Identification and characterization of a novel splice variant of β3 subunit of GABA A receptor in rat testis and spermatozoa

Shi Feng Li, Jinghua Hu, Yuan Chang Yan, Yu Guang Chen, S. S. Koide, Yi Ping Li

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

γ-Aminobutyric acid type A (GABA A) receptors are the major sites of inhibitory action of fast synaptic neurotransmission in the brain. Their receptors are also widely distributed in peripheral and endocrine tissues. A full-length cDNA encoding a novel splice variant of β3 subunit of GABA A receptor, designated as β3t, was identified in rat testis. This isoform contains a segment, having identical amino acid sequence as the β3 subunit of neuronal GABA A receptors except for a section composed of 25 different amino acid sequence in the N-terminus. Northern blot shows that this isoform is found in rat testis. The β3t isoform mRNA was detected in germ cells in the late step of spermatogenesis by in situ hybridization assay. Results of immunohistochemical and immunocytochemical assays indicate that the β3t isoform is expressed in rat testis and spermatozoa. To determine a possible function of the N-terminal 25 amino acid segment, a recombinant plasmid of β3t-EGFPC was constructed by fusing green fluorescent protein to the C-terminus of the β3t isoform. The chimera product failed to be translocated unto the cell surface when expressed in HEK 293 cells; whereas, the β3 subunit of rat brain is incorporated into the plasma membrane. In conclusion, the present results show that one variant of β3 subunit of GABA A receptor, designated as β3t, is found in germ cells of rat testis and sperm. The inability of the β3t variant to target into the plasma membrane maybe a consequence of the unique 25 amino acid segment in the N-terminus.

Original languageEnglish (US)
Pages (from-to)350-360
Number of pages11
JournalInternational Journal of Biochemistry and Cell Biology
Volume37
Issue number2
DOIs
StatePublished - Feb 2005
Externally publishedYes

Fingerprint

GABA-A Receptors
Spermatozoa
Testis
Rats
Protein Isoforms
Amino Acids
Cell membranes
Germ Cells
Amino Acid Sequence
Assays
Brain
Cells
Cell Membrane
RNA Isoforms
Aminobutyrates
HEK293 Cells
Spermatogenesis
Protein C
Green Fluorescent Proteins
Synaptic Transmission

Keywords

  • β3 Subunit
  • GABA receptor
  • Rat testis and spermatozoa
  • Splice variant

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Identification and characterization of a novel splice variant of β3 subunit of GABA A receptor in rat testis and spermatozoa. / Li, Shi Feng; Hu, Jinghua; Yan, Yuan Chang; Chen, Yu Guang; Koide, S. S.; Li, Yi Ping.

In: International Journal of Biochemistry and Cell Biology, Vol. 37, No. 2, 02.2005, p. 350-360.

Research output: Contribution to journalArticle

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AB - γ-Aminobutyric acid type A (GABA A) receptors are the major sites of inhibitory action of fast synaptic neurotransmission in the brain. Their receptors are also widely distributed in peripheral and endocrine tissues. A full-length cDNA encoding a novel splice variant of β3 subunit of GABA A receptor, designated as β3t, was identified in rat testis. This isoform contains a segment, having identical amino acid sequence as the β3 subunit of neuronal GABA A receptors except for a section composed of 25 different amino acid sequence in the N-terminus. Northern blot shows that this isoform is found in rat testis. The β3t isoform mRNA was detected in germ cells in the late step of spermatogenesis by in situ hybridization assay. Results of immunohistochemical and immunocytochemical assays indicate that the β3t isoform is expressed in rat testis and spermatozoa. To determine a possible function of the N-terminal 25 amino acid segment, a recombinant plasmid of β3t-EGFPC was constructed by fusing green fluorescent protein to the C-terminus of the β3t isoform. The chimera product failed to be translocated unto the cell surface when expressed in HEK 293 cells; whereas, the β3 subunit of rat brain is incorporated into the plasma membrane. In conclusion, the present results show that one variant of β3 subunit of GABA A receptor, designated as β3t, is found in germ cells of rat testis and sperm. The inability of the β3t variant to target into the plasma membrane maybe a consequence of the unique 25 amino acid segment in the N-terminus.

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