Expression of key substrate cycle enzymes in rat spermatogenic cells: Fructose 1,6 bisphosphatase and 6 phosphofructose 1‐kinase

Yáñez, Alejandro J.; Bustamante, Ximena; Bertinat, Romina; Werner, E; Rauch, M.; Concha, Ilona I.; Reyes, Juan G.; Slebe, Juan C.

doi:10.1002/jcp.21077

Cited by 12 publications

(6 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This could reflect energy substrate availability, because germ cells in the luminal compartment of seminiferous tubules are believed to rely mainly on the breakdown of lactate and pyruvate provided by Sertoli cells (38,39). Independently of OXPHOS, pyruvate import into mitochondria may serve the anabolic needs of spermiogenesis, for instance for gluconeogenesis that has been suggested to occur in round spermatids in rats (40) or as a source of Acetyl-CoA for histone acetylation (41, 42), a necessary step for the massive compaction of chromatin that occurs during spermiogenesis (43). Development of reliable in vitro models of spermatogenesis will be important in the future to allow the meticulous examination of metabolic fluxes and their role in the various spermatogenic cell types.…”

Section: Discussionmentioning

confidence: 99%

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

Selective transport of pyruvate across the inner mitochondrial membrane by the mitochondrial pyruvate carrier (MPC) is a fundamental step that couples cytosolic and mitochondrial metabolism. The recent molecular identification of the MPC complex has revealed two interacting subunits, MPC1 and MPC2. Although in yeast, an additional subunit, MPC3, can functionally replace MPC2, no alternative MPC subunits have been described in higher eukaryotes. Here, we report for the first time the existence of a novel MPC subunit termed MPC1-like (MPC1L), which is present uniquely in placental mammals. MPC1L shares high sequence, structural, and topological homology with MPC1. In addition, we provide several lines of evidence to show that MPC1L is functionally equivalent to MPC1: 1) when co-expressed with MPC2, it rescues pyruvate import in a MPC-deleted yeast strain; 2) in mammalian cells, it can associate with MPC2 to form a functional carrier as assessed by bioluminescence resonance energy transfer; 3) in MPC1 depleted mouse embryonic fibroblasts, MPC1L rescues the loss of pyruvate-driven respiration and stabilizes MPC2 expression; and 4) MPC1-and MPC1L-mediated pyruvate imports show similar efficiency. However, we show that MPC1L has a highly specific expression pattern and is localized almost exclusively in testis and more specifically in postmeiotic spermatids and sperm cells. This is in marked contrast to MPC1/MPC2, which are ubiquitously expressed throughout the organism. To date, the biological importance of this alternative MPC complex during spermatogenesis in placental mammals remains unknown. Nevertheless, these findings open up new avenues for investigating the structure-function relationship within the MPC complex.The metabolic balance between glycolysis and oxidative phosphorylation (OXPHOS) 2 is of crucial importance in determining cell function and fate. Rapidly proliferating cells rely preferentially on glycolysis despite its lower yield of ATP, because glycolytic intermediates efficiently meet the anabolic needs for sustained cell growth. In rapidly dividing cells, this occurs even in the presence of oxygen, a process called aerobic glycolysis, or the Warburg effect (1, 2). Differentiated cells rely more heavily on OXPHOS to meet the energy demands associated with their specialization. In addition, a shift from glycolytic to oxidative metabolism is necessary for the differentiation process to occur (2-4), and conversely, a hallmark of cancer lies in the rewiring of cellular metabolism toward increased glycolytic flux (5). Pyruvate, the end product of glycolysis, is imported into the mitochondrial matrix where it fuels the TCA cycle and thereby provides electrons and reducing equivalents to the respiratory chain. Pyruvate-derived acetyl-CoA can also be used as an anabolic substrate for the synthesis of lipids and amino acids. Thus, elucidating the molecular mechanisms that determine the intracellular fate of pyruvate is important for understanding the control of cell metabolism and ultimately of cell funct...

show abstract

Section: Discussionmentioning

confidence: 99%

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Vanderperre¹,

Cermakova²,

Escoffier

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Cell homogenates were prepared with lyses buffer (20 mM HEPES pH 7.5, 10 mM EGTA, 2.5 mM MgCl 2 ) plus 1% NP‐40 and protease inhibitors (Calbiochem, Darmstadt, Germany). Twenty micrograms of total proteins were fractionated in 4–12% SDS–PAGE, transferred to PVDF membranes and probed overnight with primary antibodies [Yáñez et al, 2007]. Following incubation with an HRP‐conjugated secondary antibody, reaction was developed using the Pierce ECL Western Blotting Substrate (Pierce Biotechnology, Rockford, IL).…”

Section: Methodsmentioning

confidence: 99%

Altered expression and localization of insulin receptor in proximal tubule cells from human and rat diabetic kidney

Gatica¹,

Bertinat²,

Silva³

et al. 2013

J of Cellular Biochemistry

Self Cite

View full text Add to dashboard Cite

Diabetes is the major cause of end stage renal disease, and tubular alterations are now considered to participate in the development and progression of diabetic nephropathy (DN). Here, we report for the first time that expression of the insulin receptor (IR) in human kidney is altered during diabetes. We detected a strong expression in proximal and distal tubules from human renal cortex, and a significant reduction in type 2 diabetic patients. Moreover, isolated proximal tubules from type 1 diabetic rat kidney showed a similar response, supporting its use as an excellent model for in vitro study of human DN. IR protein down-regulation was paralleled in proximal and distal tubules from diabetic rats, but prominent in proximal tubules from diabetic patients. A target of renal insulin signaling, the gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK), showed increased expression and activity, and localization in compartments near the apical membrane of proximal tubules, which was correlated with activation of the GSK3β kinase in this specific renal structure in the diabetic condition. Thus, expression of IR protein in proximal tubules from type 1 and type 2 diabetic kidney indicates that this is a common regulatory mechanism which is altered in DN, triggering enhanced gluconeogenesis regardless the etiology of the disease.

