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Lumbriculus variegatus: A novel organism for in vivo pharmacology research / CAITLIN BELLAMY

Swansea University Author: CAITLIN BELLAMY

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Abstract

For years animal models in science have been invaluable and highly beneficial to the advancement of medicine and pharmacology. Many in vivo models are protected by the Animals (Scientific Procedures) Act 1986, and there is framework put in place to replace, reduce and refine the number of animals us...

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Published: Swansea, Wales, UK 2023
Institution: Swansea University
Degree level: Master of Research
Degree name: MSc by Research
Supervisor: Seeley, Aidan., Davies, Nia. and Wallace, Melisa.
URI: https://cronfa.swan.ac.uk/Record/cronfa65177
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This means there is a call for an in vivo model that will give us insight into specific pharmacological processes, while reducing the need for vertebrate animal models in research. Here we present the fresh freshwater invertebrate, Lumbriculus variegatus, as a novel in vivo model for pharmacology research.Here we have developed two assays to measure the behavioural effects of drugs on L. variegatus when exposed to specific compounds: the stereotypical movement assay, which measures the worms stereotyped behaviours in response to stimuli, and the free locomotion assay, which measures L. variegatus unstimulated movement. We report the effects of compounds with diverse pharmacodynamic properties on L. variegatus using these assays, these include ion channel blockers, neurotransmitters and their antagonists, and drugs of abuse. Alongside this, we have also developed techniques to extract and quantify protein and DNA from this organism.Our results show that ion channel blockers, lidocaine and quinine, reduced both stimulated and unstimulated movement in L. variegatus. Stereotypical movement and free locomotion were both significantly affected when L. variegatus were exposed to ≥20 mM of dopamine and ≥50 μM of dopamine antagonist haloperidol. However, dose-dependent effects were only observed for stimulated movement when exposed to GABA, and changes were observed only at the highest concentration of 500 mM when exposed to glycine. Both stimulated and unstimulated movement was reduced when L. variegatus was exposed to ≥250 mM. L. variegatus also displayed a dose-dependent response to DNP and were unable to recover after 24 hours at 50 μM. These toxic effects were reversed by 10 and 25 μM of haloperidol, and 25 μM of sulpiride. We successfully extracted and quantified both protein and DNA from this organism.We recognise that the experiments we have conducted on L. variegatus throughout this project may not replicate the complexity of higher animals, and experiments utilising invertebrates will not fully replace studies in vertebrate species. L. variegatus have the potential to replace smaller invertebrate models where specialist equipment is needed to visualise them. An advantage of using L. variegatus for pharmacology is that they possess unique stereotypical behaviours that can be easily quantified without the need for specialist equipment. Alongside this, there is no call for special husbandry as with rodents and other larger models, therefore L. variegatus can be cultured in most laboratories, including research and educational institutions.</abstract><type>E-Thesis</type><journal/><volume/><journalNumber/><paginationStart/><paginationEnd/><publisher/><placeOfPublication>Swansea, Wales, UK</placeOfPublication><isbnPrint/><isbnElectronic/><issnPrint/><issnElectronic/><keywords>Animals, Laboratory, Education, Invertebrates, Models, Animals, Educational, Oligochaeta, Teaching</keywords><publishedDay>13</publishedDay><publishedMonth>10</publishedMonth><publishedYear>2023</publishedYear><publishedDate>2023-10-13</publishedDate><doi/><url/><notes/><college>COLLEGE NANME</college><CollegeCode>COLLEGE CODE</CollegeCode><institution>Swansea University</institution><supervisor>Seeley, Aidan., Davies, Nia. and Wallace, Melisa.