Molecular characterization of the leopard gecko POMC gene and expressional change in the testis by acclimation to low temperature and with a short photoperiod.
The gene for proopiomelanocortin (POMC), a common precursor of malanotropins, corticotropin, and beta-endorphin, was isolated and analyzed in the squamata species, the leopard gecko, Eublepharis macularius. Leopard gecko POMC (lgPOMC) cDNA is composed of 1299bp, excluding the poly(A) tail, and encodes 270 amino acids. The deduced amino acid sequence showed the same structural organization as that of other species and displayed identity with those of other vertebrates: 68% with mud turtles, 57/57% with African clawed frog A/B, 53% with chickens, and 45% with mice. In a phylogenic tree, the lgPOMC clustered with the sequences of the mud turtle POMC and python POMC. The lgPOMC gene comprises three exons and two introns and this structure is consistent with humans, rats, mice, African clawed frog and zebrafish. RT-PCR analysis revealed that the lgPOMC mRNA was expressed only in the whole brain, pituitary, and gonads. To analyze in more detail, a competitive assay system to quantify the expression levels of POMC mRNA was established. We measured the POMC mRNA expression levels in the leopard gecko testes following transfer from a condition of 29 degrees C, 16L/8D to 18 degrees C, 10L/14D over 6 weeks. This 6-week acclimation increased the POMC mRNA expression levels significantly. This suggests that the leopard gecko POMC-derived peptides play a role in the mediation of the effect of environmental factors on reproduction.
Endo
Daisuke
D
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033, Japan. MS7D-ENDU@asahi-net.or.jp
Park
Min Kyun
MK
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003080:Cold Temperature; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D008116:Lizards / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D017440:Photoperiod; D010802:Phylogeny; D011333:Pro-Opiomelanocortin / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D013737:Testis / Q000378:metabolism*
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D011333:Pro-Opiomelanocortin