Molecular cloning of several rat ABC transporters including a new ABC transporter, Abcb8, and their expression in rat testis.
Several members of the ABC transporter superfamily play an important role in testicular physiology and defence against anticancer drugs. Using a reverse transcription-polymerase chain reaction strategy with degenerate primers and rat testis RNA as template, we have looked for the presence of other members of this superfamily. Of the six partial cDNA found, five corresponded to ABC transporters already known -Mdr1b, Mrp1, Tapl/Abcb9, Umat/Abcb6 and Sur2/Abcc9- and one presented a strong homology with mouse and human ABCB8. Using a 5' and 3' RACE approach, we cloned the full-length cDNA and found that the predicted protein presented 92% and 80% homology with the mouse and human proteins respectively. Strong expression of rat abcb8 was found in heart, brain and testis when compared with liver, lung and spleen. In the testis, rat abcb8 was expressed both in the somatic Sertoli cells and peritubular cells and in the germline (spermatogonia and pachytene spermatocytes). Furthermore, Umat/Abcb6 was very highly expressed in the testis (high amounts in meiotic pachytene spermatocytes and low amount in post-meiotic early spermatids). In conclusion, we confirm the presence of several ABC transporters in the testis and also provide evidence of the presence of Abcb8 in the organ.
Melaine
Nathalie
N
GERHM-INSERM U625, Campus de Beaulieu, Université de Rennes I, Rennes Cedex, Bretagne, France.
Satie
Anne-Pascale
AP
Lassurguère
Julie
J
Desmots
Sophie
S
Jégou
Bernard
B
Samson
Michel
M
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000502:physiology; D003001:Cloning, Molecular; D017931:DNA Primers; D015870:Gene Expression; D006321:Heart / Q000502:physiology; D007985:Leydig Cells / Q000502:physiology; D008099:Liver / Q000502:physiology; D008168:Lung / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D024661:Potassium Channels, Inwardly Rectifying / Q000235:genetics; D051381:Rats; D011955:Receptors, Drug; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012708:Sertoli Cells / Q000502:physiology; D013087:Spermatids / Q000502:physiology; D013093:Spermatogonia / Q000502:physiology; D013154:Spleen / Q000502:physiology; D064233:Sulfonylurea Receptors; D013737:Testis / Q000166:cytology / Q000502:physiology*
C577510:ABCC9 protein, human; D018528:ATP-Binding Cassette Transporters; C491540:Abcb6 protein, rat; C512027:Abcb8 protein, rat; C577512:Abcc9 protein, rat; D017931:DNA Primers; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors