Abstract
To discover late stage germ cell-specific transcripts we prepared a cDNA library from adult testes of 35-day old mice and subtracted it with mRNA from the testes of juvenile mice. Real-time RT-PCR analysis indicated that 42 cDNA clones in the subtracted library were expressed more intensely in the adult testes than in the juvenile testes. One clone identified by subtraction is expressed preferentially in the late spermatid and is located on chromosome 17E3 in mouse and 2p22 in human. The full nucleotide and amino acid sequences of mouse and human MOPT gene are deposited in EMBL GenBank (AY367765 And AY367766). Human MOPT is spliced by 5 exons and 4 introns and encompasses 7,000 bp of genomic DNA (from bp 355 822 to 425 511) of NT-022184.13, whereas mouse MOPT is spliced by 5 exons and 4 introns and encompasses 7,382 bp of genomic DNA (from bp 6227407 to 6235588) of NT-039658.2. Because of the limited availability of human testis samples, development-dependent expression of MOPT mRNA was conducted using its mouse homologue and semiquantitative PCR. The number of cycles completed before entering the exponential growth, recorded by amplifier PE5700 for mouse MOPT, were 1.11 ± 0.23, 1.05 ± 0.04, 1.5 ± 0.2, 5.55 ± 0.65, 19.35 ± 0.65, 68.65 ± 2.15, and 185.15 ± 6.15 in W/W, postnatal day 5-, 8-, 12-, 15-, 18-, 22-, and 28-day mouse tissue samples, respectively. The difference among the three times was significant (P < 0.01, ANOVA). These results suggest that expression of MOPT gene increased from postnatal Day 5 to Day 28, indicating possible involvement in testicular development. The ORF encodes a protein containing 79 amino acid residues. A MORN motif, EGQFKDNMFHGLGTYTFPNG, was identified in the predicted protein sequence of MOPT; function of this motif is unknown. In situ hybridization of 12-week-old wild-type mouse testes using an antisense riboprobe and immuno-gold data indicated MOPT was expressed as a late spermatid and acrosome reaction. This work was supported by BK21 program.
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