title: The Goat (Capra hircus) Mammary Gland Mitochondrial Proteome: A Study on the Effect of Weight Loss Using Blue-Native PAGE and Two-Dimensional Gel Electrophoresis.
abstract: Seasonal weight loss (SWL) is the most important limitation to animal production in the Tropical and Mediterranean regions, conditioning producer's incomes and the nutritional status of rural communities. It is of importance to produce strategies to oppose adverse effects of SWL. Breeds that have evolved in harsh climates have acquired tolerance to SWL through selection. Most of the factors determining such ability are related to changes in biochemical pathways as affected by SWL. In this study, a gel based proteomics strategy (BN: Blue-Native Page and 2DE: Two-dimensional gel electrophoresis) was used to characterize the mitochondrial proteome of the secretory tissue of the goat mammary gland. In addition, we have conducted an investigation of the effects of weight loss in two goat breeds with different levels of adaptation to nutritional stress: Majorera (tolerant) and Palmera (susceptible). The study used Majorera and Palmera dairy goats, divided in 4 sets, 2 for each breed: underfed group fed on wheat straw (restricted diet, so their body weight would be 15-20% reduced by the end of experiment), and a control group fed with an energy-balanced diet. At the end of the experimental period (22 days), mammary gland biopsies were obtained for all experimental groups. The proteomic analysis of the mitochondria enabled the resolution of a total of 277 proteins, and 148 (53%) were identified by MALDI-TOF/TOF mass spectrometry. Some of the proteins were identified as subunits of the glutamate dehydrogenase complex and the respiratory complexes I, II, IV, V from mitochondria, as well as numerous other proteins with functions in: metabolism, development, localization, cellular organization and biogenesis, biological regulation, response to stimulus, among others, that were mapped in both BN and 2DE gels. The comparative proteomics analysis enabled the identification of several proteins: NADH-ubiquinone oxidoreductase 75 kDa subunit and lamin B1 mitochondrial (up-regulated in the Palmera breed), Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2 (up-regulated in the Majorera breed) and cytochrome b-c1 complex subunit 1, mitochondrial and Chain D, Bovine F1-C8 Sub-Complex Of Atp Synthase (down-regulated in the Majorera breed) as a consequence of weight loss.
authors.affiliation: CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal.|Animal Science Department, Universidad de Las Palmas de Gran Canaria, Arucas, Gran Canaria, Spain.
authors.forename: Graziano
authors.initials: G
authors.lastname: Cugno
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnologica, Oeiras, Portugal.|ITQB/UNL - Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: José R
authors.initials: JR
authors.lastname: Parreira
authors.affiliation: CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal.|Department of Veterinary Medical Sciences, University of Bologna, Bologna, Italy.
authors.forename: Enea
authors.initials: E
authors.lastname: Ferlizza
authors.affiliation: Animal Science Department, Universidad de Las Palmas de Gran Canaria, Arucas, Gran Canaria, Spain.|Veterinary Physiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
authors.forename: Lorenzo E
authors.initials: LE
authors.lastname: Hernández-Castellano
authors.affiliation: CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal.
authors.forename: Mariana
authors.initials: M
authors.lastname: Carneiro
authors.affiliation: LIST - Luxemburg Institute of Science and Technology, Belvaux, Luxemburg.
authors.forename: Jenny
authors.initials: J
authors.lastname: Renaut
authors.affiliation: Animal Science Department, Universidad de Las Palmas de Gran Canaria, Arucas, Gran Canaria, Spain.
authors.forename: Noemí
authors.initials: N
authors.lastname: Castro
authors.affiliation: Animal Science Department, Universidad de Las Palmas de Gran Canaria, Arucas, Gran Canaria, Spain.
authors.forename: Anastasio
authors.initials: A
authors.lastname: Arguello
authors.affiliation: Instituto Canario de Investigaciones Agrarias, Valle Guerra, Tenerife, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Capote
authors.affiliation: CIIMAR, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Porto, Portugal.
authors.forename: Alexandre M O
authors.initials: AM
authors.lastname: Campos
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnologica, Oeiras, Portugal.|ITQB/UNL - Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.|CIISA-Centro Interdisciplinar de Investigação em Sanidade Animal, Lisboa, Portugal.
authors.forename: André M
authors.initials: AM
authors.lastname: Almeida
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D045222:Electron Transport Chain Complex Proteins / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006041:Goats; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D015431:Weight Loss
chemical_list: D045222:Electron Transport Chain Complex Proteins; D024101:Mitochondrial Proteins; D020543:Proteome