Testicular Expression of Adora3i2 in Adora3 Knockout Mice Reveals a Role of Mouse A3Ri2 and Human A3Ri3 Adenosine Receptors in Sperm*

Burnett, Lindsey A.; Blais, Edik M.; Unadkat, Jashvant D.; Hille, Bertil; Tilley, Stephen L.; Babcock, Donner F.

doi:10.1074/jbc.m110.156075

Cited by 24 publications

(19 citation statements)

References 46 publications

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“…A strikingly similar attenuated flagellar beat and impaired sperm hyperactivation was also seen in sperm lacking the sperm-specific catalytic subunit of PKA, Cαs [25]. Flagellar beat waveform features seen with Gsk3a null sperm were observed in membrane permeabilized sperm activated in the presence of AMP [32] and also in sperm lacking the nucleoside transport protein Slc29a (ENT1) [33]. Further studies with sperm from Gsk3a (-/-) null mice are required where quantitation of flagellar beat amplitude and frequency can be coupled to biochemical changes in null sperm.…”

Section: Discussionmentioning

confidence: 89%

Targeted Disruption of Glycogen Synthase Kinase 3a (Gsk3a) in Mice Affects Sperm Motility Resulting in Male Infertility1

Bhattacharjee

Goswami

Dudiki

et al. 2015

Biology of Reproduction

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The signaling enzyme glycogen synthase kinase 3 (GSK3) exists as two isoforms-GSK3A and GSK3B. Protein phosphorylation by GSK3 has important signaling roles in several cells. In our past work, we found that both isoforms of GSK3 are present in mouse sperm and that catalytic GSK3 activity correlates with motility of sperm from several species. Here, we examined the role of Gsk3a in male fertility using a targeted gene knockout (KO) approach. The mutant mice are viable, but have a male infertility phenotype, while female fertility is unaffected. Testis weights of Gsk3a(-/-) mice are normal and sperm are produced in normal numbers. Although spermatogenesis is apparently unimpaired, sperm motility parameters in vitro are impaired. In addition, the flagellar waveform appears abnormal, characterized by low amplitude of flagellar beat. Sperm ATP levels were lower in Gsk3a(-/-) mice compared to wild-type animals. Protein phosphatase PP1 gamma2 protein levels were unaltered, but its catalytic activity was elevated in KO sperm. Remarkably, tyrosine phosphorylation of hexokinase and capacitation-associated changes in tyrosine phosphorylation of proteins are absent or significantly lower in Gsk3a(-/-) sperm. The GSK3B isoform was present and unaltered in testis and sperm of Gsk3a(-/-) mice, showing the inability of GSK3B to substitute for GSK3A in this context. Our studies show that sperm GSK3A is essential for male fertility. In addition, the GSK3A isoform, with its highly conserved glycine-rich N terminus in mammals, may have an isoform-specific role in its requirement for normal sperm motility and fertility.

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Section: Discussionmentioning

confidence: 89%

Targeted Disruption of Glycogen Synthase Kinase 3a (Gsk3a) in Mice Affects Sperm Motility Resulting in Male Infertility1

Bhattacharjee

Goswami

Dudiki

et al. 2015

Biology of Reproduction

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“…During isolation, each right cauda epididymis was rinsed and excised in phosphate buffered saline then minced in a 35-mm Petri dish containing 1.0 ml swimout/capacitation medium consisting of Medium HS [31] (in mM: 135 NaCl, 5 KCl, 2 CaCl2, 1 MgSO4, 20 HEPES, 5 glucose, 10 lactic acid, 1 pyruvic acid, pH 7.4) supplemented with 15 mM NaHCO 3 and 5 mg bovine serum albumin (BSA) (Fraction V)/ml preheated to 37 °C. The sperm-media suspension was incubated at 37 °C for 10 minutes.…”

Section: Methodsmentioning

confidence: 99%

Sub-acute intravenous administration of silver nanoparticles in male mice alters Leydig cell function and testosterone levels

