Abstract
Turtles grow slowly and have a long lifespan. Ultrastructural studies of the pituitary gland in Reeves’ turtle (Chinemys reevesii) have revealed that the species possesses a higher nucleoplasmic ratio and fewer secretory granules in growth hormone (GH) cells than other animal species in summer and winter. C. reevesii GH gene was cloned and species-specific similarities and differences were investigated. The full GH gene sequence in C. reevesii contains 8517 base pairs (bp), comprising five exons and four introns. Intron 1 was found to be much longer in C. reevesii than in other species. The coding sequence (CDS) of the turtle’s GH gene, with and without the inclusion of intron 1, was transfected into four cell lines, including DF-1 chicken embryo fibroblasts, Chinese hamster ovary (CHO) cells, human embryonic kidney 293FT cells, and GH4C1 rat pituitary cells; the turtle growth hormone (tGH) gene mRNA and protein expression levels decreased significantly in the intron-containing CDS in these cell lines, compared with that of the corresponding intronless CDS. Thus, the long intron 1 of GH gene in Reeves’ turtle might correlate with downregulated gene expression.
Highlights
Reeves’ turtle (Chinemys reevesii, known as Mauremys reevesii) belongs to the order Testudinata in the family Bataguridae and is found extensively in China, Japan, Korea, and other parts of Asia
We found that long intron 1 in the growth hormone (GH) gene can have both positive and negative effects on GH gene expression, and there is evidence to support the hypothesis that this intron may directly regulate GH expression
There have been no data presented on the function of intron 1 in turtles of this or any other species in vivo or in vitro before
Summary
Reeves’ turtle (Chinemys reevesii, known as Mauremys reevesii) belongs to the order Testudinata in the family Bataguridae and is found extensively in China, Japan, Korea, and other parts of Asia. In the current study, we cloned the GH gene of this species and investigated specific similarities and differences in its sequence using four cell lines, including DF-1 chicken embryo fibroblasts, Chinese hamster ovary (CHO) cells, human embryonic kidney 293FT cells, and GH4C1 rat pituitary cells. In these cell lines, we compared the expression of a GH intron 1-containing coding sequence (CDS) with that of a corresponding intronless CDS. We found that long intron 1 in the GH gene can have both positive and negative effects on GH gene expression, and there is evidence to support the hypothesis that this intron may directly regulate GH expression
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