Follicular metabolic alterations are associated with obesity in mares and can be mitigated by dietary supplementation.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
30 Mar 2024
30 Mar 2024
Historique:
received:
16
01
2024
accepted:
27
03
2024
medline:
31
3
2024
pubmed:
31
3
2024
entrez:
30
3
2024
Statut:
epublish
Résumé
Obesity is a growing concern in human and equine populations, predisposing to metabolic pathologies and reproductive disturbances. Cellular lipid accumulation and mitochondrial dysfunction play an important role in the pathologic consequences of obesity, which may be mitigated by dietary interventions targeting these processes. We hypothesized that obesity in the mare promotes follicular lipid accumulation and altered mitochondrial function of oocytes and granulosa cells, potentially contributing to impaired fertility in this population. We also predicted that these effects could be mitigated by dietary supplementation with a combination of targeted nutrients to improve follicular cell metabolism. Twenty mares were grouped as: Normal Weight [NW, n = 6, body condition score (BCS) 5.7 ± 0.3], Obese (OB, n = 7, BCS 7.7 ± 0.2), and Obese Diet Supplemented (OBD, n = 7, BCS 7.7 ± 0.2), and fed specific feed regimens for ≥ 6 weeks before sampling. Granulosa cells, follicular fluid, and cumulus-oocyte complexes were collected from follicles ≥ 35 mm during estrus and after induction of maturation. Obesity promoted several mitochondrial metabolic disturbances in granulosa cells, reduced L-carnitine availability in the follicle, promoted lipid accumulation in cumulus cells and oocytes, and increased basal oocyte metabolism. Diet supplementation of a complex nutrient mixture mitigated most of the metabolic changes in the follicles of obese mares, resulting in parameters similar to NW mares. In conclusion, obesity disturbs the equine ovarian follicle by promoting lipid accumulation and altering mitochondrial function. These effects may be partially mitigated with targeted nutritional intervention, thereby potentially improving fertility outcomes in the obese female.
Identifiants
pubmed: 38555310
doi: 10.1038/s41598-024-58323-0
pii: 10.1038/s41598-024-58323-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7571Subventions
Organisme : USDA National Institute of Food and Agriculture Animal Health and Disease
ID : Grant No. COLV 2021-09 / Project Accession No. 1026913
Informations de copyright
© 2024. The Author(s).
Références
Silvestris, E., de Pergola, G., Rosania, R. & Loverro, G. Obesity as disruptor of the female fertility. Reprod. Biol. Endocrinol. 16, 22 (2018).
pubmed: 29523133
pmcid: 5845358
doi: 10.1186/s12958-018-0336-z
Shah, D. K., Missmer, S. A., Berry, K. F., Racowsky, C. & Ginsburg, E. S. Effect of obesity on oocyte and embryo quality in women undergoing in vitro fertilization. Obstet. Gynecol. 118, 63–70 (2011).
pubmed: 21691164
doi: 10.1097/AOG.0b013e31821fd360
Andreas, E., Winstanley, Y. E. & Robker, R. L. Effect of obesity on the ovarian follicular environment and developmental competence of the oocyte. Curr. Opin. Endocr. Metab. Res. 18, 152–158 (2021).
doi: 10.1016/j.coemr.2021.03.013
Richani, D., Dunning, K. R., Thompson, J. G. & Gilchrist, R. B. Metabolic co-dependence of the oocyte and cumulus cells: Essential role in determining oocyte developmental competence. Hum. Reprod. Update 27, 27–47 (2021).
pubmed: 33020823
doi: 10.1093/humupd/dmaa043
Van Hoeck, V. et al. Oocyte developmental failure in response to elevated nonesterified fatty acid concentrations: Mechanistic insights. Reproduction 145, 33–44 (2013).
pubmed: 23108110
doi: 10.1530/REP-12-0174
Turner, N. & Robker, R. L. Developmental programming of obesity and insulin resistance: Does mitochondrial dysfunction in oocytes play a role?. MHR Basic Sci. Reprod. Med. 21, 23–30 (2015).
doi: 10.1093/molehr/gau042
Pratt-Phillips, S. E., Owens, K. M., Dowler, L. E. & Cloninger, M. T. Assessment of resting insulin and leptin concentrations and their association with managerial and innate factors in horses. J. Equine Vet. Sci. 30, 127–133 (2010).
