scholarly journals Acrosome reaction-inducing substance triggers two different pathways of sperm intracellular signaling in newt fertilization

2019 ◽  
Vol 63 (11-12) ◽  
pp. 589-595
Author(s):  
Shinnosuke Kon ◽  
Akio Takaku ◽  
Fubito Toyama ◽  
Eriko Takayama-Watanabe ◽  
Akihiko Watanabe
Keyword(s):  
Intracellular Signaling ◽  
Acrosome Reaction ◽  
Sperm Quality ◽  
Adenylyl Cyclases ◽  
Chain Reaction ◽  
Gated Channel ◽  
Polymerase Chain ◽  
Intracellular Pathways ◽  
Intracellular Mediators ◽  
Kinase A

The acrosome reaction is induced in the sperm of Cynops pyrrhogaster immediately in response to a ligand protein called acrosome reaction-inducing substance (ARIS) in the egg jelly at fertilization, whereas a spontaneous acrosome reaction occurs time-dependently in correlation with the decline of sperm quality for fertilization. The ARIS-induced acrosome reaction was recently found to be mediated by TRPV4 in association with the NMDA type glutamate receptor, although the intracellular mediators for the acrosome reaction are largely unclear. In the present study, spontaneous acrosome reaction was significantly inhibited by Ni2+, RN1734, and diltiazem, which blocks Cav3.2, TRPV4 or TRPM8, and the cyclic nucleotide-gated channel, respectively. In contrast, expression of Ca2+-activated transmembrane and soluble adenylyl cyclases was detected in the sperm of C. pyrrhogaster by reverse transcription-polymerase chain reaction. Activator of transmembrane or soluble adenylyl cyclases (forskolin or HCO3-) independently promoted spontaneous acrosome reaction, while an inhibitor of each enzyme (MD12330A or KH7) inhibited it only in the sperm with high potential for spontaneous acrosome reaction. An inhibitor of protein kinase A (H89) inhibited spontaneous acrosome reaction in a manner independent of sperm potential for spontaneous acrosome reaction. Surprisingly, KH7 significantly inhibited ARIS-induced acrosome reaction, but its effect was seen in a small percentage of sperm. H89 had no effect on ARIS-induced acrosome reaction. These results suggest that C. pyrrhogaster sperm possess multiple intracellular pathways for acrosome reaction, involving Ca2+ permeable channels, adenylyl cyclases and PKA, and that two pathways having distinct dependencies on adenylyl cyclases may contribute to ARIS-induced acrosome reaction at fertilization.

Molecular Pathology of Soft Tissue and Bone Tumors

1999 ◽  
Vol 123 (12) ◽  
pp. 1246-1259
Author(s):  
Andrzej Slominski ◽  
Jacobo Wortsman ◽  
Andrew Carlson ◽  
Martin Mihm ◽  
Brian Nickoloff ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Growth Factors ◽  
Soft Tissue ◽  
Reverse Transcription ◽  
Intracellular Signaling ◽  
Bone Tumors ◽  
Cellular Proliferation ◽  
Hybridization Technique ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Objective.—To present recent concepts on the molecular pathogenesis of tumors of soft tissue and bone, and on the use of molecular genetic methods, including their significance as diagnostic markers and prognostic indicators. Data Sources and Study Selection.—Reports on tumors of bone and/or soft tissue published in the English language literature and observations made using specimens available at the Departments of Pathology at Albany Medical College and Loyola University Medical Center. Data Extraction and Synthesis.—Studies on bone and soft tissue tumors containing chromosomal or genetic evaluation were selected for further analysis. Specific chromosomal abnormalities, such as numerical aberrations or translocations with production of fusion genes, were classified according to the tumor of origin. Data were also collected on mutations in tumor suppressor genes, genes coding for growth factors or their receptors, and genes coding for tyrosine kinases. Also noted were mutations of uncertain significance, for which the pathogenic connection between tumor production and mutated gene function is still unclear. Conclusions.—In general, the mutations reported interfere with the action of peptide growth factors coordinating mesenchyme proliferation and differentiation, although membrane-bound receptors expressing the intracellular signaling modifier, tyrosine kinase activity, have also been involved. Functional types of genes most commonly affected include tumor suppressors, oncogenes, and nuclear transcription factors. Thus, the mutations involved in the pathogenesis of soft tissue and bone tumors have affected multiple genes. Moreover, aberrant fusion gene products may be formed in tumoral tissue and may then act as transcription regulators stimulating cellular proliferation. Cytogenetic studies help at the clinical level by demonstrating aneuploidy and increased ploidy, which may correlate with malignant behavior. Diagnostic tumor-specific chromosomal translocations may be detected with Southern hybridization analysis, polymerase chain reaction, reverse-transcription polymerase chain reaction, or with the fluorescence in situ hybridization technique. Notably, early metastatic disease may be detectable in blood specimens using polymerase chain reaction or reverse-transcription polymerase chain reaction techniques.

