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Characterization of pro-opiomelanocortin gene variants and their effect on carcass traits in beef cattleDeobald, Heather Maureen 17 September 2009
Pro-opiomelanocortin is a prohormone that codes for many different peptides, some of which are involved in the appetite pathway. A single nucleotide polymorphism c.288C>T in pro-opiomelanocortin (POMC) was previously demonstrated to be associated with hot carcass weight (HCW) and shipping weight (Ship wt) in cattle. While developing a commercial real time PCR test for the POMC c.288C>T we identified a 12 bp deletion (POMC c.293_304delTTGGGGGCGCGG). The deletion results in the removal of four amino acids; valine, two glycine, and alanine. The deletion does not cause a frame shift. Both the POMC c.288C>T SNP and the deletion were genotyped in 386 crossbred steers, and evaluated for associations with carcass traits. The animals with one copy of the deletion had a significantly lower end-of-background rib-eye area (P=0.04) and carcass rib-eye area (P=0.03) when compared to animals without the deletion. A significant association with the POMC c.288C>T SNP was found with start of finishing weight (SOF WT); (P=0.04), HCW (P=0.02), average fat and grade fat (P=0.05), carcass rib-eye area (REA); (P=0.03) and marbling (P=0.02). These results suggest that it would be beneficial for beef producers to know both the deletion and the POMC c.288C>T SNP genotypes when making marketing and culling decisions.
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Characterization of pro-opiomelanocortin gene variants and their effect on carcass traits in beef cattleDeobald, Heather Maureen 17 September 2009 (has links)
Pro-opiomelanocortin is a prohormone that codes for many different peptides, some of which are involved in the appetite pathway. A single nucleotide polymorphism c.288C>T in pro-opiomelanocortin (POMC) was previously demonstrated to be associated with hot carcass weight (HCW) and shipping weight (Ship wt) in cattle. While developing a commercial real time PCR test for the POMC c.288C>T we identified a 12 bp deletion (POMC c.293_304delTTGGGGGCGCGG). The deletion results in the removal of four amino acids; valine, two glycine, and alanine. The deletion does not cause a frame shift. Both the POMC c.288C>T SNP and the deletion were genotyped in 386 crossbred steers, and evaluated for associations with carcass traits. The animals with one copy of the deletion had a significantly lower end-of-background rib-eye area (P=0.04) and carcass rib-eye area (P=0.03) when compared to animals without the deletion. A significant association with the POMC c.288C>T SNP was found with start of finishing weight (SOF WT); (P=0.04), HCW (P=0.02), average fat and grade fat (P=0.05), carcass rib-eye area (REA); (P=0.03) and marbling (P=0.02). These results suggest that it would be beneficial for beef producers to know both the deletion and the POMC c.288C>T SNP genotypes when making marketing and culling decisions.
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Hepatic Dysfunctions in C57/BL6 mice after Liver-based POMC OverexpressionLu, Chuan-hsiu 04 February 2010 (has links)
The pro-opiomelanocortin (POMC) prohormone produces several biologically active peptides, including £\-melanocyte-stimulating hormones (£\-MSH, £]-MSH, £^-MSH), corticotrophin (ACTH) and £]-endorphin. POMC-expressing neurons in the brain play a major role in the control of pain, energy homeostasis, pigmentation, adrenocortical function, and sebaceous gland lipid production. Recently, the peripheral POMC system is under active investigation to delineate their pathogenic roles in metabolic diseases such as Cushing¡¦s syndrome and obesity. In the present study, we employed adenovirus gene delivery system to achieve POMC overexpression in the livers of adult C57/BL6 mice. In the endocrine system of adrenal glands, hepatic POMC overexpression mice display hypertrophy the ACTH levels elevated concentrations in the blood, the ACTH receptor, melanocortin type 2 receptor (MC2-R) were decrease. This phenomenon explained the local adrenal gland tissue was inhibiting and feedback from central hypothalamic-pituitary- adrenal axis. Meanwhile, we investigated the islets of Langerhans in hepatic POMC overexpression mice, the insulin were disappear but the glucagon were constant, these reflect the blood sugar were loss of balance, maybe progress to metabolic syndrome. Subsequently, hepatic POMC overexpression resulted in liver injuries that the ALT and AST levels were significantly higher, the fat accumulation in the liver and the glycogen were diminished to nearly 1/4 of basal levels. Evidence the hepatic POMC overexpression induced inflammatory and fatty changes in the livers of mice. In summary, POMC gene delivery induces systemic POMC overexpression and results in fatty liver and adrenal dysfunction, which may facilitates a mice model for Cushing¡¦s-like metabolic syndrome.
