Adaptive expression pattern of different proteins involved in cellular calcium homeostasis in denervated rat vas deferens

Quintas, Luis Eduardo M.; Cunha, Valéria M.N.; Scaramello, Christianne Brêtas Vieira; Silva, Cláudia L.M. da; Caricati‐Neto, Afonso; Lafayette, Simone S.L.; Jurkiewicz, Aron; Noël, François

doi:10.1016/j.ejphar.2005.10.006

Cited by 12 publications

(10 citation statements)

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“…The presence in the vas deferens of molecular components responsible for Ca 2+ transport and therefore for intracellular Ca 2+ homeostasis, which is crucial for contraction, was demonstrated one decade ago [7]. Under conditions in which contraction is impaired – denervated or rapamycin-treated rat vas deferens – the sarco(endo) plasmic reticulum proteins are down-regulated [8,9], an event that could be associated with a decrease in reproductive capacity [6,10]. With respect to redox signaling, it has recently been demonstrated that reactive oxygen species (ROS) are implicated in male infertility [11].…”

Section: Introductionmentioning

confidence: 99%

Undernutrition Affects Cell Survival, Oxidative Stress, Ca2+ Handling and Signaling Pathways in Vas Deferens, Crippling Reproductive Capacity

et al. 2013

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BackgroundThe aim of this work was to investigate the mechanisms by which chronic malnutrition (CM) affects vas deferens function, leading to compromised reproductive capacity. Previous studies have shown that maternal malnutrition affects the reproductive tracts of adult male offspring. However, little is known about the effects of CM, a widespread life-long condition that persists from conception throughout growth to adult life.Methodology/Principal FindingsYoung adult male rats, which were chronically malnourished from weaning, presented decreased total and haploid cells in the vas deferens, hypertrophy of the muscle layer in the epididymal portion of the vas deferens and intense atrophy of the muscular coat in its prostatic portion. At a molecular level, the vas deferens tissue of CM rats exhibited a huge rise in lipid peroxidation and protein carbonylation, evidence of an accentuated increase in local reactive oxygen species levels. The kinetics of plasma membrane Ca2+-ATPase activity and its kinase-mediated phosphorylation by PKA and PKC in the vas deferens revealed malnutrition-induced modifications in velocity, Ca2+ affinity and regulation of Ca2+ handling proteins. The severely crippled content of the 12-kDa FK506 binding protein, which controls passive Ca2+ release from the sarco(endo) plasmic reticulum, revealed another target of malnutrition related to intracellular Ca2+ handling, with a potential effect on forward propulsion of sperm cells. As a possible compensatory response, malnutrition led to enhanced sarco(endo) plasmic reticulum Ca2+-ATPase activity, possibly caused by stimulatory PKA-mediated phosphorylation.Conclusions/SignificanceThe functional correlates of these cellular and molecular hallmarks of chronic malnutrition on the vas deferens were an accentuated reduction in fertility and fecundity.

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Section: Introductionmentioning

confidence: 99%

Undernutrition Affects Cell Survival, Oxidative Stress, Ca2+ Handling and Signaling Pathways in Vas Deferens, Crippling Reproductive Capacity

et al. 2013

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“…Taken together, alterations in vas are clinically applicable, and may have implications for fertility, ejaculation abnormalities, and possibly vasectomy associated pain (Granitsiotis and Kirk, 2004; Tandon and Sabanegh, 2008). Posed mechanisms for this supersensitivity are thought to be denervation related, and range from increased receptor density (Hata et al, 1981), partial resting membrane potential depolarization (Fleming et al, 1973; Fleming, 1975; Fleming and Westfall, 1975; Hershman et al, 1992, 1993, 1995) and changes in intracellular secondary messenger transduction and calcium sensitivity (Minneman et al, 1988; Abraham et al, 2003; Quintas et al, 2005; Amobi et al, 2006). …”

Section: Signal Transductionmentioning

confidence: 99%

Physiological and pharmacological aspects of the vas deferens—an update

Koslov¹,

Andersson²

2013

Front. Pharmacol.

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The vas deferens, a muscular conduit conveying spermatozoa from the epididymis to the urethra, has been used as a model tissue for smooth muscle pharmacological and physiological advancements. Many drugs, notably α-adrenergic antagonists, have effects on contractility and thus normal ejaculation, incurring significant side effects for patients that may interfere with compliance. A more thorough understanding of the innervation and neurotransmitter pharmacology of the vas has indicated that this is a highly complex structure and a model for co-transmission at the synapse. Recent models have shown clinical scenarios that alter the vas contraction. This review covers structure, receptors, neurotransmitters, smooth muscle physiology, and clinical implications of the vas deferens.

