The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Tipo de documento: | Tese |
Idioma: | eng |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da USP |
Texto Completo: | http://www.teses.usp.br/teses/disponiveis/11/11139/tde-13032018-180420/ |
Resumo: | Energetic efficiency is important for health (e.g. genesis of obesity in humans), socio-economically important for meat production systems (e.g. feed cost to produce high quality protein) and important for the environment (e.g. use of natural resources and production of green house gases for meat production). Mitochondria are organelles that play an essential role in cellular metabolism and homeostasis related to energy utilization. These processes involve several proteins to ensure continuous availability of energy to the cells. The Shc proteins play a key role in substrate oxidation and energy metabolism. Additionally, the mitochondrial uncoupling proteins (UCPs) participate in physiological processes that may account for variation in energy expenditures in tissues. However, the mechanisms behind energy expenditure in animals are largely unknown. Thus, in order to study the energy metabolism and mitochondria function, studies using a nutritional, biochemical and molecular approaches were conducted with mice and cattle. The purpose of the first study was to determine if Shc proteins influence the metabolic response to acute (5-7 days) feeding of a high fat diet (HFD). To this end, whole animal energy expenditure and substrate oxidation were measured in the Shc knockout (ShcKO) and wild-type (WT) male mice consuming either a control or HFD diet. The activities of enzymes of glycolysis, the citric acid cycle, electron transport chain (ETC), and β-oxidation were investigated in liver and skeletal muscle. The study showed that ShcKO increases (P < 0.05) energy expenditure (EE) adjusted for either total body weight or lean mass. This change in EE could explain the decrease in weight gain observed in ShcKO versus WT mice fed an HFD. Thus, our results indicate that Shc proteins should be considered as potential targets for developing interventions to mitigate weight gain on HFD by stimulating EE. Although decreased levels of Shc proteins influenced the activity of some enzymes in response to high fat feeding, such as increasing the activity of acyl-CoA dehydrogenase, it did not produce concerted changes in enzymes of glycolysis, citric acid cycle or the ETC. However, the physiological significance of these changes in enzyme activities remains to be determined. The purpose of experiment 2 was to study the association among heat production, blood parameters and mitochondrial DNA (mtDNA) copy number in Nellore bulls with high and low residual feed intake (RFI). The RFI values were obtained by regression of dry mater intake (DMI) in relation to average daily gain and mid-test metabolic body weight. Thus, 18 animals (9 in each group) were individually fed in a feedlot for 98 days. The heart rate (HR) of bulls was monitored for 4 consecutive days and used to calculate the estimated heat production (EHP). Electrodes were fitted to bulls with stretch belts and oxygen consumption was obtained using a facemask connected to the gas analyzer and HR was simultaneously measured for 15 minutes period. Daily EHP was calculated multiplying oxygen pulse (O2P) by the average HR, assuming 4.89 kcal/L of O2. Blood parameters such as hematocrit, hemoglobin, and glucose were assayed between 45 and 90 days. Immediately after slaughter, liver, muscle and adipose tissues (subcutaneous and visceral fat) were collected and, subsequently, mtDNA copy number per cell was quantified in tissues by quantitative real-time PCR. The proteome of hepatic tissue and levels of mitochondrial UCPs were also investigated. We found similar EHP and O2 consumption between RFI groups, while low RFI bulls (more efficient in feed conversion) shown lower HR, hemoglobin and hematocrit percentage (P < 0.05), confirming previous data from our group. In addition, 71 protein spots in liver were differentially expressed (P < 0.05) and no differences were detected for UCPs levels between RFI groups. Finally, there was no association between amounts of mtDNA and the RFI phenotypes, suggesting that mitochondrial abundance in liver, muscle, and adipose tissue was similar between efficient and inefficient groups. However, additional studies to confirm this hypothesis are needed. |