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Lateral Parabrachial Choline Acetyltransferase Neurons and the Decision to EatTatera, Walter James 13 June 2023 (has links)
Food choice is a modifiable health factor which involves neural, hormonal, and metabolic signals. The lateral parabrachial nucleus is a brain structure in the pons that integrates multiple aspects of food choice. It receives information from the homeostatic melanocortin hypothalamic system and projects to the mesolimbic dopamine reward system. The lateral parabrachial is molecularly diverse and expresses the acetylcholine synthesis enzyme: choline acetyltransferase (ChAT). In this study, we used ChAT-CRE mice to investigate the anatomical projections, the calcium dynamics, and the causal role of the LPBN ChAT neurons. Anatomical projection results were produced using CRE dependent viral vectors to express mRuby tagged synaptophysin, the highest output being the central amygdala. Calcium dynamics were measured at the amygdala using the genetically encoded calcium indicator GCaMP. The dynamics around the decision to consume food were seen to be different between fasted and sated satiety states. Activation of the circuit showed a pronounced latency to food consumption and time to finish for a single calorie of food. These data demonstrate a possible node that integrates homeostatic feeding information and relays it to higher order brain systems that modify reward value. / Master of Science / Health can be impacted by the food an individual decides to eat, and this choice is controlled by the brain. There are many regions of the brain that are recruited when an individual decides to eat, but the two major circuits recruited are the homeostatic feeding circuit and the reward feeding circuit. The homeostatic feeding circuit involves the hypothalamus, the structure that controls basic essential functions of the body and circulating hunger hormones to signal energy availability. The second circuit is the reward circuitry which uses the neurotransmitter dopamine to signal pleasure and motivation for food. At the middle of the two circuits sits the parabrachial nucleus which expresses choline acetyltransferase, the enzyme that creates the neurotransmitter acetylcholine. To harness the molecular and anatomical specificity, we employed viral dependent protein expression to measure the anatomical output, the activity when a mouse is engaged in feeding behavior, and the causal role of the identified circuit during feeding behavior. The results showed the anatomical output to be the central amygdala, a modifier of food reward and value. The activity of the cells while feeding was seen to be higher when sated, and the activation of the circuit saw an increased latency to eat food and increased the time to consume a calorie. Together, we have demonstrated a circuit from the parabrachial nucleus the amygdala which integrates homeostatic information and projects to a brain structure that modifies food value and reward.