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“…Values from equilibrium binding experiments (dissociation constant, K d , and maximal number of binding sites, B max ) were calculated assuming the existence of one class of specific binding sites in the range of concentrations used, by means of computerized non-linear regression analysis of the untransformed data (Prism 4.0, GraphPad Software Inc., USA), as reported earlier (28). Statistical analysis for comparing transgenic to wild-type values was performed using the unpaired Student t-test (P < 0.05).…”

Section: Discussionmentioning

confidence: 99%

“…Subsequently, membranes were incubated for 1 h at room temperature with one of the following primary antibodies: monoclonal mouse IgG anti-Na + /K + -ATPase α 1 (McK1) and α 2 (McB2) isoforms (Dr. K. Sweadner, Harvard Medical School, Boston, MA, USA), anti-PMCA (clone 5F10, Sigma Chemical, USA), anti-SERCA2 (Calbiochem, EMD Chemicals, USA) or polyclonal rabbit anti-total or phospho-MAPKs antibodies (ERK1/2, p38 or SAPK/JNK; Cell Signaling Technology, USA) and for 1 h with horseradish peroxidaseconjugated anti-mouse (ATPases) or anti-rabbit (MAPKs) antibody (Promega, USA). Proteins were detected using an ECL detection kit (GE Healthcare Life Sciences, USA) and blot images were captured with a Bio-Rad GS-700 Imaging Densitometer and analyzed using the Quantity One Imaging software (Bio-Rad Laboratories, USA) (25,28). Loading control was performed by densitometric examination of Ponceau red staining as well as total MAPK blotting.…”

Section: Immunoblotting Assaysmentioning

confidence: 99%

“…Ca 2+ -ATPase activity was calculated by subtracting the basal ATPase activity (measured in the absence of Ca 2+ ) from the total ATPase activity (measured in the presence of 10 µM free Ca 2+ , with or without 3 µM thapsigargin; Sigma-Aldrich). The thapsigargin-resistant Ca 2+ -ATPase activity (assumed to be due to PMCA pumps) was subtracted from the total Ca 2+ -ATPase activity to obtain the thapsigargin-sensitive Ca 2+ -ATPase activity (assumed to be due to SERCA pumps) (28).…”

Section: Ca 2+ -Atpase Activitymentioning

confidence: 99%

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Lack of evidence for regulation of cardiac P-type ATPases and MAP kinases in transgenic mice with cardiac-specific overexpression of constitutively active α1B-adrenoceptors

Barreto

Rezende

Scaramello

et al. 2010

Braz J Med Biol Res

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The regulatory function of α 1B -adrenoceptors in mammalian heart homeostasis is controversial. The objective of the present study was to characterize the expression/activity of key proteins implicated in cardiac calcium handling (Na + /K + -ATPase and Ca 2+ -ATPases) and growth (ERK1/2, JNK1/2 and p38) in mice with cardiac-selective overexpression of constitutively active mutant α 1B -adrenoceptor (CAMα 1B -AR), which present a mild cardiac hypertrophy phenotype. Immunoblot assays showed that myocardial plasma membrane Ca 2+ -ATPase (PMCA) expression was increased by 30% in CAMα 1B -AR mice (N = 6, P < 0.05), although there was no change in sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA2) expression. Moreover, total Ca 2+ -ATPase activity was not modified, but a significant increase in the activity of the thapsigargin-resistant (PMCA) to thapsigargin-sensitive (SERCA) ratio was detected. Neither Na + /K + -ATPase activity nor the expression of α 1 and α 2 subunit isoforms was changed in CAMα 1B -AR mouse hearts. Moreover, immunoblot assays did not provide evidence for an enhanced activation of the three mitogen-activated protein kinases studied in this stage of hypertrophy. Therefore, these findings indicate that chronic cardiac α 1B -AR activation in vivo led to mild hypertrophy devoid of significant signs of adaptive modifications concerning primary intracellular calcium control and growth-related proteins, suggesting a minor pathophysiological role of this adrenergic receptor in mouse heart at this stage of development.

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Adaptive expression pattern of different proteins involved in cellular calcium homeostasis in denervated rat vas deferens

Cited by 12 publications

References 34 publications

Undernutrition Affects Cell Survival, Oxidative Stress, Ca2+ Handling and Signaling Pathways in Vas Deferens, Crippling Reproductive Capacity

Undernutrition Affects Cell Survival, Oxidative Stress, Ca2+ Handling and Signaling Pathways in Vas Deferens, Crippling Reproductive Capacity

Physiological and pharmacological aspects of the vas deferens—an update

Lack of evidence for regulation of cardiac P-type ATPases and MAP kinases in transgenic mice with cardiac-specific overexpression of constitutively active α1B-adrenoceptors

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