doi: 10.1016/j.jevs.2010.01.060
Thatcher, C. D., Pleasant, R. S., Geor, R. J. & Elvinger, F. Prevalence of overconditioning in mature horses in southwest Virginia during the summer. J. Vet. Intern. Med. 26, 1413–1418 (2012).
pubmed: 22946995
doi: 10.1111/j.1939-1676.2012.00995.x
Ragno, V. M., Zello, G. A., Klein, C. D. & Montgomery, J. B. From table to stable: A comparative review of selected aspects of human and equine metabolic syndrome. J. Equine Vet. Sci. 79, 131–138 (2019).
pubmed: 31405493
doi: 10.1016/j.jevs.2019.06.003
Harris, P. A., Bamford, N. J. & Bailey, S. R. Equine metabolic syndrome: Evolution of understanding over two decades: A personal perspective. Anim. Prod. Sci. 60, 2103 (2020).
doi: 10.1071/AN19386
Johnson, P. J., Wiedmeyer, C. E., Messer, N. T. & Ganjam, V. K. Medical implications of obesity in horses—Lessons for human obesity. J. Diabetes Sci. Technol. 3, 163–174 (2009).
pubmed: 20046661
pmcid: 2769846
doi: 10.1177/193229680900300119
Holbrook, T. C., Tipton, T. & McFarlane, D. Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses. Vet. Immunol. Immunopathol. 145, 283–289 (2012).
pubmed: 22169327
doi: 10.1016/j.vetimm.2011.11.013
Sessions, D. R., Reedy, S. E., Vick, M. M., Murphy, B. A. & Fitzgerald, B. P. Development of a model for inducing transient insulin resistance in the mare: Preliminary implications regarding the estrous cycle12. J. Anim. Sci. 82, 2321–2328 (2004).
pubmed: 15318731
doi: 10.2527/2004.8282321x
Vick, M. M. et al. Obesity is associated with altered metabolic and reproductive activity in the mare: Effects of metformin on insulin sensitivity and reproductive cyclicity. Reprod. Fertil. Dev. 18, 609 (2006).
pubmed: 16930507
doi: 10.1071/RD06016
Sessions-Bresnahan, D. R., Schauer, K. L., Heuberger, A. L. & Carnevale, E. M. Effect of obesity on the preovulatory follicle and lipid fingerprint of equine oocytes1. Biol. Reprod. https://doi.org/10.1095/biolreprod.115.130187 (2016).
doi: 10.1095/biolreprod.115.130187
pubmed: 26632608
Morley, S. A. & Murray, J.-A. Effects of body condition score on the reproductive physiology of the broodmare: A review. J. Equine Vet. Sci. 34, 842–853 (2014).
doi: 10.1016/j.jevs.2014.04.001
Robles, M. et al. Maternal obesity increases insulin resistance, low-grade inflammation and osteochondrosis lesions in foals and yearlings until 18 months of age. PLOS ONE 13, e0190309 (2018).
pubmed: 29373573
pmcid: 5786290
doi: 10.1371/journal.pone.0190309
Gastal, E. L., de Oliveira Gastal, M., Wischral, Á. & Davis, J. The equine model to study the influence of obesity and insulin resistance in human ovarian function. Acta Sci. Vet. 39(1), s57-70 (2011).
Carnevale, E. M. The mare as an animal model for reproductive aging in the women. In Animal Models and Human Reproduction (eds Shatten, E. & Constantinescu, H.) 235–242 (Wiley, 2017).
doi: 10.1002/9781118881286.ch10
Lazzari, G. Laboratory production of equine embryos. J. Equine Vet. Sci. https://doi.org/10.1016/j.jevs.2020.103097 (2020).
doi: 10.1016/j.jevs.2020.103097
pubmed: 32563445
Benammar, A. et al. The mare: A pertinent model for human assisted reproductive technologies?. Animals 11, 2304 (2021).
pubmed: 34438761
pmcid: 8388489
doi: 10.3390/ani11082304
Carnevale, E. M., Catandi, G. D. & Fresa, K. Equine aging and the oocyte: A potential model for reproductive aging in women. J. Equine Vet. Sci. 89, 103022 (2020).
