affect ERK1/2 phosphorylation. As a consequence, TNFalpha knockout mice were used and we found that TNF-alpha was involved in ERK2 phosphorylation. There was complete inhibition of ERK2 phosphorylation by histamine via H2, H3 and H4 agonists, but effects of H1 agonist were inconclusive in TNF-alpha knockout splenocytes. This suggested that histamine indirectly affected the ERK2 phosphorylation via its effects on the secretion of TNF-alpha and H1 receptor played a role in this process.|We performed similar experiments with stress activated protein kinases/c-jun N-terminal kinases (SAPK/JNK). C57BL/6 mice splenocytes were treated with different concentrations of histamine (10&lt;sup&gt;-4&lt;/sup&gt; M to 10&lt;sup&gt;-11&lt;/sup&gt; M), phorbol 12 myristate 13-acetate (PMA) was used as a positive control and phosphorylation of SAPK/JNK was determined. Histamine inhibited phosphorylation of SAPK/JNK at high concentrations (10&lt;sup&gt;-4&lt;/sup&gt; M&lt;sup&gt;-10-8&lt;/sup&gt; M) and had no effect on SAPK/JNK phosphorylation at lower concentrations (10&lt;sup&gt;-9&lt;/sup&gt; M - 10&lt;sup&gt;- 11&lt;/sup&gt; M). Histamine receptor specific agonists were used to identify the histamine receptors involved in the inhibition of SAPK/JNK phosphorylation. H1R agonist betahistine (10-5 M) decreased the phosphorylation of SAPK/JNK. The decrease in SAPK/JNK phosphorylation by histamine was predominantly an H1 receptor effect. H2R agonist amthamine (10&lt;sup&gt;-5&lt;/sup&gt; M) did not show any significant effect on SAPK/JNK phosphorylation. H3R agonist methimepip (10&lt;sup&gt;-6&lt;/sup&gt; M) and H4R agonist 4-methyl histamine (10&lt;sup&gt;-6&lt;/sup&gt; M), increased SAPK/JNK phosphorylation. H1R antagonist pyrilamine (10&lt;sup&gt;-6&lt;/sup&gt; M), H2R antagonist ranitidine (10&lt;sup&gt;-5&lt;/sup&gt; M), H3/H4R antagonist thioperamide (10&lt;sup&gt;-6&lt;/sup&gt; M), and clobenpropit (10&lt;sup&gt;-5&lt;/sup&gt; M), partially reversed the histamine mediated inhibition of SAPK/JNK phosphorylation. TNF-alpha knockout mice were used to determine if histamine regulated SAPK/JNK phosphorylation via TNF-alpha. In TNF-alpha knockout mice splenocytes, histamine inhibited SAPK/JNK phosphorylation. Activation of H1 receptors inhibited SAPK/JNK phosphorylation in knockout as was the case in wild type mice. Histamine via H2 receptor inhibited SAPK/JNK phosphorylation in knockout mice, but did not show any significant effect in wild type mice. Activation of H3 receptors decreased SAPK/JNK phosphorylation in knockout mice, as opposed to an increase in wild type mice suggesting that another cytokine besides TNF-alpha was involved in SAPK/JNK phosphorylation. H4 receptor did not show any significant effect in knockout mice, but showed an increase in SAPK/JNK phosphorylation in wild type mice suggesting that TNF-alpha is required for histamine mediated effects via H4 receptors. This data showed a role of TNF-alpha in histamine-mediated effects of SAPK/JNK phosphorylation via H4 receptors.","abstract_has_math":false,"creators":["Dandekar, Radhika D."],"institution":"Creighton University","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Khan, Manzoor M."],"committee_chairs":[],"committee_members":[],"year":2009,"date_issued":"2009-12","date_published":"2009-12","updated_at":"2026-07-24T01:50:02Z","subjects":[],"languages":["en_US"],"rights":["Copyright"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10504/4544","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Khan, Manzoor M."]},{"key":"dc:creator","label":"Author","values":["Dandekar, Radhika D."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2010-01-22T22:20:01Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2010-01-22T22:20:01Z"]},{"key":"dc:date.issued","label":"Date","values":["2009-12"]},{"key":"dc:publisher","label":"Institution","values":["Creighton University"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["Dandekar-thesis.pdf"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10504/4544"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Mitogen activated protein kinases (MAPKs) are specific serine/threonine kinases which respond to various stimuli and control various cellular activities including gene expression, mitosis, cell differentiation, and cell survival/apoptosis. Histamine is implicated in allergic disease and asthma and SAPK/JNK and ERK1/2 are involved in certain aspects of allergic inflammation such as TH2 differentiation and proliferation and apoptosis. This study was designed to investigate the effects of histamine on ERK1/2 and SAPK/JNK phosphorylation in splenocytes.