Leptin Causes the Early Inhibition of Glycolysis- and TCA Cycle-Related Genes in the Brain of Ob/Ob Mice to Restore Fertility

Carlos Fernandes Baptista, Samuel Marcos Ribeiro de Noronha, Maria de Nazareth Gamboa Ritto, Eduardo Henrique da Silva Freitas, Melquíades Pereira Júnior, Mauro Abi Haidar, Ismael Dale Cotrim Guerreiro da Silva, Silvana Aparecida Alves Corrêa de Noronha, Marisa Teresinha Patriarca

Departmento de Medicina Geral, Universidade Federal do Rio de Janeiro (UNIRIO), Rio de Janeiro, Brazil.
Gynecology Department, Escola Paulista de Medicina/Universidade Federal de S?o Paulo (EPM-UNIFESP), S?o Paulo, Brazil.
Surgery Department, Escola Paulista de Medicina/Universidade Federal de S?o Paulo (EPM-UNIFESP), S?o Paulo, Brazil.

DOI: 10.4236/ajmb.2014.43012                

Abstract

Introduction: Polycystic ovarian syndrome (PCOS) is undoubtedly the commonest androgen disorder in woman’s fertile period and certainly one of the most prevalent causes of anovulation. The syndrome has an estimated prevalence of 4% - 10% among women of childbearing age. Previously, our group demonstrated the effect of gonadal white adipose tissue transplantation from wild-type lean and fertile female mice to isogenic obese anovulatory ob/ob mice. These complex metabolic interrelationships between obesity and PCOS have yet to be fully understood. The aim of this study was to evaluate the effect of gonadal white adipose tissue (WAT) transplantation from the wild-type lean and fertile female mice to isogenic obese, anovulatory mice (Lep ob/Lep ob) on the expression of glycolysis- and TCA cycle-related genes and obtain a general view of the glucose metabolism in the brain of these animals. Methods: Fifteen ob/ob mice ranging from 2 to 3 months of age were divided into 3 experimental groups: control normal weight (n = 5), obese control (n = 5) and obese 7 days leptin treated (n = 5). The whole brains of the mice were processed for RNA extraction. The samples from each group were used to perform PCR assays using an array plate containing 84 primers to study the glucose metabolism-related genes. Results: The glycolysis- and TCA cycle-related genes were significantly downregulated. The most significantly affected genes were as follows: for glycolysis (fold regulation with p < 0.05):Pgm1,Bpgm,Aldob, andEno3 (119, 45, 18, and 28 times less, respectively); and for the TCA cycle (fold regulation with p < 0.05):Cs,Idh3b, andMdh2 (84, 27, and 37 times less, respectively).Conclusion: The seven-day leptin treated mice show a decrease in the glucose metabolism. These results confirm the ability of the adipose tissue-derived hormone leptin to regulate early crucial genes that are related to glycolysis mechanisms and to the TCA cycle. This hormone seems to revert early the central physiological conditions that are associated with PCOS; however, the morphological alterations can only be observed within a 45-day treatment.

Keywords

PCOS, Obesity, Leptin, Glycolysis, TCA Cycle, Gene Expression

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Edman, C.D., Aiman, E.J., Porter, J.C. and MacDonald, P.C. (1978) Identification of the Estrogen Product of Extraglandular Aromatization of Plasma Androstenedione. American Journal of Obstetrics and Gynecology, 130, 439-447.
[2]	Edman, C.D. and MacDonald, P.C. (1978) Effect of Obesity on Conversion of Plasma Androstenedione to Estrone in Ovulatory and Anovulator Young Women. American Journal of Obstetrics and Gynecology, 130, 456-461.
[3]	Ehrmann, D.A. (2005) Polycystic Ovary Syndrome. The New England Journal of Medicine, 352, 1223-1236. http://dx.doi.org/10.1056/NEJMra041536
[4]	Pasquali, R. and Gambineri, A. (2006) Polycystic Ovary Syndrome: A Multifaceted Disease from Adolescence to Adult Age. The New York Academy of Sciences, 1092, 158-174. http://dx.doi.org/10.1196/annals.1365.014
[5]	Goldzieher, J.W. and Green, J.A. (1962) The Polycystic Ovary. I. Clinical and Histologic Features. The Journal of Clinical Endocrinology and Metabolism, 22, 325-338. http://dx.doi.org/10.1210/jcem-22-3-325
[6]	Vigil, P., Contreras, P., Alvarado, J.L., Godoy, A., Salgado, A.M. and Cortes, M.E. (2007) Evidence of Subpopulations with Different Levels of Insulin Resistance in Women with Polycystic Ovary Syndrome. Human Reproduction, 22, 2974-2980. http://dx.doi.org/10.1093/humrep/dem302
[7]	Ingalls, A.M., Dickie, M.M. and Snell, G.D. (1950) Obese, a New Mutation in the House Mouse. Journal of Heredity, 41, 317-318.
[8]	Jones, N. and Harrison, G.A. (1957) Genetically Determined Obesity and Sterility in the Mouse. In: Harrison, R.G., Ed., Studies on Fertility, Blackwell Scientific, Oxford, 51-64.
[9]	Pereira Jr., M., Vidotti, D.B., Borra, R.C., Simões Mde, J., Da Silva, I.D. and Haidar, M.A. (2011) Involvement of GDF-9, Leptin, and IGF1 Receptors Associated with Adipose Tissue Transplantation on Fertility Restoration in Obese Anovulatory Mice. Gynecological Endocrinology, 27, 759-766. http://dx.doi.org/10.3109/09513590.2010.534330
[10]	Vilmann, L.S., Thisted, E., Baker, J.L. and Holm, J.C. (2012) Development of Obesity and Polycystic Ovary Syndrome in Adolescents. Hormone Research in Paediatrics, 78, 269-278. http://dx.doi.org/10.1159/000345310
[11]	Buggs, C. and Rosenfield, R. (2005) Polycystic Ovary Syndrome in Adolescence. Endocrinology and Metabolism Clinics of North America, 34, 677-705. http://dx.doi.org/10.1016/j.ecl.2005.04.005
[12]	Yildiz, B.O. and Azziz, R. (2010) Ovarian and Adipose Tissue Dysfunction in Polycystic Ovary Syndrome: Report of the 4th Special Scientific Meeting of the Androgen Excess and PCOS Society. Fertility and Sterility, 94, 690-693. http://dx.doi.org/10.1016/j.fertnstert.2009.03.058
[13]	Azziz, R., Woods, K.S., Reyna, R., Key, T.J., Knochenhauer, E.S. and Yildiz, B.O. (2004) The Prevalence and Features of the Polycystic Ovary Syndrome in an Unselected Population. The Journal of Clinical Endocrinology and Metabolism, 89, 2745-2749. http://dx.doi.org/10.1210/jc.2003-032046
[14]	Lecke, S.B., Mattei, F., Morsch, D.M. and Spritzer, P.M. (2011) Abdominal Subcutaneous Fat Gene Expression and Circulating Levels of Leptin and Adiponectin in Polycystic Ovary Syndrome. Fertility and Sterility, 95, 2044-2049. http://dx.doi.org/10.1016/j.fertnstert.2011.02.041
[15]	Glueck, C.J., Morrison, J.A., Daniels, S., Wang, P. and Stroop, D. (2011) Sex Hormone-Binding Globulin, Oligomenorrhea, Polycystic Ovary Syndrome, and Childhood Insulin at Age 14 Years Predict Metabolic Syndrome and Class III Obesity at Age 24 Years. The Journal of Pediatrics, 159, 308-313.
[16]	Nair, M.K., Pappachan, P., Balakrishnan, S., Leena, M.L., George, B. and Russell, P.S. (2012) Menstrual Irregularity and Poly Cystic Ovarian Syndrome among Adolescent Girls—A 2 Year Follow-Up Study. The Indian Journal of Pediatrics, 79, S69-S73.