pubmed: 32563447
doi: 10.1016/j.jevs.2020.103022
Catandi, G., Obeidat, Y., Chicco, A., Chen, T. & Carnevale, E. 167 Basal and maximal oxygen consumption of oocytes from young and old mares. Reprod. Fertil. Dev. 31, 208–208 (2019).
doi: 10.1071/RDv31n1Ab167
Catandi, G. et al. 98 Effects of maternal age on oxygen consumption of oocytes and in vitro-produced equine embryos. Reprod. Fertil. Dev. 32, 175–175 (2020).
doi: 10.1071/RDv32n2Ab98
Catandi, G. D. et al. Equine maternal aging affects oocyte lipid content, metabolic function and developmental potential. Reproduction 161, 399–409 (2021).
pubmed: 33539317
pmcid: 7969451
doi: 10.1530/REP-20-0494
Obeidat, Y. M. et al. Monitoring oocyte/embryo respiration using electrochemical-based oxygen sensors. Sens. Actuators B Chem. 276, 72–81 (2018).
doi: 10.1016/j.snb.2018.07.157
Obeidat, Y. M. et al. Design of a multi-sensor platform for integrating extracellular acidification rate with multi-metabolite flux measurement for small biological samples. Biosens. Bioelectron. 133, 39–47 (2019).
pubmed: 30909011
pmcid: 6660976
doi: 10.1016/j.bios.2019.02.069
Catandi, G. D. et al. Diet affects oocyte metabolism and developmental capacity in the older mare. Am. Assoc. Equine Pract. 65, 51–52 (2019).
Catandi, G. et al. Maternal diet can alter oocyte mitochondrial number and function. J. Equine Vet. Sci. 89, 103030 (2020).
doi: 10.1016/j.jevs.2020.103030
Catandi, G. D. et al. Oocyte metabolic function, lipid composition, and developmental potential are altered by diet in older mares. Reproduction 163, 183–198 (2022).
pubmed: 37379450
pmcid: 8942336
doi: 10.1530/REP-21-0351
Gonzalez, M. B., Robker, R. L. & Rose, R. D. Obesity and oocyte quality: Significant implications for ART and emerging mechanistic insights. Biol. Reprod. 106, 338–350 (2022).
pubmed: 34918035
doi: 10.1093/biolre/ioab228
Noland, R. C. et al. Carnitine insufficiency caused by aging and overnutrition compromises mitochondrial performance and metabolic control. J. Biol. Chem. 284, 22840–22852 (2009).
pubmed: 19553674
pmcid: 2755692
doi: 10.1074/jbc.M109.032888
Muoio, D. M. et al. Muscle-specific deletion of carnitine acetyltransferase compromises glucose tolerance and metabolic flexibility. Cell Metab. 15, 764–777 (2012).
pubmed: 22560225
pmcid: 3348515
doi: 10.1016/j.cmet.2012.04.005
Vincent, J. B. New evidence against chromium as an essential trace element. J. Nutr. 147, 2212–2219 (2017).
pubmed: 29021369
doi: 10.3945/jn.117.255901
Jamilian, M. et al. The influences of chromium supplementation on glycemic control, markers of cardio-metabolic risk, and oxidative stress in infertile polycystic ovary syndrome women candidate for in vitro fertilization: A randomized, double-blind, placebo-controlled trial. Biol. Trace Elem. Res. 185, 48–55 (2018).
pubmed: 29307112
doi: 10.1007/s12011-017-1236-3
Jamilian, M. et al. Effects of chromium and carnitine co-supplementation on body weight and metabolic profiles in overweight and obese women with polycystic ovary syndrome: A randomized, double-blind, placebo-controlled trial. Biol. Trace Elem. Res. 193, 334–341 (2020).
pubmed: 30977089
doi: 10.1007/s12011-019-01720-8
Seiler, S. E. et al. Obesity and lipid stress inhibit carnitine acetyltransferase activity. J. Lipid Res. 55, 635–644 (2014).