|C57BL/6 splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M). Histamine at higher concentration (10<sup>-4</sup> M) increased ERK2 phosphorylation. There was, however no significant effect seen at other concentrations (10<sup>-6</sup> M - 10<sup>-11</sup> M). Surprisingly, H1 receptor agonist betahistine (10<sup>-5</sup> M), H2 agonist amthamine (10<sup>- 5</sup> M), H3 agonist methimepip (10<sup>-6</sup> M) and H4 agonist 4-methyl histamine (10<sup>-6</sup> M), all increased ERK2 phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>- 5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and H3R antagonist clobenpropit (10<sup>-5</sup> M) inhibited histamine mediated ERK2 phosphorylation suggesting that all four histamine receptor subtypes played some role in this phosphorylation. Since TNF-alpha causes phosphorylation of ERK1/2, we investigated whether histamine acted via secretion of TNF-alpha to affect ERK1/2 phosphorylation. As a consequence, TNFalpha knockout mice were used and we found that TNF-alpha was involved in ERK2 phosphorylation. There was complete inhibition of ERK2 phosphorylation by histamine via H2, H3 and H4 agonists, but effects of H1 agonist were inconclusive in TNF-alpha knockout splenocytes. This suggested that histamine indirectly affected the ERK2 phosphorylation via its effects on the secretion of TNF-alpha and H1 receptor played a role in this process.|We performed similar experiments with stress activated protein kinases/c-jun N-terminal kinases (SAPK/JNK). C57BL/6 mice splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M), phorbol 12 myristate 13-acetate (PMA) was used as a positive control and phosphorylation of SAPK/JNK was determined. Histamine inhibited phosphorylation of SAPK/JNK at high concentrations (10<sup>-4</sup> M<sup>-10-8</sup> M) and had no effect on SAPK/JNK phosphorylation at lower concentrations (10<sup>-9</sup> M - 10<sup>- 11</sup> M). Histamine receptor specific agonists were used to identify the histamine receptors involved in the inhibition of SAPK/JNK phosphorylation. H1R agonist betahistine (10-5 M) decreased the phosphorylation of SAPK/JNK. The decrease in SAPK/JNK phosphorylation by histamine was predominantly an H1 receptor effect. H2R agonist amthamine (10<sup>-5</sup> M) did not show any significant effect on SAPK/JNK phosphorylation. H3R agonist methimepip (10<sup>-6</sup> M) and H4R agonist 4-methyl histamine (10<sup>-6</sup> M), increased SAPK/JNK phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>-5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and clobenpropit (10<sup>-5</sup> M), partially reversed the histamine mediated inhibition of SAPK/JNK phosphorylation. TNF-alpha knockout mice were used to determine if histamine regulated SAPK/JNK phosphorylation via TNF-alpha. In TNF-alpha knockout mice splenocytes, histamine inhibited SAPK/JNK phosphorylation. Activation of H1 receptors inhibited SAPK/JNK phosphorylation in knockout as was the case in wild type mice. Histamine via H2 receptor inhibited SAPK/JNK phosphorylation in knockout mice, but did not show any significant effect in wild type mice. Activation of H3 receptors decreased SAPK/JNK phosphorylation in knockout mice, as opposed to an increase in wild type mice suggesting that another cytokine besides TNF-alpha was involved in SAPK/JNK phosphorylation. H4 receptor did not show any significant effect in knockout mice, but showed an increase in SAPK/JNK phosphorylation in wild type mice suggesting that TNF-alpha is required for histamine mediated effects via H4 receptors. This data showed a role of TNF-alpha in histamine-mediated effects of SAPK/JNK phosphorylation via H4 receptors."]},{"key":"dc:title","label":"Title","values":["Regulation of ERK1/2 and SAPK/JNK Phosphorylation by Histamine"]}]}],"canonical_facts":{"dc:contributor.advisor":["Khan, Manzoor M."],"dc:creator":["Dandekar, Radhika D."],"dc:date.accessioned":["2010-01-22T22:20:01Z"],"dc:date.available":["2010-01-22T22:20:01Z"],"dc:date.issued":["2009-12"],"dc:description.abstract":["Mitogen activated protein kinases (MAPKs) are specific serine/threonine kinases which respond to various stimuli and control various cellular activities including gene expression, mitosis, cell differentiation, and cell survival/apoptosis. Histamine is implicated in allergic disease and asthma and SAPK/JNK and ERK1/2 are involved in certain aspects of allergic inflammation such as TH2 differentiation and proliferation and apoptosis. This study was designed to investigate the effects of histamine on ERK1/2 and SAPK/JNK phosphorylation in splenocytes.