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The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approachBioquímica da eficiência alimentar, metabolismo energético e função mitocondrial, uma abordagem animal e molecularBioenergéticaBioenergeticsBioquímica celularCell biochemistryEnergy metabolismMetabolismoMitochondrial contentMitochondrial DNAMitocôndriaEnergetic efficiency is important for health (e.g. genesis of obesity in humans), socio-economically important for meat production systems (e.g. feed cost to produce high quality protein) and important for the environment (e.g. use of natural resources and production of green house gases for meat production). Mitochondria are organelles that play an essential role in cellular metabolism and homeostasis related to energy utilization. These processes involve several proteins to ensure continuous availability of energy to the cells. The Shc proteins play a key role in substrate oxidation and energy metabolism. Additionally, the mitochondrial uncoupling proteins (UCPs) participate in physiological processes that may account for variation in energy expenditures in tissues. However, the mechanisms behind energy expenditure in animals are largely unknown. Thus, in order to study the energy metabolism and mitochondria function, studies using a nutritional, biochemical and molecular approaches were conducted with mice and cattle. The purpose of the first study was to determine if Shc proteins influence the metabolic response to acute (5-7 days) feeding of a high fat diet (HFD). To this end, whole animal energy expenditure and substrate oxidation were measured in the Shc knockout (ShcKO) and wild-type (WT) male mice consuming either a control or HFD diet. The activities of enzymes of glycolysis, the citric acid cycle, electron transport chain (ETC), and β-oxidation were investigated in liver and skeletal muscle. The study showed that ShcKO increases (P < 0.05) energy expenditure (EE) adjusted for either total body weight or lean mass. This change in EE could explain the decrease in weight gain observed in ShcKO versus WT mice fed an HFD. Thus, our results indicate that Shc proteins should be considered as potential targets for developing interventions to mitigate weight gain on HFD by stimulating EE. Although decreased levels of Shc proteins influenced the activity of some enzymes in response to high fat feeding, such as increasing the activity of acyl-CoA dehydrogenase, it did not produce concerted changes in enzymes of glycolysis, citric acid cycle or the ETC. However, the physiological significance of these changes in enzyme activities remains to be determined. The purpose of experiment 2 was to study the association among heat production, blood parameters and mitochondrial DNA (mtDNA) copy number in Nellore bulls with high and low residual feed intake (RFI). The RFI values were obtained by regression of dry mater intake (DMI) in relation to average daily gain and mid-test metabolic body weight. Thus, 18 animals (9 in each group) were individually fed in a feedlot for 98 days. The heart rate (HR) of bulls was monitored for 4 consecutive days and used to calculate the estimated heat production (EHP). Electrodes were fitted to bulls with stretch belts and oxygen consumption was obtained using a facemask connected to the gas analyzer and HR was simultaneously measured for 15 minutes period. Daily EHP was calculated multiplying oxygen pulse (O2P) by the average HR, assuming 4.89 kcal/L of O2. Blood parameters such as hematocrit, hemoglobin, and glucose were assayed between 45 and 90 days. Immediately after slaughter, liver, muscle and adipose tissues (subcutaneous and visceral fat) were collected and, subsequently, mtDNA copy number per cell was quantified in tissues by quantitative real-time PCR. The proteome of hepatic tissue and levels of mitochondrial UCPs were also investigated. We found similar EHP and O2 consumption between RFI groups, while low RFI bulls (more efficient in feed conversion) shown lower HR, hemoglobin and hematocrit percentage (P < 0.05), confirming previous data from our group. In addition, 71 protein spots in liver were differentially expressed (P < 0.05) and no differences were detected for UCPs levels between RFI groups. Finally, there was no association between amounts of mtDNA and the RFI phenotypes, suggesting that mitochondrial abundance in liver, muscle, and adipose tissue was similar between efficient and inefficient groups. However, additional studies to confirm this hypothesis are needed.A eficiência energética é importante para a saúde humana (gênese da obesidade), sistemas de produção de carne (custo dos alimentos para produzir proteínas de alta qualidade) e para o meio ambiente (uso de recursos naturais e mitigação de gases de efeito estufa). As mitocôndrias são organelas que desempenham papel central no metabolismo e homeostase relacionada a utilização da energia. Nas células, diversas proteínas são importantes para melhorar a eficiência energética. Como exemplos, as proteínas de sinalização Shc são fundamentais na oxidação de substratos e metabolismo energético e, nas mitocôndrias, existem as proteínas desacopladoras (UCPs), que participam do gasto energético e produção de calor. Entretanto, os mecanismos que controlam o gasto energético nos animais ainda é bastante desconhecido. Assim, para estudar o metabolismo energético e a função das mitocôndrias foram conduzidos dois estudos utilizando-se estratégias nutricionais, bioquímicas e moleculares com camundongos (1) e bovinos (2). Objetivou-se, no estudo 1, determinar se as proteínas Shc influenciam a resposta metabólica à alimentação contendo dieta rica em gordura (HFD) por 7 dias. Enzimas da