pubmed: 24395925
pmcid: 3966698
doi: 10.1194/jlr.M043448
Gervais, A., Battista, M.-C., Carranza-Mamane, B., Lavoie, H. B. & Baillargeon, J.-P. Follicular fluid concentrations of lipids and their metabolites are associated with intraovarian gonadotropin-stimulated androgen production in women undergoing in vitro fertilization. J. Clin. Endocrinol. Metab. 100, 1845–1854 (2015).
pubmed: 25695883
doi: 10.1210/jc.2014-3649
Igosheva, N. et al. Maternal diet-induced obesity alters mitochondrial activity and redox status in mouse oocytes and zygotes. PLoS ONE 5, e10074 (2010).
pubmed: 20404917
pmcid: 2852405
doi: 10.1371/journal.pone.0010074
Yang, X. et al. Exposure to lipid-rich follicular fluid is associated with endoplasmic reticulum stress and impaired oocyte maturation in cumulus-oocyte complexes. Fertil. Steril. 97, 1438–1443 (2012).
pubmed: 22440252
doi: 10.1016/j.fertnstert.2012.02.034
Boots, C. E., Boudoures, A., Zhang, W., Drury, A. & Moley, K. H. Obesity-induced oocyte mitochondrial defects are partially prevented and rescued by supplementation with co-enzyme Q10 in a mouse model. Hum. Reprod. 31, 2090–2097 (2016).
pubmed: 27432748
pmcid: 4991662
doi: 10.1093/humrep/dew181
Sutton-McDowall, M. L. et al. Nonesterified fatty acid-induced endoplasmic reticulum stress in cattle cumulus oocyte complexes alters cell metabolism and developmental competence1. Biol. Reprod. https://doi.org/10.1095/biolreprod.115.131862 (2016).
doi: 10.1095/biolreprod.115.131862
pubmed: 26658709
Carnevale, E. M. The mare model for follicular maturation and reproductive aging in the woman. Theriogenology 69, 23–30 (2008).
pubmed: 17976712
doi: 10.1016/j.theriogenology.2007.09.011
Valckx, S. D. et al. Fatty acid composition of the follicular fluid of normal weight, overweight and obese women undergoing assisted reproductive treatment: A descriptive cross-sectional study. Reprod. Biol. Endocrinol. 12, 13 (2014).
pubmed: 24498875
pmcid: 3916060
doi: 10.1186/1477-7827-12-13
Pantasri, T. et al. Distinct localisation of lipids in the ovarian follicular environment. Reprod. Fertil. Dev. 27, 593 (2015).
pubmed: 25751151
doi: 10.1071/RD14321
Gonzalez, M. B., Lane, M., Knight, E. J. & Robker, R. L. Inflammatory markers in human follicular fluid correlate with lipid levels and Body Mass Index. J. Reprod. Immunol. 130, 25–29 (2018).
pubmed: 30174020
doi: 10.1016/j.jri.2018.08.005
Valckx, S. D. M. et al. BMI-related metabolic composition of the follicular fluid of women undergoing assisted reproductive treatment and the consequences for oocyte and embryo quality. Hum. Reprod. 27, 3531–3539 (2012).
pubmed: 23019302
doi: 10.1093/humrep/des350
Mirabi, P. et al. Does different BMI influence oocyte and embryo quality by inducing fatty acid in follicular fluid?. Taiwan. J. Obstet. Gynecol. 56, 159–164 (2017).
pubmed: 28420500
doi: 10.1016/j.tjog.2016.11.005
Ribeiro, R. M. et al. Changes in metabolic and physiological biomarkers in Mangalarga Marchador horses with induced obesity. Vet. J. 270, 105627 (2021).
pubmed: 33641803
doi: 10.1016/j.tvjl.2021.105627
Wu, L.L.-Y. et al. High-fat diet causes lipotoxicity responses in cumulus-oocyte complexes and decreased fertilization rates. Endocrinology 151, 5438–5445 (2010).
pubmed: 20861227
doi: 10.1210/en.2010-0551
Lolicato, F. et al. The cumulus cell layer protects the bovine maturing oocyte against fatty acid-induced lipotoxicity1. Biol. Reprod. https://doi.org/10.1095/biolreprod.114.120634 (2015).