Garcia

Costa

França

et al. 2014

Reproductive Toxicology

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The aim of this study was to determine whether short-term, in vivo exposure to silver nanoparticles (AgNPs) could be toxic to male reproduction. Low dose (1 mg/kg/dose) AgNPs were intravenously injected into male CD1 mice over 12 days. Treatment resulted in no changes in body and testis weights, sperm concentration and motility, fertility indices, or follicle-stimulating hormone and luteinizing hormone serum concentrations; however, serum and intratesticular testosterone concentrations were significantly increased 15 days after initial treatment. Histologic evaluation revealed significant changes in epithelium morphology, germ cell apoptosis, and Leydig cell size. Additionally, gene expression analysis revealed Cyp11a1 and Hsd3b1 mRNA significantly upregulated in treated animals. These data suggest that AgNPs do not impair spermatogonial stem cells in vivo since treatment did not result in significant decreases in testis weight and sperm concentrations. However, AgNPs appear to affect Leydig cell function, yielding increasing testicular and serum testosterone levels.

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“…Both cell lines are useful to demonstrate GPCR-triggered cAMP accumulation. However, unlike COS-7 cells, type II adenylyl cyclase I in TSA-201 cells is not stimulated by free βγ subunits released from activated Gα i protein which can inhibit cAMP accumulation when transfected with Gα i -coupled receptors such as CXCR1 [37], [38]. COS-7 cells and HEK293 cell-derived TSA-201 cells were maintained in DMEM, and supplemented with 10% FBS.…”

Section: Methodsmentioning

confidence: 99%

Leu1283.43 (L128) and Val2476.40 (V247) of CXCR1 Are Critical Amino Acid Residues for G Protein Coupling and Receptor Activation

et al. 2012

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CXCR1, a classic GPCR that binds IL-8, plays a key role in neutrophil activation and migration by activating phospholipase C (PLC)β through Gα15 and Gαi which generates diacylglycerol and inositol phosphates (IPs). In this study, two conserved amino acid residues of CXCR1 on the transmembrane domain (TM) 3 and TM6, Leu1283.43 (L128) and Val2476.40 (V247), respectively, were selectively substituted with other amino acids to investigate the role of these conserved residues in CXCR1 activation. Although two selective mutants on Leu128, Leu128Ala (L128A) and Leu128Arg (L128R), demonstrated high binding affinity to IL-8, they were not capable of coupling to G proteins and consequently lost the functional response of the receptors. By contrast, among the four mutants at residue Val247 (TM6.40), replacing Val247 with Ala (V247A) and Asn (V247N) led to constitutive activation of mutant receptors when cotransfected with Gα15. The V247N mutant also constitutively activated the Gαi protein. These results indicate that L128 on TM3.43 is involved in G protein coupling and receptor activation but is unimportant for ligand binding. On the other hand, V247 on TM6.40 plays a critical role in maintaining the receptor in the inactive state, and the substitution of V247 impaired the receptor constraint and stabilized an active conformation. Functionally, there was an increase in chemotaxis in response to IL-8 in cells expressing V247A and V247N. Our findings indicate that Leu1283.43 and Val2476.40 are critical for G protein coupling and activation of signaling effectors, providing a valuable insight into the mechanism of CXCR1 activation.

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Testicular Expression of Adora3i2 in Adora3 Knockout Mice Reveals a Role of Mouse A3Ri2 and Human A3Ri3 Adenosine Receptors in Sperm*

Cited by 24 publications

References 46 publications

Targeted Disruption of Glycogen Synthase Kinase 3a (Gsk3a) in Mice Affects Sperm Motility Resulting in Male Infertility1

Targeted Disruption of Glycogen Synthase Kinase 3a (Gsk3a) in Mice Affects Sperm Motility Resulting in Male Infertility1

Sub-acute intravenous administration of silver nanoparticles in male mice alters Leydig cell function and testosterone levels

Leu1283.43 (L128) and Val2476.40 (V247) of CXCR1 Are Critical Amino Acid Residues for G Protein Coupling and Receptor Activation

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