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Caracterização e efeitos do ACTH nas células progenitoras do córtex adrenal durante sua regeneração em animais UbiquitinaC-Cre/ERT2 Pomc Flox/Flox. / Characterization and effect of ACTH in progenitor cells of the adrenal cortex during regeneration in UbiquitinC-Cre/ERT2 POMC Flox / Flox animals.Costa, Ismael Cabral 27 September 2016 (has links)
Existem evidências na literatura que demonstram a existência de células indiferenciadas na capsula adrenal, e que o ACTH poderia estimular estas células. Porém não se sabe quais os genes e vias que desencadeiam esta resposta. Através de animais Cre-Lox induzível por Tamoxifeno, silenciamos o gene Pomc em camundongos adultos e avaliamos o efeito do ACTH nessas células. Foram utilizadas placas de PCR array para análise de genes relacionados com células progenitoras em amostras obtidas pela técnica de rolamento, e validação por PCRq com amostras microdissecadas da zona capsular/subcapsular da adrenal. Após caracterização dos animais com o gene Pomc silenciado e tratamentos com ACTH observamos o aumento da expressão de genes relacionados com as vias Wnt, Igf1 e Notch. Esses dados corroboram evidencias descritas na literatura que mostram a importância dessas vias no desenvolvimento e manutenção do córtex adrenal, e sugerem o envolvimento do ACTH nesses processos que envolvem as células progenitoras do córtex adrenal. / There is evidence in the literature demonstrating the existence of stem cells in the adrenal capsule, and that ACTH could stimulate these cells. However, it remains unknown which genes and pathways that trigger this response. By using a tamoxifen-inducible Cre-Lox mice strain, we knocked-out Pomc gene in adult mice and evaluated the effect of ACTH in these cells. PCR array technique was used to determine the expression level of key genes related to progenitor cells in samples obtained by the technique of \"rolling bearing\". Also, we validated the data by qPCR using samples from microdissected capsular areas of the adrenal gland. After characterization of animal model, the results show that treatment with ACTH increase the expression of genes related to Wnt, Igf1 and Notch pathways. These data corroborate with the literature, reinforcing the importance of these pathways in the development and maintenance of the adrenal cortex, and also suggesting the involvement of ACTH in these processes involving the progenitor cells of the adrenal cortex.
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Caracterização e efeitos do ACTH nas células progenitoras do córtex adrenal durante sua regeneração em animais UbiquitinaC-Cre/ERT2 Pomc Flox/Flox. / Characterization and effect of ACTH in progenitor cells of the adrenal cortex during regeneration in UbiquitinC-Cre/ERT2 POMC Flox / Flox animals.Ismael Cabral Costa 27 September 2016 (has links)
Existem evidências na literatura que demonstram a existência de células indiferenciadas na capsula adrenal, e que o ACTH poderia estimular estas células. Porém não se sabe quais os genes e vias que desencadeiam esta resposta. Através de animais Cre-Lox induzível por Tamoxifeno, silenciamos o gene Pomc em camundongos adultos e avaliamos o efeito do ACTH nessas células. Foram utilizadas placas de PCR array para análise de genes relacionados com células progenitoras em amostras obtidas pela técnica de rolamento, e validação por PCRq com amostras microdissecadas da zona capsular/subcapsular da adrenal. Após caracterização dos animais com o gene Pomc silenciado e tratamentos com ACTH observamos o aumento da expressão de genes relacionados com as vias Wnt, Igf1 e Notch. Esses dados corroboram evidencias descritas na literatura que mostram a importância dessas vias no desenvolvimento e manutenção do córtex adrenal, e sugerem o envolvimento do ACTH nesses processos que envolvem as células progenitoras do córtex adrenal. / There is evidence in the literature demonstrating the existence of stem cells in the adrenal capsule, and that ACTH could stimulate these cells. However, it remains unknown which genes and pathways that trigger this response. By using a tamoxifen-inducible Cre-Lox mice strain, we knocked-out Pomc gene in adult mice and evaluated the effect of ACTH in these cells. PCR array technique was used to determine the expression level of key genes related to progenitor cells in samples obtained by the technique of \"rolling bearing\". Also, we validated the data by qPCR using samples from microdissected capsular areas of the adrenal gland. After characterization of animal model, the results show that treatment with ACTH increase the expression of genes related to Wnt, Igf1 and Notch pathways. These data corroborate with the literature, reinforcing the importance of these pathways in the development and maintenance of the adrenal cortex, and also suggesting the involvement of ACTH in these processes involving the progenitor cells of the adrenal cortex.