doi: 10.1095/biolreprod.114.120634
pubmed: 25297544
Aardema, H. et al. Bovine cumulus cells protect maturing oocytes from increased fatty acid levels by massive intracellular lipid storage. Biol. Reprod. 88, 164–164 (2013).
pubmed: 23616596
doi: 10.1095/biolreprod.112.106062
Montani, D. A. et al. The follicular microenviroment as a predictor of pregnancy: MALDI-TOF MS lipid profile in cumulus cells. J. Assist. Reprod. Genet. 29, 1289–1297 (2012).
pubmed: 22968515
pmcid: 3510365
doi: 10.1007/s10815-012-9859-y
Montani, D. A. et al. Lipid profile of cumulus cells as a predictive tool for pregnancy outcomes. Fertil. Steril. 100, S343–S344 (2013).
doi: 10.1016/j.fertnstert.2013.07.863
El-Hayek, S., Yang, Q., Abbassi, L., FitzHarris, G. & Clarke, H. J. Mammalian oocytes locally remodel follicular architecture to provide the foundation for Germline-soma communication. Curr. Biol. 28, 1124-1131.e3 (2018).
pubmed: 29576478
pmcid: 5882553
doi: 10.1016/j.cub.2018.02.039
Altermatt, J. L., Suh, T. K., Stokes, J. E. & Carnevale, E. M. Effects of age and equine follicle-stimulating hormone (eFSH) on collection and viability of equine oocytes assessed by morphology and developmental competency after intracytoplasmic sperm injection (ICSI). Reprod. Fertil. Dev. 21, 615–623 (2009).
pubmed: 19383268
doi: 10.1071/RD08210
Dunning, K. R., Russell, D. L. & Robker, R. L. Lipids and oocyte developmental competence: The role of fatty acids and β-oxidation. Reproduction 148, R15–R27 (2014).
pubmed: 24760880
doi: 10.1530/REP-13-0251
Schooneman, M. G., Vaz, F. M., Houten, S. M. & Soeters, M. R. Acylcarnitines. Diabetes 62, 1–8 (2013).
pubmed: 23258903
doi: 10.2337/db12-0466
Koves, T. R. et al. Mitochondrial overload and incomplete fatty acid oxidation contribute to skeletal muscle insulin resistance. Cell Metab. 7, 45–56 (2008).
pubmed: 18177724
doi: 10.1016/j.cmet.2007.10.013
Várnagy, Á. et al. Acylcarnitine esters profiling of serum and follicular fluid in patients undergoing in vitro fertilization. Reprod. Biol. Endocrinol. 11, 67 (2013).
pubmed: 23866102
pmcid: 3724743
doi: 10.1186/1477-7827-11-67
Ginther, O. J. et al. Comparative study of the dynamics of follicular waves in mares and women. Biol. Reprod. 71, 1195–1201 (2004).
pubmed: 15189824
doi: 10.1095/biolreprod.104.031054
Calcaterra, V. et al. Polycystic ovary syndrome in insulin-resistant adolescents with obesity: The role of nutrition therapy and food supplements as a strategy to protect fertility. Nutrients 13, 1848 (2021).
pubmed: 34071499
pmcid: 8228678
doi: 10.3390/nu13061848
Gambineri, A. et al. Female infertility: Which role for obesity?. Int. J. Obes. Suppl. 9, 65–72 (2019).
pubmed: 31391925
pmcid: 6683114
doi: 10.1038/s41367-019-0009-1
Laskowski, D. et al. Insulin during in vitro oocyte maturation has an impact on development, mitochondria, and cytoskeleton in bovine day 8 blastocysts. Theriogenology 101, 15–25 (2017).
pubmed: 28708512
doi: 10.1016/j.theriogenology.2017.06.002
Hauck, A. K. & Bernlohr, D. A. Oxidative stress and lipotoxicity. J. Lipid Res. 57, 1976–1986 (2016).
pubmed: 27009116
pmcid: 5087875
doi: 10.1194/jlr.R066597
Li, X. et al. Targeting mitochondrial reactive oxygen species as novel therapy for inflammatory diseases and cancers. J. Hematol. Oncol. 6, 19 (2013).
pubmed: 23442817
pmcid: 3599349
doi: 10.1186/1756-8722-6-19
Leroy, J. L. M. R. et al. Maternal metabolic health and fertility: We should not only care about but also for the oocyte!. Reprod. Fertil. Dev. 35, 1–18 (2022).
pubmed: 36592978
doi: 10.1071/RD22204
Cheng, M.-H. et al. Novel microsensors revealed the impact of high maternal body weight and advanced maternal aging on individual human oocyte metabolic function. 78th Sci. Congr. Am. Soc. Reprod. Med. 118, e153 (2022).
