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

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.

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Identification and spatial distribution of glycine receptor subunits in human sperm

Reproduction ◽

10.1530/rep-08-0223 ◽

2008 ◽

Vol 136 (4) ◽

pp. 387-390 ◽

Cited By ~ 5

Author(s):

Priyadarsini Kumar ◽

Stanley Meizel

Keyword(s):

Spatial Distribution ◽

Western Blot ◽

Blot Analysis ◽

Acrosome Reaction ◽

Glycine Receptor ◽

Human Sperm ◽

Equatorial Region ◽

Mammalian Sperm ◽

Principal Piece ◽

Sperm Acrosome Reaction

The human sperm surface glycine receptor (GLR) plays a role in an important fertilization event, the sperm acrosome reaction. Here, by western blot analysis, we report the presence of GLRA1, GLRA2, GLRA3, and GLRB subunits in human sperm. Immunolocalization studies showed that the GLRA1 and GLRA2 subunits are present in the equatorial region, the GLRA3 subunit in the flagellar principal piece, and the GLRB subunit in the acrosomal region of sperm. This first demonstration of isoforms of the sperm GLRA subunit and of a differential spatial distribution of the α and β subunits on the surface of mammalian sperm suggests the possibility that human sperm GLRs have more than one function.

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

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100166944 ◽

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.

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