</supervisor><degreelevel>Master of Research</degreelevel><degreename>MSc by Research</degreename><apcterm/><funders/><projectreference/><lastEdited>2023-12-01T15:37:36.2729519</lastEdited><Created>2023-12-01T15:30:20.3141712</Created><path><level id="1">Faculty of Medicine, Health and Life Sciences</level><level id="2">Swansea University Medical School - Biomedical Science</level></path><authors><author><firstname>CAITLIN</firstname><surname>BELLAMY</surname><order>1</order></author></authors><documents><document><filename>65177__29166__8f1abf137ea84c7283040e8d2ae22184.pdf</filename><originalFilename>2023_Bellamy_C.final.65177.pdf</originalFilename><uploaded>2023-12-01T15:37:06.4360914</uploaded><type>Output</type><contentLength>4821371</contentLength><contentType>application/pdf</contentType><version>E-Thesis – open access</version><cronfaStatus>true</cronfaStatus><documentNotes>Copyright: The Author, Caitlin Bellamy, 2023.</documentNotes><copyrightCorrect>true</copyrightCorrect><language>eng</language></document></documents><OutputDurs/></rfc1807>
spelling v2 65177 2023-12-01 Lumbriculus variegatus: A novel organism for in vivo pharmacology research e9321656ab2f6f9dc30f5e1bb3605ba3 CAITLIN BELLAMY CAITLIN BELLAMY true false 2023-12-01 For years animal models in science have been invaluable and highly beneficial to the advancement of medicine and pharmacology. Many in vivo models are protected by the Animals (Scientific Procedures) Act 1986, and there is framework put in place to replace, reduce and refine the number of animals used in research. This means there is a call for an in vivo model that will give us insight into specific pharmacological processes, while reducing the need for vertebrate animal models in research. Here we present the fresh freshwater invertebrate, Lumbriculus variegatus, as a novel in vivo model for pharmacology research.Here we have developed two assays to measure the behavioural effects of drugs on L. variegatus when exposed to specific compounds: the stereotypical movement assay, which measures the worms stereotyped behaviours in response to stimuli, and the free locomotion assay, which measures L. variegatus unstimulated movement. We report the effects of compounds with diverse pharmacodynamic properties on L. variegatus using these assays, these include ion channel blockers, neurotransmitters and their antagonists, and drugs of abuse. Alongside this, we have also developed techniques to extract and quantify protein and DNA from this organism.Our results show that ion channel blockers, lidocaine and quinine, reduced both stimulated and unstimulated movement in L. variegatus. Stereotypical movement and free locomotion were both significantly affected when L. variegatus were exposed to ≥20 mM of dopamine and ≥50 μM of dopamine antagonist haloperidol. However, dose-dependent effects were only observed for stimulated movement when exposed to GABA, and changes were observed only at the highest concentration of 500 mM when exposed to glycine. Both stimulated and unstimulated movement was reduced when L. variegatus was exposed to ≥250 mM. L. variegatus also displayed a dose-dependent response to DNP and were unable to recover after 24 hours at 50 μM. These toxic effects were reversed by 10 and 25 μM of haloperidol, and 25 μM of sulpiride. We successfully extracted and quantified both protein and DNA from this organism.We recognise that the experiments we have conducted on L. variegatus throughout this project may not replicate the complexity of higher animals, and experiments utilising invertebrates will not fully replace studies in vertebrate species. L. variegatus have the potential to replace smaller invertebrate models where specialist equipment is needed to visualise them. An advantage of using L. variegatus for pharmacology is that they possess unique stereotypical behaviours that can be easily quantified without the need for specialist equipment. Alongside this, there is no call for special husbandry as with rodents and other larger models, therefore L. variegatus can be cultured in most laboratories, including research and educational institutions. E-Thesis Swansea, Wales, UK Animals, Laboratory, Education, Invertebrates, Models, Animals, Educational, Oligochaeta, Teaching 13 10 2023 2023-10-13 COLLEGE NANME COLLEGE CODE Swansea University Seeley, Aidan., Davies, Nia. and Wallace, Melisa. Master of Research MSc by Research 2023-12-01T15:37:36.2729519 2023-12-01T15:30:20.3141712 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Biomedical Science CAITLIN BELLAMY 1 65177__29166__8f1abf137ea84c7283040e8d2ae22184.pdf 2023_Bellamy_C.final.65177.pdf 2023-12-01T15:37:06.4360914 Output 4821371 application/pdf E-Thesis – open access true Copyright: The Author, Caitlin Bellamy, 2023. true eng
