Phosphorus-31 nuclear magnetic resonance studies of spermatozoa from the boar, ram, goat and bull

Robitaille, Pierre‐Marie; Robitaille, Patricia A.; Martin, P. A.; Brown, George

doi:10.1016/0305-0491(87)90141-6

Cited by 19 publications

(11 citation statements)

References 28 publications

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“…• C showed no ATP resonances, which is in line with studies by Robitaille et al (1987) and Kalic et al (1997). An oxygen supply system during NMR spectroscopy was used to demonstrate that ATP can be regenerated from AMP.…”

Section: Discussionsupporting

confidence: 91%

“…Email: kamp@mail.uni-mainz.de Unlike spermatozoa from many other vertebrates and invertebrates (Robitaille et al, 1987;Kamp et al, 1996), boar spermatozoa do not contain phosphorylcreatine and creatine kinase or any other phosphagens. However, it has been suggested that a phosphagen is necessary for the transport of energy-rich phosphate from the mitochondria in the midpiece to the distal dyneinATPases (for review, see Kaldis et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Energy metabolism and intracellular pH in boar spermatozoa

Kamp¹,

Büsselmann²,

Jones³

et al. 2003

Reproduction

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The effect of energy metabolism on intracellular pH was studied in boar spermatozoa using nuclear magnetic resonance (NMR) spectroscopy and confocal microscopy with the pH-sensitive dye seminaphthorhodafluor (SNARF-1). Freshly ejaculated spermatozoa had a high adenylate energy charge (AEC = 0.8), which decreased to 0.6 under aerobic conditions and to 0.2 under anaerobic conditions. Correspondingly, no ATP resonances but high AMP resonance were visible in 31 P-NMR-spectra of the spermatozoa. When an artificial oxygen buffer (Fluosol) and a purpose-built air supply system were used during 31 P-NMR data acquisition, ATP resonances reappeared whereas the AMP resonance disappeared. Boar spermatozoa kept under aerobic conditions have intracellular compartments that differ markedly in pH, as demonstrated by both 31 P-NMR spectroscopy and confocal microscopy. Using confocal microscopy, the midpiece of the flagellum in which all mitochondria are located was identified as an acidic compartment (pH i-mp 6.7). The intracellular pH of both the head (pH i-h ) and the long principal piece of the flagellum (pH i-pp ) were 7.2 and, thus, only slightly below the extracellular pH (pH e 7.3). Storage of spermatozoa in a glucose-free medium at 15• C when they are immotile slowly shifted the pH i-mp from 6.7 to 6.9 within 20 h, whereas pH i-h and pH i-pp remained unchanged (pH 7.1-7.2). When glucose was present in the medium, all visible compartments of the spermatozoa as well as the medium were acidified to pH 6.2 within 20 h. Under these conditions a resonance at 4.8 ppm appeared representing glycerol 3-phosphate.

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Section: Discussionsupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Energy metabolism and intracellular pH in boar spermatozoa

Kamp¹,

Büsselmann²,

Jones³

et al. 2003

Reproduction

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“…The pHi of mammalian sperm, including humans, has been evaluated using different fluorescent indicators (Florman et al, 1989 ; Vredenburgh-Wilberg and Parrish, 1995 ; Brook et al, 1996 ; Hamamah et al, 1996 ; Cross and Razy-Faulkner, 1997 ), [ 31 P]-NMR (Smith et al, 1985 ; Robitaille et al, 1987 ), and also by the distribution of a radioactive amine (Gatti et al, 1993 ; Hamamah et al, 1996 ), resulting in pHi approximately 6.7–7.2. However, there are few reports showing an increase in pHi during capacitation in human sperm.…”

Section: Extracellular and Intracellular Ph In Human Spermmentioning

confidence: 99%

Molecular Basis of Human Sperm Capacitation

Molina

Luque

Balestrini

et al. 2018

Front. Cell Dev. Biol.

176

186

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In the early 1950s, Austin and Chang independently described the changes that are required for the sperm to fertilize oocytes in vivo. These changes were originally grouped under name of “capacitation” and were the first step in the development of in vitro fertilization (IVF) in humans. Following these initial and fundamental findings, a remarkable number of observations led to characterization of the molecular steps behind this process. The discovery of certain sperm-specific molecules and the possibility to record ion currents through patch-clamp approaches helped to integrate the initial biochemical observation with the activity of ion channels. This is of particular importance in the male gamete due to the fact that sperm are transcriptionally inactive. Therefore, sperm must control all these changes that occur during their transit through the male and female reproductive tracts by complex signaling cascades that include post-translational modifications. This review is focused on the principal molecular mechanisms that govern human sperm capacitation with particular emphasis on comparing all the reported pieces of evidence with the mouse model.

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“…Vertebrate sperm contain a flagellar axoneme that structurally resembles those of eukaryotic microbes, and while sperm contains a PK, it is not flagellar‐specific. The observation that phosphagen levels vary 100‐fold between sperm of different animal species seems to conflict with PKs having a critical function in their flagella (Robitaille et al, ; Tombes and Shapiro, ). Moreover, mouse sperm function was impaired significantly by a defect in phosphagen biosynthesis, but not by a PK knockout (Steeghs et al, ; Schmidt et al, ).…”

Section: Discussionmentioning

confidence: 99%

Phosphagen kinase function in flagellated spores of the oomycete Phytophthora infestans integrates transcriptional regulation, metabolic dynamics and protein retargeting

Kagda

Ah‐Fong

et al. 2018

Molecular Microbiology

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Flagellated spores play important roles in the infection of plants and animals by many eukaryotic microbes. The oomycete Phytophthora infestans, which causes potato blight, expresses two phosphagen kinases (PKs). These enzymes store energy in taurocyamine, and are hypothesized to resolve spatial and temporal imbalances between rates of ATP creation and use in zoospores. A dimeric PK is found at low levels in vegetative mycelia, but high levels in ungerminated sporangia and zoospores. In contrast, a monomeric PK protein is at similar levels in all tissues, although is transcribed primarily in mycelia. Subcellular localization studies indicate that the monomeric PK is mitochondrial. In contrast, the dimeric PK is cytoplasmic in mycelia and sporangia but is retargeted to flagellar axonemes during zoosporogenesis. This supports a model in which PKs shuttle energy from mitochondria to and through flagella. Metabolite analysis indicates that deployment of the flagellar PK is coordinated with a large increase in taurocyamine, synthesized by sporulation-induced enzymes that were lost during the evolution of zoospore-lacking oomycetes. Thus, PK function is enabled by coordination of the transcriptional, metabolic and protein targeting machinery during the life cycle. Since plants lack PKs, the enzymes may be useful targets for inhibitors of oomycete plant pathogens.

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Phosphorus-31 nuclear magnetic resonance studies of spermatozoa from the boar, ram, goat and bull

Cited by 19 publications

References 28 publications

Energy metabolism and intracellular pH in boar spermatozoa

Energy metabolism and intracellular pH in boar spermatozoa

Molecular Basis of Human Sperm Capacitation

Phosphagen kinase function in flagellated spores of the oomycete Phytophthora infestans integrates transcriptional regulation, metabolic dynamics and protein retargeting

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