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Hårfollikelns struktur, funktion och hårpigmenteringens genetiska reglering hos däggdjuren : Samt hur detta kan användas som modell i gymnasieksolan för att ge en djupare förståelse för genetiska interaktioner / The Function and Structure of the Hair Follicle, and the Genetical Regulation of Hair Pigmentation in Mammals : And how it can be Used as a Model in Upper Secondary School to Attain a Deeper Understanding of Genetical InteractionsSöderlund, Leo January 2020 (has links)
Hårfollikeln är en struktur som hittas hos alla däggdjur. Hår skyddar kroppen från UV-ljus, medverkar i kroppens värmereglering och har flera kommunikativa funktioner. Hårets varierande färg inom och mellan arter är både en fascinerande och intresseväckande egenskap som länge har studerats som en modell för genetisk nedärvning. I denna litteraturstudie ges en genomgång av hårfollikelns struktur och funktion, genetiken bakom hårets pigmentering samt didaktiska utmaningar i genetikundervisningen. Interaktioner mellan generna MC1R, ASIP (agouti) och POMC förklaras och exemplifieras. Dessutom diskuteras hur fårfollikeln och de pigmentreglerande generna kan användas i gymnasieskolan som ett exempel för komplicerade genetiska interaktioner. / The hair follicle is a structure found in all mammals. Hair protects the body from UV-induced damage, assists the body in its thermoregulation and has several communicative functions. The great variation in hair colour, both within and between species, is a captivating and intriguing trait that has been used as a model for genetic inheritance for a long time. This literature review features the structure and function of the hair follicle, the genetics behind the pigmentation of the hair as well as didactic challenges in teaching genetics. Interactions between the genes MC1R, ASIP (agouti) and POMC is both explained and illustrated. This review also discusses how the hair follicle and the genes regulating pigmentation can be used as an example of intricate genetic interactions in the upper secondary school. / <p>På grund av Covid-19 skedde presentation, opponering och respondering skriftligt på distans.</p>
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Du mécanisme moléculaire de la résistance hormonale à la tumorigénèse dans la maladie de CushingBilodeau, Steve January 2007 (has links)
Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.
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The Therapeutic Potential and Mechanism of POMC stress Hormone for Metastatic CancerTsai, Han-en 23 August 2012 (has links)
Despite the development of novel target therapy drugs in recent years, metastatic cancer remains refractory to current cancer therapies and accounts for the majority of cancer mortalities worldwide. Metastasis consists of multiple steps including angiogenesis, extravasion, escape from immune surveillance, adhesion, and clonal expansion in different organs that a systemic therapy is required for effective control of metastasis. The pro-inflammatory nuclear factor kappa B (NF£eB) pathway plays an important role during each of these metastatic events and constitutes an excellent target for metastasis control. Stress hormone pro-opiomelanocortin (POMC) and its derived neuropeptides including corticotrophin (ACTH), £\-, £]-, and £^-melanocyte¡Vstimulating hormone (£\-, £]-, and £^-MSH), £]-endorphin are potent inhibitors of NF£eB pathway. Other than the central regulation of stress response and energy homeostasis, POMC also regulates the skin pigmentation, inflammatory processes, and immune reactions in the peripheral system. Since adenovirus¡Vmediated POMC gene delivery leads to hepatic POMC expression, it seems plausible that POMC gene therapy may elicit systemic production of anti-inflammatory POMC-derived peptides and hold promises for control of primary and metastatic cancers. In B16-F10 melanoma models, POMC gene delivery elevated the circulating ACTH levels for more than 8 weeks and suppressed the growth of established melanoma, thereby prolonging the life span of tumor-bearing mice. Moreover, combination of POMC therapy with cisplatin further enhances the survival outcome. Subsequent analysis reveals that POMC gene therapy inhibits the growth and metastasis of melanoma through apoptosis, angiogenesis inhibition, and modulation of epithelial-mesenchymal transition. Besides, £\-MSH/melanortin-1 receptor (MC-1R) pathway is involved in the POMC-mediated melanoma suppression.