Highly Conserved Sequence of Exon 15 BRAF Gene and KRAS Codon 12 Mutation among Greek Patients with Colorectal Cancer

2007 ◽  
Vol 22 (1) ◽  
pp. 12-18 ◽  
Author(s):  
E.K. Symvoulakis ◽  
A. Zaravinos ◽  
D. Panutsopulos ◽  
O. Zoras ◽  
E. Papalambros ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Intracellular Signaling ◽  
Mutational Analysis ◽  
Direct Sequencing ◽  
Chain Reaction ◽  
Braf Mutations ◽  
Conserved Sequence ◽  
Polymerase Chain ◽  
Kras Codon

Background The RAS/RAF/MEK/MAP kinase pathway is essential to intracellular signaling transduction regulating cell proliferation, differentiation and death. We investigated the occurrence of exon 15 BRAF and KRAS codon 12 mutations among Greek patients with colorectal cancer. Methods Sixty-one samples from patients with sporadic colorectal adenocarcinomas were studied for exon 15 BRAF mutations. DNA from surgically resected specimens was analyzed by a combination of polymerase chain reaction and direct sequencing. KRAS codon 12 mutational analysis was technically possible in 58 samples (58/61) by a combination of polymerase chain reaction and restriction fragment length polymorphism. Results No exon 15 BRAF mutations were detected in any of the colon cancer specimens. The frequency of KRAS codon 12 mutations was 29.3% (17/58). Patients aged <70 years more frequently presented carcinomas harboring KRAS codon 12 mutations than patients aged >70 years (p=0.028). Patients between 61 and 70 years of age were more likely to be carriers of this mutation (p=0.040). Conclusions Despite the limited study sample, our data suggest that BRAF mutations might be present less frequently than KRAS mutations in Greek patients with colorectal carcinomas. Further research involving larger patient series will be necessary to confirm these findings and to assess possible ethnic, environmental and lifestyle influences on BRAF and KRAS mutagenesis.

scholarly journals Molecular Characterization of Microbiota Associated With Sperm of Malaysian Mahseer Tor tambroides

2019 ◽  
Vol 15 ◽  
pp. 117693431985082
Author(s):  
Ivan Chong Chu Koh ◽  
Bin Hassan Badrul Nizam ◽  
Yazid Muhammad Abduh ◽  
Ambok Bolong Abol Munafi ◽  
Shumpei Iehata
Keyword(s):  
Gel Electrophoresis ◽  
Pathogenic Bacteria ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Sperm Quality ◽  
Good Prospect ◽  
Chain Reaction ◽  
Gradient Gel Electrophoresis ◽  
Polymerase Chain ◽  
Staphylococcus Spp