title Lumbriculus variegatus: A novel organism for in vivo pharmacology research
spellingShingle Lumbriculus variegatus: A novel organism for in vivo pharmacology research
CAITLIN BELLAMY
title_short Lumbriculus variegatus: A novel organism for in vivo pharmacology research
title_full Lumbriculus variegatus: A novel organism for in vivo pharmacology research
title_fullStr Lumbriculus variegatus: A novel organism for in vivo pharmacology research
title_full_unstemmed Lumbriculus variegatus: A novel organism for in vivo pharmacology research
title_sort Lumbriculus variegatus: A novel organism for in vivo pharmacology research
author_id_str_mv e9321656ab2f6f9dc30f5e1bb3605ba3
author_id_fullname_str_mv e9321656ab2f6f9dc30f5e1bb3605ba3_***_CAITLIN BELLAMY
author CAITLIN BELLAMY
author2 CAITLIN BELLAMY
format E-Thesis
publishDate 2023
institution Swansea University
college_str Faculty of Medicine, Health and Life Sciences
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hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Biomedical Science{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Biomedical Science
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description For years animal models in science have been invaluable and highly beneficial to the advancement of medicine and pharmacology. Many in vivo models are protected by the Animals (Scientific Procedures) Act 1986, and there is framework put in place to replace, reduce and refine the number of animals used in research. This means there is a call for an in vivo model that will give us insight into specific pharmacological processes, while reducing the need for vertebrate animal models in research. Here we present the fresh freshwater invertebrate, Lumbriculus variegatus, as a novel in vivo model for pharmacology research.Here we have developed two assays to measure the behavioural effects of drugs on L. variegatus when exposed to specific compounds: the stereotypical movement assay, which measures the worms stereotyped behaviours in response to stimuli, and the free locomotion assay, which measures L. variegatus unstimulated movement. We report the effects of compounds with diverse pharmacodynamic properties on L. variegatus using these assays, these include ion channel blockers, neurotransmitters and their antagonists, and drugs of abuse. Alongside this, we have also developed techniques to extract and quantify protein and DNA from this organism.Our results show that ion channel blockers, lidocaine and quinine, reduced both stimulated and unstimulated movement in L. variegatus. Stereotypical movement and free locomotion were both significantly affected when L. variegatus were exposed to ≥20 mM of dopamine and ≥50 μM of dopamine antagonist haloperidol. However, dose-dependent effects were only observed for stimulated movement when exposed to GABA, and changes were observed only at the highest concentration of 500 mM when exposed to glycine. Both stimulated and unstimulated movement was reduced when L. variegatus was exposed to ≥250 mM. L. variegatus also displayed a dose-dependent response to DNP and were unable to recover after 24 hours at 50 μM. These toxic effects were reversed by 10 and 25 μM of haloperidol, and 25 μM of sulpiride. We successfully extracted and quantified both protein and DNA from this organism.We recognise that the experiments we have conducted on L. variegatus throughout this project may not replicate the complexity of higher animals, and experiments utilising invertebrates will not fully replace studies in vertebrate species. L. variegatus have the potential to replace smaller invertebrate models where specialist equipment is needed to visualise them. An advantage of using L. variegatus for pharmacology is that they possess unique stereotypical behaviours that can be easily quantified without the need for specialist equipment. Alongside this, there is no call for special husbandry as with rodents and other larger models, therefore L. variegatus can be cultured in most laboratories, including research and educational institutions.
published_date 2023-10-13T15:37:37Z
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score 11.012791

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