To investigate whether POMC therapy could be applied to other types of tumor, we evaluated the therapeutic efficacy of POMC gene therapy in Lewis lung carcinoma (LLC) cells which lack MC-1R. Interestingly, POMC gene delivery effectively inhibited the proliferation and colony formation of LLC cells in vitro and the growth of established LLC in mice. Histological analysis indicated that POMC gene delivery attenuated LLC through proliferation inhibition, apoptosis induction, and angiogenesis blockade. Moreover, POMC gene delivery perturbed £]-catenin signaling by reducing protein levels of £]-catenin and its downstream proto-oncogenes, including cyclin D1 and c-myc. These results support the existence of an MC-1R-independent pathway for POMC gene therapy and expand the therapeutic spectrum of POMC therapy for multiple types of cancer.
To elucidate the role of host immunity in anti-neoplastic mechanism underlying POMC therapy, we compared the treatment efficacy of POMC gene therapy for B16-F10 melanoma between severe combined immune-deficient (SCID) and immune-competent C57BL/6 mice, and found similar extent of tumor suppression in both strains of mice. In addition, POMC gene therapy reduced the spleen weight and the number of circulating lymphocytes in B6 mice. These findings suggest that POMC therapy was not dependent on host immunity, yet instead induced immune suppression of animals through ACTH/cortisol production. To minimize such side effect of POMC therapy, we generated a series of adenovirus vectors encoding POMC with mutations in ACTH domain (ACTH-K15A/R17A), which fails to stimulate cortisol synthesis in vitro and in vivo. Gene delivery of ACTH (K15A/R17A) remained capable of suppressing the primary and metastatic melanoma, but had no effect on immune functions in mice. In conclusion, we have characterized the anti-neoplastic function and mechanism of POMC therapy for cancer. Furthermore, we have developed improved POMC gene vectors to minimize its adverse effect for future cancer therapy.
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Gene Delivery of POMC for treatment of Intractable PainChuang, Ming-Ju 31 July 2003 (has links)
The use of gene-based techniques to produce antinociceptive molecules has been actively investigated for treatment of neuropathic pain and trauma of central nervous system. Among the endogenous opioids, b-endorphin (b-EP) is the most potent one, which is derived from pro-opiomelanocortin (POMC). In addition to b-endorphin, POMC is also the precursor of many neuropeptides such as adrenocorticotropin hormone (ACTH), melanocyte-stimulating hormone (a-MSH), ¡Ketc. Appropriate administration of POMC gene is essential for the success of its clinical application. Thus, gene transfer approach seems to be suitable for continuous supply of b-endorphin to alleviate intractable pain. Recombinant adenovirus was used as gene delivery system for POMC because of its high titer, wide host range, and transduction efficiency. In the present study, we have generated and characterized the recombinant adenovirus encoding POMC (Ad-POMC) by PCR and western blot analysis, and detect the presence of opioid peptides including ACTH, a-MSH and b-EP by RIA and chemilluminiscent assay. GH3 cells infected with Ad-POMC showed significantly higher levels of ACTH, b-endorphin, and a¡VMSH comparing with cells of control groups. By using Ad-GFP, the optimal MOI for adenovirus vector to infect neuronal GH3 cells, glial C6 cells, hepatoma Hep3B cells, smooth muscle G8 cells, fibroblast CCD-965K cells, and endothelial EA.hy926 cells was determined at 50, 500, 50, 500, 500, and 200, respectively. The results of determining the efficiency of POMC processing in different types of cells after in vitro cell cultures gene delivery indicated that peripheral cells, though at a lower extent, are capable of cleaving POMC and releasing opioid peptides after POMC gene delivery like neuronal cells of central nervous system. In formalin test, the intrathecal POMC gene delivery significantly decreased the magnitude of the formalin-evoked flinching response phase 1 (P < 0.05) and phase 2 (P < 0.001) when compared with rats receiving saline or Ad-GFP. In conclusion, the intrathecal POMC gene delivery can produce effectively attenuation on the inflammatory pain response. So far, there have been various gene delivery studies confirming the potential role of POMC in antinociception. In the future, more experiments will be needed to characterize the effects of POMC expression on cellular lipid metabolism. This will enable us to evaluate the therapeutic potential of POMC on treatment of obesity.
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