Luzzo, K. M. et al. High fat diet induced developmental defects in the mouse: Oocyte meiotic aneuploidy and fetal growth retardation/brain defects. PLoS ONE 7, e49217 (2012).
pubmed: 23152876
pmcid: 3495769
doi: 10.1371/journal.pone.0049217
Wu, L. L. et al. Mitochondrial dysfunction in oocytes of obese mothers: Transmission to offspring and reversal by pharmacological endoplasmic reticulum stress inhibitors. Development 142, 681–691 (2015).
pubmed: 25670793
doi: 10.1242/dev.114850
Marei, W. F. A. et al. Differential effects of high fat diet-induced obesity on oocyte mitochondrial functions in inbred and outbred mice. Sci. Rep. 10, 9806 (2020).
pubmed: 32555236
pmcid: 7299992
doi: 10.1038/s41598-020-66702-6
Taherkhani, S., Suzuki, K. & Ruhee, R. T. A brief overview of oxidative stress in adipose tissue with a therapeutic approach to taking antioxidant supplements. Antioxidants 10, 594 (2021).
pubmed: 33924341
pmcid: 8069597
doi: 10.3390/antiox10040594
Nilsson, M. I. et al. A multi-ingredient supplement protects against obesity and infertility in western diet-fed mice. Nutrients 15, 611 (2023).
pubmed: 36771318
pmcid: 9921271
doi: 10.3390/nu15030611
Surai, P. F. Antioxidant Action of Carnitine: Molecular Mechanisms and Practical Applications. EC Veterinary Science 2.1, 6-84 (2015).
Raviv, S. et al. Lipid droplets in granulosa cells are correlated with reduced pregnancy rates. J. Ovarian Res. 13, 4 (2020).
pubmed: 31907049
pmcid: 6945749
doi: 10.1186/s13048-019-0606-1
Su, Y.-Q., Sugiura, K. & Eppig, J. Mouse oocyte control of granulosa cell development and function: Paracrine regulation of cumulus cell metabolism. Semin. Reprod. Med. 27, 032–042 (2009).
doi: 10.1055/s-0028-1108008
Robles, M. et al. Maternal nutrition during pregnancy affects testicular and bone development, glucose metabolism and response to overnutrition in weaned horses up to two years. PLOS ONE 12, e0169295 (2017).
pubmed: 28081146
pmcid: 5231272
doi: 10.1371/journal.pone.0169295
Robles, M. et al. Placental function and structure at term is altered in broodmares fed with cereals from mid-gestation. Placenta 64, 44–52 (2018).
pubmed: 29626980
doi: 10.1016/j.placenta.2018.02.003
Leroy, J., Van Soom, A., Opsomer, G., Goovaerts, I. & Bols, P. Reduced fertility in high-yielding dairy cows: Are the Oocyte and embryo in danger? Part II mechanisms linking nutrition and reduced oocyte and embryo quality in high-yielding dairy cows*. Reprod. Domest. Anim. 43, 623–632 (2008).
pubmed: 18384498
doi: 10.1111/j.1439-0531.2007.00961.x
Rooke, J. A. et al. Dietary carbohydrates and amino acids influence oocyte quality in dairy heifers. Reprod. Fertil. Dev. 21, 419 (2009).
pubmed: 19261219
doi: 10.1071/RD08193
Skoracka, K., Ratajczak, A. E., Rychter, A. M., Dobrowolska, A. & Krela-Kaźmierczak, I. Female fertility and the nutritional approach: The most essential aspects. Adv. Nutr. 12, 2372–2386 (2021).
pubmed: 34139003
pmcid: 8634384
doi: 10.1093/advances/nmab068
Kaczmarek, K., Janicki, B. & Głowska, M. Insulin resistance in the horse: A review. J. Appl. Anim. Res. 44, 424–430 (2016).