Malaysian Mahseer ( Tor tambroides) is considered as a good prospect for aquaculture in Malaysia. However, knowledge about Malaysian Mahseer-associated sperm microbiota is still limited, although some studies reported that sperm-related bacteria are a factor in the decline of sperm quality, as sperm may become the carrier of pathogenic bacteria to the egg. The goal of this study was to evaluate the sperm microbiota associated with Malaysian Mahseer from 3 different locations (Universiti Malaysia Terengganu [UMT], Ajil, and Pahang) using polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) fingerprinting and to compare location differences by cluster analysis. Our results showed that the UMT sample had different sperm microbiota composition and a different trend in its relationship with sperm quality. Correlation analysis showed a relationship between bacterial diversity and sperm quality. Phylogenetic analysis indicated that sperm microbiota was composed of diverse phyla, including Proteobacteria, Firmicutes, and Actinobacteria. Interestingly, bacteria such as Salinisphaera sp., Pelomonas sp., and Staphylococcus spp. were detected in all the locations, suggesting that these bacteria are indigenous bacterial members of the Malaysian Mahseer sperm microbiota, although their function is still unclear.

scholarly journals Polymorphism of follicle stimulating hormone beta sub-unit (FSH-P) gene as a molecular marker for reproductive status in Peranakan Ongole x Bali crossbred (POBA) cattle

2021 ◽  
Vol 902 (1) ◽  
pp. 012052
Author(s):  
A Primasari ◽  
J Efendy ◽  
P W Prihandini
Keyword(s):  
Sperm Quality ◽  
Follicle Stimulating Hormone ◽  
Reproductive Status ◽  
Crossbred Cattle ◽  
Phenotypic Characteristics ◽  
Chain Reaction ◽  
P Gene ◽  
Pcr Products ◽  
Polymerase Chain ◽  
The Republic

Abstract POBA cattle is a crossbred cattle from Peranakan Ongole (PO) and Bali cattle, which is developed by the Beef Cattle Research Institute (BCRI), Ministry of Agriculture of the Republic of Indonesia. This study aimed to identify the genetic and phenotypic characteristics of POBA cattle raised in the BCRI. Genetic diversity of FSH-β gene was also determined in order to identify a possible marker for reproductive status in POBA cattle. Rambon cattle collected from Banyuwangi regency were used as comparison. A 313 bp fragment of the FSH-β gene was amplified using the polymerase chain reaction (PCR). The PCR products were sequenced and aligned to detect polymorphism. As results, a polymorphism (SNP g.2583C/T) in the FSH-β gene, which produced three genotypes (TT, CC, and CT) was detected. The frequency of each allele was 0.13 (allele C) and 0.87 (allele T). However, the FSH-β gene polymorphism did not significantly affect sperm quality, body weight, body size, and cervical condition of POBA cattle.

Detection of glycine receptor/Cl- channel beta subunit transcripts in mouse testis

Zygote ◽  
2002 ◽  
Vol 10 (2) ◽  
pp. 105-108 ◽  
Author(s):  
Yoko Sato ◽  
Richard P. Tucker ◽  
Stanley Meizel
Keyword(s):  
Acrosome Reaction ◽  
Glycine Receptor ◽  
Beta Subunit ◽  
Seminiferous Tubules ◽  
Chain Reaction ◽  
Mammalian Sperm ◽  
Hybridisation Analysis ◽  
Polymerase Chain ◽  
Sperm Acrosome Reaction

The sperm glycine receptor/Cl- channel (GlyR) is important to the initiation of the mammalian sperm acrosome reaction by the egg zona pellucida, but its presence in spermatogenic cells has not been demonstrated. Reverse transcriptase-polymerase chain reaction studies confirmed that GlyR beta subunit transcripts similar to those of the neuronal GlyR beta subunit are present in the testis of Swiss Webster mice. In situ hybridisation analysis demonstrated that GlyR beta subunit mRNAs were expressed within the germ cells of seminiferous tubules in those mice.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain
Sign in / Sign up

Export Citation Format

Share Document