via glicolítica, ciclo de Krebs, cadeia transportadora de elétron (CTE) e β-oxidação foram analisadas no fígado e músculo de camundongos com baixa expressão de Shc (knockout ou ShcKO) e comuns (wild-type ou WT) submetidos à uma dieta controle ou à HFD. O gasto energético foi medido por câmara calorimétrica de respiração nos animais. O genótipo ShcKO apresentou maior gasto energético (P < 0.05) ajustado para o peso corporal total ou massa magra. Essa mudança poderia explicar o menor ganho de peso observado no genótipo ShcKO comparado ao WT quando consumindo a HFD. Esses resultados sugerem que as proteínas Shc podem contribuir no desenvolvimento de estratégias para mitigar o ganho de peso. Embora a redução dos níveis de Shc (ShcKO) tenha modificado a atividade de enzimas da β-oxidação em resposta a HFD, tal condição não produziu mudanças semelhantes na via glicolítica, ciclo de Krebs ou CTE. Por isso, mais estudos são necessários para compreender a significância fisiológica dessas alterações. No experimento 2, objetivou-se estudar a associação entre produção de calor, variáveis sanguíneas e número de cópias de DNA mitocondrial (mtDNA) em bovinos Nelore agrupados pelo consumo alimentar residual (CAR). O CAR foi obtido por regressão do consumo de matéria seca em relação ao ganho de peso diário e peso metabólico do teste de desempenho (fase de crescimento). Assim, 18 bovinos (9 alto CAR versus 9 baixo CAR) foram confinados em baias individuais por 98 dias (fase de terminação). Os batimentos cardíacos (BC) dos bovinos foram monitorados por quatro dias consecutivos e, então, utilizados para o cálculo da produção de calor estimada (PCe). O consumo e pulso de oxigênio (O2) foram obtidos por meio de analisador de gás conectado à uma máscara facial, com medição simultânea dos BC por 15 minutos. A PCe diária foi calculada por multiplicação do pulso de O2 pela média dos BC, assumindo-se a constante 4.89 kcal/L de O2. Foram analisadas variáveis sanguíneas como hematócrito, hemoglobina e glicose (alto vs. baixo CAR). Imediatamente após o abate dos animais, amostras de fígado, músculo e tecido adiposo foram coletadas para determinação do mtDNA por PCR em tempo real. Adicionalmente, o proteoma do tecido hepático e os níveis de UCPs nos tecidos foram também investigados. Não houve diferença para PCe e consumo de O2 (P > 0.05) entre os grupos experimentais, entretanto, os animais baixo CAR (mais eficientes em conversão alimentar) demonstraram menor BC, concentração de hemoglobina e percentagem de hematócrito (P < 0.05), confirmando resultados previamente observados em nossos estudos. No fígado, 71 spots proteicos foram diferentes (P < 0.05) entre os grupos alto e baixo CAR, mas nenhuma diferença foi observada para os níveis de UCPs no músculo, fígado ou tecido adiposo. Por fim, não existiu diferença (P > 0.05) entre o número de cópias do mtDNA por célula entre os fenótipos estudados, sugerindo que o número de mitocôndrias e possivelmente a fosforilação oxidativa foi semelhante entre os grupos de animais eficientes e ineficientes. Contudo, são necessários estudos adicionais para confirmar essa hipótese.Biblioteca Digitais de Teses e Dissertações da USPBonilha, Sarah Figueiredo MartinsLanna, Dante Pazzanese DuarteBaldassini, Welder Angelo2017-08-11info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://www.teses.usp.br/teses/disponiveis/11/11139/tde-13032018-180420/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2018-07-19T20:50:39Zoai:teses.usp.br:tde-13032018-180420Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212018-07-19T20:50:39Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach Bioquímica da eficiência alimentar, metabolismo energético e função mitocondrial, uma abordagem animal e molecular |
title |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
spellingShingle |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach Baldassini, Welder Angelo Bioenergética Bioenergetics Bioquímica celular Cell biochemistry Energy metabolism Metabolismo Mitochondrial content Mitochondrial DNA Mitocôndria |
title_short |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
title_full |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
title_fullStr |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
title_full_unstemmed |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
title_sort |
The biochemistry of feed efficiency, energy metabolism, and mitochondrial function, an animal and molecular approach |
author |
Baldassini, Welder Angelo |
author_facet |
Baldassini, Welder Angelo |
author_role |
author |
dc.contributor.none.fl_str_mv |
Bonilha, Sarah Figueiredo Martins Lanna, Dante Pazzanese Duarte |
dc.contributor.author.fl_str_mv |
Baldassini, Welder Angelo |
dc.subject.por.fl_str_mv |
Bioenergética Bioenergetics Bioquímica celular Cell biochemistry Energy metabolism Metabolismo Mitochondrial content Mitochondrial DNA Mitocôndria |
topic |
Bioenergética Bioenergetics Bioquímica celular Cell biochemistry Energy metabolism Metabolismo Mitochondrial content Mitochondrial DNA Mitocôndria |
description |