|C57BL/6 splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M). Histamine at higher concentration (10<sup>-4</sup> M) increased ERK2 phosphorylation. There was, however no significant effect seen at other concentrations (10<sup>-6</sup> M - 10<sup>-11</sup> M). Surprisingly, H1 receptor agonist betahistine (10<sup>-5</sup> M), H2 agonist amthamine (10<sup>- 5</sup> M), H3 agonist methimepip (10<sup>-6</sup> M) and H4 agonist 4-methyl histamine (10<sup>-6</sup> M), all increased ERK2 phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>- 5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and H3R antagonist clobenpropit (10<sup>-5</sup> M) inhibited histamine mediated ERK2 phosphorylation suggesting that all four histamine receptor subtypes played some role in this phosphorylation. Since TNF-alpha causes phosphorylation of ERK1/2, we investigated whether histamine acted via secretion of TNF-alpha to affect ERK1/2 phosphorylation. As a consequence, TNFalpha knockout mice were used and we found that TNF-alpha was involved in ERK2 phosphorylation. There was complete inhibition of ERK2 phosphorylation by histamine via H2, H3 and H4 agonists, but effects of H1 agonist were inconclusive in TNF-alpha knockout splenocytes. This suggested that histamine indirectly affected the ERK2 phosphorylation via its effects on the secretion of TNF-alpha and H1 receptor played a role in this process.|We performed similar experiments with stress activated protein kinases/c-jun N-terminal kinases (SAPK/JNK). C57BL/6 mice splenocytes were treated with different concentrations of histamine (10<sup>-4</sup> M to 10<sup>-11</sup> M), phorbol 12 myristate 13-acetate (PMA) was used as a positive control and phosphorylation of SAPK/JNK was determined. Histamine inhibited phosphorylation of SAPK/JNK at high concentrations (10<sup>-4</sup> M<sup>-10-8</sup> M) and had no effect on SAPK/JNK phosphorylation at lower concentrations (10<sup>-9</sup> M - 10<sup>- 11</sup> M). Histamine receptor specific agonists were used to identify the histamine receptors involved in the inhibition of SAPK/JNK phosphorylation. H1R agonist betahistine (10-5 M) decreased the phosphorylation of SAPK/JNK. The decrease in SAPK/JNK phosphorylation by histamine was predominantly an H1 receptor effect. H2R agonist amthamine (10<sup>-5</sup> M) did not show any significant effect on SAPK/JNK phosphorylation. H3R agonist methimepip (10<sup>-6</sup> M) and H4R agonist 4-methyl histamine (10<sup>-6</sup> M), increased SAPK/JNK phosphorylation. H1R antagonist pyrilamine (10<sup>-6</sup> M), H2R antagonist ranitidine (10<sup>-5</sup> M), H3/H4R antagonist thioperamide (10<sup>-6</sup> M), and clobenpropit (10<sup>-5</sup> M), partially reversed the histamine mediated inhibition of SAPK/JNK phosphorylation. TNF-alpha knockout mice were used to determine if histamine regulated SAPK/JNK phosphorylation via TNF-alpha. In TNF-alpha knockout mice splenocytes, histamine inhibited SAPK/JNK phosphorylation. Activation of H1 receptors inhibited SAPK/JNK phosphorylation in knockout as was the case in wild type mice. Histamine via H2 receptor inhibited SAPK/JNK phosphorylation in knockout mice, but did not show any significant effect in wild type mice. Activation of H3 receptors decreased SAPK/JNK phosphorylation in knockout mice, as opposed to an increase in wild type mice suggesting that another cytokine besides TNF-alpha was involved in SAPK/JNK phosphorylation. H4 receptor did not show any significant effect in knockout mice, but showed an increase in SAPK/JNK phosphorylation in wild type mice suggesting that TNF-alpha is required for histamine mediated effects via H4 receptors. This data showed a role of TNF-alpha in histamine-mediated effects of SAPK/JNK phosphorylation via H4 receptors."],"dc:identifier.other":["Dandekar-thesis.pdf"],"dc:identifier.uri":["http://hdl.handle.net/10504/4544"],"dc:language.iso":["en_US"],"dc:publisher":["Creighton University"],"dc:rights":["Copyright"],"dc:title":["Regulation of ERK1/2 and SAPK/JNK Phosphorylation by Histamine"],"dc:type":["Thesis"]},"updated_at":"2026-07-24T01:50:02Z"}