show abstract

“…The authors demonstrated the liver FBPase and aldolase B expression and that the (14C) lactate uptake was followed by an accumulation of radioactive glycogen confirming an active gluconeogenic metabolism. Our data by real time PCR analysis and histochemical studies (Yañez et al, 2007(Yañez et al, , 2003, suggest that FBPase is absent in pre-meiotic cells (spermatogonia and primary spermatocytes) and sustentocytes, but it is strongly expressed in VILLARROEL-ESPÍNDOLA, F. ; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I.…”

Section: Gluconeogenesis and Metabolic Changes During Spermatogenesismentioning

confidence: 74%

“…We have demonstrated the existence of functional gluconeogenesis in testis and sperm in different species (human, dog and rats) by highlighting the presence of key enzymes in this metabolic pathway, such as the liver isoform of fructose 1,6-bisphophatase (FBPase), phosphofructose-1 kinase (PFK) and aldolase B (Albarracin et al; Yañez et al, 2007Yañez et al, , 2003. Rat spermatids were also able to convert lactate to fructose-and glucose-6-P, indicating that both glycolytic and gluconeogenic fluxes are present in these cells (Yañez et al, 2007. We have reported that mRNA and PFK protein levels are relatively similar in spermatogenic cells, but are more strongly expressed in human sustentocytes. In contrast, expression of FBPase is stronger in round and elongating spermatids as compared to other spermatogenic cells (Yañez et al, 2003).…”

Section: Gluconeogenesis and Metabolic Changes During Spermatogenesismentioning

confidence: 99%

The Many Faces of Carbohydrate Metabolism in Male Germ Cells: From Single Molecules to Active Polymers

et al. 2018

View full text Add to dashboard Cite

VILLARROEL-ESPÍNDOLA, F.; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I. The many faces of carbohydrate metabolism in male germ cells: from single molecules to active polymers. Int. J. Med. Surg. Sci., 2(4):603-619, 2015.SUMMARY: Spermatogenesis is a complex physiological process that involves cell proliferation, meiotic division and a final cell differentiation of post-meiotic cells into spermatozoa. During this process male germ cells also undergo a metabolic differentiation process, in which post-meiotic spermatogenic cells (spermatids) but not meiotic spermatogenic cells (spermatocytes) respond differentially to D-glucose metabolism, glucose transporters (GLUTs) distribution and utilization of non-hexose substrates, such as lactate/pyruvate or dihydroxyacetone. These differences might be explained by the requirement for a specific metabolic process to support cell differentiation or in some cases, cell viability. In addition, though glycogen is considered to be the main glucose store, in male germ cells this polymer may play a novel role in cell proliferation, acting as a new marker for apoptotic events in testicular tissue via a yet unknown mechanism. In this article, we summarize the main metabolic changes that occur during male germ differentiation, with a specific focus on metabolic sources during spermatocyte to spermatid transition. The latter considering that these cells come from the same cell linage as specialized cells, but are not isolated from their environment, describing the roles from single molecules to polymers on the viability of male germ cells. Chile) Scholarship for doctoral study (to F.V.E.) and AT24100011 (to F.V.E.); MECESUP (Programa de Mejoramiento de la Calidad y Equidad de la Educación Superior, Chile) Scholarship grant UCO0606 and AUS0704 (to F.V.E.). VILLARROEL-ESPÍNDOLA, F.; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I. The many faces of carbohydrate metabolism in male germ cells: from single molecules to active polymers. Int. J. Med. Surg. Sci., 2(4):603-619, 2015. VILLARROEL-ESPÍNDOLA, F.; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I. The many faces of carbohydrate metabolism in male germ cells: from single molecules to active polymers. Int. J. Med. Surg. Sci., 2(4):603-619, 2015. VILLARROEL-ESPÍNDOLA, F.; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I. The many faces of carbohydrate metabolism in male germ cells: from single molecules to active polymers. Int. J. Med. Surg. Sci., 2(4):603-619, 2015. VILLARROEL-ESPÍNDOLA, F.; MALDONADO, R.; MANCILLA, H.; RAMÍREZ, A.; VANDER STELT, K.; CERECEDA, K.; LÓPEZ, C.; CASTRO, M. A.; ANGULO, C.; SLEBE, J. C. & CONCHA, I. I. The many faces of carbohydrate metabolism in...

show abstract

Expression of key substrate cycle enzymes in rat spermatogenic cells: Fructose 1,6 bisphosphatase and 6 phosphofructose 1‐kinase

Cited by 12 publications

References 48 publications

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

MPC1-like Is a Placental Mammal-specific Mitochondrial Pyruvate Carrier Subunit Expressed in Postmeiotic Male Germ Cells

Altered expression and localization of insulin receptor in proximal tubule cells from human and rat diabetic kidney

The Many Faces of Carbohydrate Metabolism in Male Germ Cells: From Single Molecules to Active Polymers

Contact Info

Product

Resources

About