Energetic efficiency is important for health (e.g. genesis of obesity in humans), socio-economically important for meat production systems (e.g. feed cost to produce high quality protein) and important for the environment (e.g. use of natural resources and production of green house gases for meat production). Mitochondria are organelles that play an essential role in cellular metabolism and homeostasis related to energy utilization. These processes involve several proteins to ensure continuous availability of energy to the cells. The Shc proteins play a key role in substrate oxidation and energy metabolism. Additionally, the mitochondrial uncoupling proteins (UCPs) participate in physiological processes that may account for variation in energy expenditures in tissues. However, the mechanisms behind energy expenditure in animals are largely unknown. Thus, in order to study the energy metabolism and mitochondria function, studies using a nutritional, biochemical and molecular approaches were conducted with mice and cattle. The purpose of the first study was to determine if Shc proteins influence the metabolic response to acute (5-7 days) feeding of a high fat diet (HFD). To this end, whole animal energy expenditure and substrate oxidation were measured in the Shc knockout (ShcKO) and wild-type (WT) male mice consuming either a control or HFD diet. The activities of enzymes of glycolysis, the citric acid cycle, electron transport chain (ETC), and β-oxidation were investigated in liver and skeletal muscle. The study showed that ShcKO increases (P < 0.05) energy expenditure (EE) adjusted for either total body weight or lean mass. This change in EE could explain the decrease in weight gain observed in ShcKO versus WT mice fed an HFD. Thus, our results indicate that Shc proteins should be considered as potential targets for developing interventions to mitigate weight gain on HFD by stimulating EE. Although decreased levels of Shc proteins influenced the activity of some enzymes in response to high fat feeding, such as increasing the activity of acyl-CoA dehydrogenase, it did not produce concerted changes in enzymes of glycolysis, citric acid cycle or the ETC. However, the physiological significance of these changes in enzyme activities remains to be determined. The purpose of experiment 2 was to study the association among heat production, blood parameters and mitochondrial DNA (mtDNA) copy number in Nellore bulls with high and low residual feed intake (RFI). The RFI values were obtained by regression of dry mater intake (DMI) in relation to average daily gain and mid-test metabolic body weight. Thus, 18 animals (9 in each group) were individually fed in a feedlot for 98 days. The heart rate (HR) of bulls was monitored for 4 consecutive days and used to calculate the estimated heat production (EHP). Electrodes were fitted to bulls with stretch belts and oxygen consumption was obtained using a facemask connected to the gas analyzer and HR was simultaneously measured for 15 minutes period. Daily EHP was calculated multiplying oxygen pulse (O2P) by the average HR, assuming 4.89 kcal/L of O2. Blood parameters such as hematocrit, hemoglobin, and glucose were assayed between 45 and 90 days. Immediately after slaughter, liver, muscle and adipose tissues (subcutaneous and visceral fat) were collected and, subsequently, mtDNA copy number per cell was quantified in tissues by quantitative real-time PCR. The proteome of hepatic tissue and levels of mitochondrial UCPs were also investigated. We found similar EHP and O2 consumption between RFI groups, while low RFI bulls (more efficient in feed conversion) shown lower HR, hemoglobin and hematocrit percentage (P < 0.05), confirming previous data from our group. In addition, 71 protein spots in liver were differentially expressed (P < 0.05) and no differences were detected for UCPs levels between RFI groups. Finally, there was no association between amounts of mtDNA and the RFI phenotypes, suggesting that mitochondrial abundance in liver, muscle, and adipose tissue was similar between efficient and inefficient groups. However, additional studies to confirm this hypothesis are needed. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-08-11 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.teses.usp.br/teses/disponiveis/11/11139/tde-13032018-180420/ |
url |
http://www.teses.usp.br/teses/disponiveis/11/11139/tde-13032018-180420/ |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
|
dc.rights.driver.fl_str_mv |
Liberar o conteúdo para acesso público. info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Liberar o conteúdo para acesso público. |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
|
dc.publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da USP instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Biblioteca Digital de Teses e Dissertações da USP |
collection |
Biblioteca Digital de Teses e Dissertações da USP |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br |
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1815257005403668480 |