Multiple Glycolytic Enzymes Are Tightly Bound to the Fibrous Sheath of Mouse Spermatozoa1

Krisfalusi, Michelle; Miki, Kazuo; Magyar, Patricia L.; O’Brien, Deborah A.

doi:10.1095/biolreprod.105.049684

Cited by 171 publications

(178 citation statements)

References 64 publications

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“…Mitochondria, localized in the sperm tail midpiece, have long been considered to be a major source of ATP production because of their high efficiency of energy production via oxidative respiration (19,20). Recent evidence, however, suggests that locally produced ATP via less efficient, but high throughput glycolysis in the principle piece of the sperm tail is the major energy source for sperm motility in the mouse (21)(22)(23). In our Cul4b Vasa mutants, the reduction in mitochondrial membrane potential in spermatozoa may contribute to lowered ATP production.…”

Section: Spermatozoa In Cul4b Vasa Testis Are Immotile-anatomical Examentioning

confidence: 78%

Cell Autonomous and Nonautonomous Function of CUL4B in Mouse Spermatogenesis

Yin¹,

Liu

Yang

et al. 2016

Journal of Biological Chemistry

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CUL4B ubiquitin ligase belongs to the cullin-RING ubiquitin ligase family. Although sharing many sequence and structural similarities, CUL4B plays distinct roles in spermatogenesis from its homologous protein CUL4A. We previously reported that genetic ablation of Cul4a in mice led to male infertility because of aberrant meiotic progression. In the present study, we generated Cul4b germ cell-specific conditional knock-out (Cul4b Protein degradation via the ubiquitin-proteasome system plays a critical role during mammalian spermatogenesis. Timely removal of outlived proteins is crucial to ensure progression through different phases of spermatogenesis including mitotic, meiotic divisions, and postmeiotic morphogenesis. The ubiquitin-proteasome system selects its targets via a group of E3 ubiquitin ligases, which upon interaction with E2 ubiquitin-conjugating enzymes tag substrates for proteasomal degradation by the 26S proteasome. The largest mammalian E3 ligase family is the CRL (cullin-RING finger ubiquitin ligase) family, which includes eight members. Each cullin family member serves as a molecular scaffold and forms an E3 ligase complex with substrate-recruiting receptors and a linker protein (1). The Cul4b gene, located on the X chromosome, shares extensive sequence homology and functional redundancy with the other CRL4 family member, Cul4a. Both proteins employ DDB1 (DNA damage-binding protein 1) as the linker protein, which in turn recruits common or distinct DDB1-CUL4 associated factors for substrate binding. The DDB1-CUL4-mediated protein modification/degradation is involved in critical cellular processes including DNA replication, DNA repair, cell cycle control, and histone modifications (2).Male infertility accounts for ϳ40 -50% of infertility cases in humans (3, 4). Many factors contribute to male infertility; however, the vast majority of male infertility cases are associated with oligozoospermia (decreased sperm number), asthenozoospermia (reduced sperm motility), and/or teratozoospermia (abnormal sperm morphology). In the mammalian testis, a single pluripotent spermatogonial stem cell (SSC) 2 undergoes several rounds of mitotic divisions followed by meiotic divisions and complex postmeiotic morphogenesis, eventually giving rise to a cohort of mature spermatozoa. We have previously reported that the CRL4 proteins exhibited complementary expression patterns in adult mouse testis, where CUL4A was predominantly detected in primary spermatocytes, whereas CUL4B was highly expressed in Sertoli cells, spermatogonia, and spermatids (5). The absence of CUL4B in spermatocytes is likely due to meiotic sex chromosome inactivation, a transient X chromosome inactivation caused by sex chromosome condensation and subsequent gene silencing (6). However, the

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Section: Spermatozoa In Cul4b Vasa Testis Are Immotile-anatomical Examentioning

confidence: 78%

Cell Autonomous and Nonautonomous Function of CUL4B in Mouse Spermatogenesis

Yin¹,

Liu

Yang

et al. 2016

Journal of Biological Chemistry

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“…Glycolysis is a metabolic pathway that nets two molecules of ATP and pyruvate for every single molecule of glucose that is present in virtually all organisms. Interestingly, glycolysis has been shown to be crucial for producing ATP in mammalian spermatozoa (17,18,(49)(50)(51)(52)(53)(54)(55)(56)(57); thus, decreased glycolytic enzyme levels could be a reason for asthenozoospermia in Tcte1-null mice. To verify results of the TMT quantification, three glycolytic enzymes-hexokinase-2 (HK2), pyruvate kinase (PKM), and phosphoglycerate mutase 1 (PGAM1)-were evaluated using commercially available antibodies for Western blot analysis.…”

Section: Resultsmentioning

confidence: 99%

“…Glycolysis is essential for motility because much of the ATP used by dynein is generated via the glycolytic pathway and not through the electron transport chain (17,18,(49)(50)(51)(52)(53)(54)(55)(56)(57). It is tempting to speculate that TCTE1 influences the glycolytic pathway; however, glycolysis is thought to occur at the fibrous sheath, as many glycolytic enzymes are covalently attached to that structure (40,57). The IFT machinery may link TCTE1 and the fibrous sheath.…”

Section: Discussionmentioning

confidence: 99%

TCTE1 is a conserved component of the dynein regulatory complex and is required for motility and metabolism in mouse spermatozoa