[17]	de Zegher, F., Lopez-Bermejo, A. and Ibáñez, L. (2009) Adipose Tissue Expandability and the Early Origins of PCOS. Trends in Endocrinology Metaboilism, 20, 418-423. http://dx.doi.org/10.1016/j.tem.2009.06.003
[18]	Hickey, M., Doherty, D.A., Atkinson, H., Sloboda, D.M., Franks, S., Norman, R.J., et al. (2011) Clinical, Ultrasound and Biochemical Features of Polycystic Ovary Syndrome in Adolescents: Implications for Diagnosis. Human Reproduction, 26, 1469-1477. http://dx.doi.org/10.1016/j.tem.2009.06.003
[19]	Reinehr, T., de Sousa, G., Roth, C.L. and Andler, W. (2005) Androgens before and after Weight Loss in Obese Children. The Journal of Clinical Endocrinology & Metabolism, 90, 5588-5595. http://dx.doi.org/10.1210/jc.2005-0438
[20]	Zhang, Y., Proenca, R., Maffei, M., Barone, M., Leopold, L. and Friedman, J.M. (1994) Positional Cloning of the Mouse Obese Gene and Its Human Homologue. Nature, 372, 425-432. http://dx.doi.org/10.1038/372425a0
[21]	Mantzoros, C.S. (2001) The Role of Leptin and Hypothalamic Neuropeptides in Energy Homeostasis: Update on Leptin in Obesity. Growth Hormone and IGF Research, 11, S85-S89. http://dx.doi.org/10.1016/S1096-6374(01)80014-9
[22]	Ceddia, R.B., Koistinen, H.A., Zierath, J.R. and Sweeney, G. (2002) Analysis of Paradoxical Observations on the Association between Leptin and Insulin Resistance. The FASEB Journal, 16, 1163-1176. http://dx.doi.org/10.1096/fj.02-0158rev
[23]	Moschos, S., Chan, J.L. and Mantzoros, C.S. (2002) Leptin and Reproduction: A Review. Fertility and Sterility, 77, 433-444. http://dx.doi.org/10.1016/S0015-0282(01)03010-2
[24]	Coleman, D.L. (1978) Obese and Diabetes: Two Mutant Genes Causing Diabetes-Obesity Syndromes in Mice. Diabetologia, 14, 141-148. http://dx.doi.org/10.1007/BF00429772
[25]	Hummel, K.P., Dickie, M.M. and Coleman, D.L. (1966) Diabetes, a New Mutation in the Mouse. Science, 153, 1127-1128. http://dx.doi.org/10.1126/science.153.3740.1127
[26]	Kashima, K., Yahata, T., Fujita, K. and Tanaka, K. (2013) Polycystic Ovary Syndrome: Association of a C/T Single Nucleotide Polymorphism at Tyrosine Kinase Domain of Insulin Receptor Gene with Pathogenesis among Lean Japanese Women. Journal of Reproductive Medicine, 58, 491-496.
[27]	Burcelin, R., Thorens, B., Glauser, M., Gaillard, R.C. and Pralong, F.P. (2003) FP Gonadotropin-Releasing Hormone Secretion from Hypothalamic Neurons: Stimulation by Insulin and Potentiation by Leptin. Endocrinology, 144, 4484-4491. http://dx.doi.org/10.1210/en.2003-0457
[28]	Lake, J.K., Power, C. and Cole, T.J. (1997) Women’s Reproductive Health: The Role of Body Mass Index in Early and Adult Life. International Journal of Obesity and Related Metabolic Disorders, 21, 432-438. http://dx.doi.org/10.1038/sj.ijo.0800424
[29]	Gavrilova, O., Marcus-Samuels, B., Graham, D., Kim, J.K., Shulman, G.I., Castle, A.L., Vinson, C., Eckhaus, M., Reitman, M.L. (2000) Surgical Implantation of Adipose Tissue Reverses Diabetes in Lipoatrophic Mice. Journal of Clinical Investigation, 105, 271-278.
[30]	Khan, A., Zong-Chao, L., Efendic, S. and Landau, B.R. (1998) Glucose-6-Phosphatase Activity in the Hypothalamus of ob/ob Mice. Metabolism, Clinical and Experimental, 47, 627-629.
[31]	Hartz, A.J., Barboriak, P.N, Wong, A., Katayama, K.P. and Rimm, A.A. (1979) The Association of Obesity with Infertility and Related Menstural Abnormalities in Women. International Journal of Obesity, 3, 57-73.
[32]	Brady, L.J., Silverstein, L.J., Hoppel, C.L. and Brady, P.S. (1985) Hepatic Mitochondrial Inner Membrane Properties and Carnitine Palmitoyltransferase A and B. Effect of Diabetes and Starvation. Biochemical Journal, 232, 445-450.
[33]	Delgado, T.C., Violante, I.R., Nieto-Charques, L. and Cerdan, S. (2011) Neuroglial Metabolic Compartmentation Underlying Leptin Deficiency in the Obese ob/ob Mice as Detected by Magnetic Resonance Imaging and Spectroscopy Methods. Journal of Cerebral Blood Flow & Metabolism, 31, 2257-2266. http://dx.doi.org/10.1038/jcbfm.2011.134