doi: 10.1080/09712119.2015.1091340
Henneke, D. R., Potter, G. D., Kreider, J. L. & Yeates, B. F. Relationship between condition score, physical measurements and body fat percentage in mares. Equine Vet. J. 15, 371–372 (1983).
pubmed: 6641685
doi: 10.1111/j.2042-3306.1983.tb01826.x
Kane, R. A., Fisher, M., Parrett, D. & Lawrence, L. M. [Proceedings of the] 10th Equine Nutrition and Physiology Symposium, June 11–13, 1987, the Fort Collins Marriott, Colorado State University (Equine Nutrition and Physiology Society, 1987).
Carter, R. A., Geor, R. J., Burton Staniar, W., Cubitt, T. A. & Harris, P. A. Apparent adiposity assessed by standardised scoring systems and morphometric measurements in horses and ponies. Vet. J. 179, 204–210 (2009).
pubmed: 18440844
doi: 10.1016/j.tvjl.2008.02.029
Gentry, L. R. et al. The relationship between body condition score and ultrasonic fat measurements in mares of high versus low body condition. J. Equine Vet. Sci. 24, 198–203 (2004).
doi: 10.1016/j.jevs.2004.04.009
Carnevale, E. M. Advances in collection, transport and maturation of equine oocytes for assisted reproductive techniques. Vet. Clin. North Am. Equine Pract. 32, 379–399 (2016).
pubmed: 27726987
doi: 10.1016/j.cveq.2016.07.002
Larsen, S. et al. The best approach: Homogenization or manual permeabilization of human skeletal muscle fibers for respirometry?. Anal. Biochem. 446, 64–68 (2014).
pubmed: 24161612
doi: 10.1016/j.ab.2013.10.023
Li Puma, L. C. et al. Experimental oxygen concentration influences rates of mitochondrial hydrogen peroxide release from cardiac and skeletal muscle preparations. Am. J. Physiol.-Regul. Integr. Comp. Physiol. 318, R972–R980 (2020).
pubmed: 32233925
doi: 10.1152/ajpregu.00227.2019
Pfaffl, M. W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29, 45e–445 (2001).
doi: 10.1093/nar/29.9.e45
Dieckmann-Schuppert, A. & Schnittler, H.-J. A simple assay for quantification of protein in tissue sections, cell cultures, and cell homogenates, and of protein immobilized on solid surfaces. Cell Tissue Res. 288, 119–126 (1997).
pubmed: 9042779
doi: 10.1007/s004410050799
Reisz, J. A., Zheng, C., D’Alessandro, A. & Nemkov, T. Untargeted and semi-targeted lipid analysis of biological samples using mass spectrometry-based metabolomics. In High-Throughput Metabolomics: Methods and Protocols (ed. D’Alessandro, A.) 121–135 (Springer, 2019). https://doi.org/10.1007/978-1-4939-9236-2_8 .
doi: 10.1007/978-1-4939-9236-2_8
Smith, C. A., Want, E. J., O’Maille, G., Abagyan, R. & Siuzdak, G. XCMS: Processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. Anal. Chem. 78, 779–787 (2006).
pubmed: 16448051
doi: 10.1021/ac051437y
Tautenhahn, R., Böttcher, C. & Neumann, S. Highly sensitive feature detection for high resolution LC/MS. BMC Bioinform. 9, 504 (2008).
doi: 10.1186/1471-2105-9-504
Broeckling, C. D., Afsar, F. A., Neumann, S., Ben-Hur, A. & Prenni, J. E. RAMClust: A novel feature clustering method enables spectral-matching-based annotation for metabolomics data. Anal. Chem. 86, 6812–6817 (2014).
pubmed: 24927477
doi: 10.1021/ac501530d
Cheng, M.-H., Chicco, A. J., Ball, D. & Chen, T. W. Analysis of mitochondrial oxygen consumption and hydrogen peroxide release from cardiac mitochondria using electrochemical multi-sensors. Sens. Actuators B Chem. 360, 131641. https://doi.org/10.1016/j.snb.2022.131641 (2022).
doi: 10.1016/j.snb.2022.131641