Castaneda

Hua

Miyata

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Flagella and cilia are critical cellular organelles that provide a means for cells to sense and progress through their environment. The central component of flagella and cilia is the axoneme, which comprises the "9+2" microtubule arrangement, dynein arms, radial spokes, and the nexin-dynein regulatory complex (N-DRC). Failure to properly assemble components of the axoneme leads to defective flagella and in humans leads to a collection of diseases referred to as ciliopathies. Ciliopathies can manifest as severe syndromic diseases that affect lung and kidney function, central nervous system development, bone formation, visceral organ organization, and reproduction. T-Complex-Associated-Testis-Expressed 1 (TCTE1) is an evolutionarily conserved axonemal protein present from Chlamydomonas (DRC5) to mammals that localizes to the N-DRC. Here, we show that mouse TCTE1 is testis-enriched in its expression, with its mRNA appearing in early round spermatids and protein localized to the flagellum. TCTE1 is 498 aa in length with a leucine rich repeat domain at the C terminus and is present in eukaryotes containing a flagellum. Knockout of Tcte1 results in male sterility because Tcte1-null spermatozoa show aberrant motility. Although the axoneme is structurally normal in Tcte1 mutant spermatozoa, Tcte1-null sperm demonstrate a significant decrease of ATP, which is used by dynein motors to generate the bending force of the flagellum. These data provide a link to defining the molecular intricacies required for axoneme function, sperm motility, and male fertility. male infertility | asthenozoospermia | glycolysis | mutant mouse | testis-specific gene F lagella are ancient, analogous cellular structures used for locomotion and as sensory organelles present in all three domains of life (bacteria, archaea, and eukaryotes). The advantages conferred by this organelle are highlighted by the flagella's apparent independent evolution in all three domains (1-3). Of all of the different flagella present among eukaryotes, flagella attached to gametes play a critical function in uniting gametes for fertilization and the perpetuation of a species. Mammalian spermatozoa have a specialized flagellum that contains a midpiece, principal piece, and end piece with the axoneme running along the entire length (4). The flagellum equips sperm with the capability to deliver half of the male's genetic material to the female gamete, the oocyte. In addition to flagella, eukaryotes contain another related structure called cilia. The defining feature of flagella and cilia is the axoneme, the "9+2" microtubule arraignment of two central pairs of microtubules surrounded by nine pairs of microtubule doublets (5). The microtubule motor dynein is anchored to the outer microtubules and responsible for generating the force required to produce the beating pattern of flagella and cilia (6). The force generated by dynein causes sliding of the microtubules among each other; however, the nexin complex anchors the microtubules in place. The nexin complex [or nexin-dynei...

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“…Sperm ATP concentration is thought to be high in the sperm midpiece mitochondria and sperm tail principal piece, but not necessarily in the sperm acrosome. There are at least three different sources of ATP generation in the spermatozoa [RodriguezMiranda et al 2008], including oxidative phosphorylation in the sperm mitochondria [Halangk et al 1985], glycolysis in the fibrous sheath of the sperm tail principal piece [Krisfalusi et al 2006], and the conversion of ADP to ATP by adenylate kinase [Schoff et al 1989;Foresta et al 1992]. Of particular interest to the present study is the detection of adenylate kinase-dependent KATP-potassium channels in the sperm acrosome [Acevedo et al 2006].…”

Section: Discussionmentioning

confidence: 99%

Sperm-Surface ATP in Boar Spermatozoa is Required for Fertilization: Relevance to Sperm Proteasomal Function

Park

Kim

et al. 2009

Systems Biology in Reproductive Medicine

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Extracellular ATP has been implicated in a number of cellular events, including mammalian sperm function. The complement of ATP-dependent sperm proteins includes six subunits of the 26S proteasome, a multi-subunit protease specific to ubiquitinated substrate-proteins. Proteolysis of ubiquitinated proteins by the 26S proteasome is necessary for the success of mammalian fertilization, including but not limited to acrosomal exocytosis (AE) and spermzona pellucida (ZP) penetration. The 26S proteasome is uniquely present on the sperm acrosomal surface during mammalian, ascidian, and invertebrate fertilization. The proteasome is a multi-subunit protease complex of B2 MDa composed of the 19S regulatory complex and a 20S proteolytic core. Integrity of the 19S complex is maintained by six 19S ATPase subunits (PSMC1 through PSMC6). Consequently, we hypothesized that fertilization will be blocked by the depletion of sperm-surface associated ATP (ssATP). Depletion of ssATP by the Solanum tuberosum apyrase, a 49 kDa, non-cell permeant enzyme, significantly reduced the ATP content measured by an adapted luminescence-ATP assay from which all permeabilizing agents were excluded. Addition of active apyrase to porcine in vitro fertilization (IVF) medium caused a concentration dependent reduction in the overall fertilization rate. No such outcomes were observed in control groups using heat-inactivated apyrase. Apyrase treatment altered the band pattern of 19S ATPase subunits PSMC1 (Rpt2) and PSMC4 (Rpt3) in Western blotting, suggesting that it had an effect on the integrity of the sperm proteasomal 19S complex. Apyrase only altered the proteasomal core activities slightly, since these activities are not directly dependent on external ATP. In contrast, sperm treatment with MG132, a specific inhibitor of the proteasomal core chymotrypsin-like activity, inhibited the target proteolytic activity, but also induced a compensatory elevation in proteasomal peptidylglutamyl peptide hydrolase activity. Altogether, the present data provide an important missing piece of evidence in support of the ssATP-dependent, proteasomal-proteolytic model of sperm-ZP interactions.

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Multiple Glycolytic Enzymes Are Tightly Bound to the Fibrous Sheath of Mouse Spermatozoa1

Cited by 171 publications

References 64 publications

Cell Autonomous and Nonautonomous Function of CUL4B in Mouse Spermatogenesis

Cell Autonomous and Nonautonomous Function of CUL4B in Mouse Spermatogenesis

TCTE1 is a conserved component of the dynein regulatory complex and is required for motility and metabolism in mouse spermatozoa

Sperm-Surface ATP in Boar Spermatozoa is Required for Fertilization: Relevance to Sperm Proteasomal Function

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