MTR Gene Expression in Male Reproductive Tract Tissues
Sequence analysis of the various PCR fragments obtained using tammar MTR-specific oligonucleotide primers and various tissues produced a consensus nucleotide sequence of 588 bp, which encoded a peptide of 196 amino acids (Fig. 1). This derived amino acid sequence showed a high homology (74-77%) to the region extending from the II to the VI putative transmembrane domain in all eutherian OT receptor genes published but low homology to vasopressin receptors (38-52%). Of particular interest is the identification of highly conserved regions in the III transmembrane domain and the first and second extracellular loops (Fig. 1). Buy Advair Diskus Online
RT-PCR analysis revealed a single PCR product of 400 bp expressed in the prostate gland of the adult, juvenile, and pouch young and also in the epididymis of the adult and juvenile tammar (Fig. 2a). A strong positive signal was present in the myometrium of the late-pregnant female tam-mar. No MTR transcripts were observed in the testis. All samples showed strong positive signals for GAPDH mRNA (Fig. 2a), demonstrating equivalent quality of reverse transcription reactions.
FIG. 1. The partial 196-amino acid sequence of the putative tammar MTR cDNA. Areas of homology compared with the rat, bovine, human, and sheep OT receptor gene sequences are indicated with asterisks. The positions of the putative transmembrane domains are indicated above the arrows with Roman numerals. Of particular interest is the identification of highly conserved regions in the III transmembrane domain and in the first and second extracellular loops.
FIG. 2. a) MTR gene transcripts were detected in the prostate (P) and epididymis (E) of the adult (lanes 2 and 3), juvenile (lanes 5 and 6), and pouch young (PY: lane 7) and also in the myometrium (myo) of the late-pregnant female (lane 8). However, no MTR transcripts were detected in the testis (T) of either the adult or juvenile (lanes 1 and 4) or in the PCR reaction in which water replaced the cDNA template. All samples showed strong positive signals for GAPDH mRNA, demonstrating equivalent quality of reverse transcription reactions. MW: 100-bp DNA marker. b) Southern hybridization analysis of RT-PCR products specific for MTR mRNA. The membrane was hybridized with a radiolabeled tammar MTR oligonucleotide probe and exposed to film for 10 h. Control: Lane 8, where water replaced the cDNA in the PCR reaction.