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  • Author: B Funkenstein x
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M Khayat, E Lubzens, A Tietz, and B Funkenstein

ABSTRACT

A cDNA clone encoding a female-specific ovarian protein (presumably vitellin, Vt) has been isolated from a cDNA library prepared from poly (A)+ RNA extracted from vitellogenic ovaries of the shrimp Penaeus semisulcatus. The cDNA library was constructed and screened using a major cDNA band which was observed following analysis of total cDNA products by gel electrophoresis. This band, as well as the cDNA insert purified from the library, was estimated to have 1.1 kb. Both hybridized to mRNA prepared from ovaries or hepatopancreas (HEP) of vitellogenic females and showed a faint signal with ovaries from non-vitellogenic females, but did not hybridize to HEP from non-vitellogenic females or to HEP from males or testes. The size of the transcripts from the ovary and HEP was estimated to be 1.1 kb, similar to that of the cDNA insert, suggesting that a full length cDNA had been synthesized. Furthermore, the identical sizes of the transcripts from ovary and HEP and the ability of the ovarian cDNA to detect a transcript in HEP mRNA suggest that Vt from the ovary and vitellogenin (Vg) from HEP are the gene products of one gene. Alternatively, the homology between Vt and Vg is very high.

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S J Duguay, J Lai-Zhang, D F Steiner, B Funkenstein, and S J Chan

ABSTRACT

Recent studies have shown that homologues of the mammalian IGF-I and -II genes are also found in teleosts. We report here the cDNAs coding for IGF-I and IGF-II cloned from the gilthead seabream, Sparus aurata. Sequence comparisons revealed that both IGFs have been well conserved among teleosts, although Sparus IGF-I is shorter by three amino acid residues due to truncated B-and C-domains. Using the cloned cDNAs as probes, the relative expression of IGF-I and IGF-II mRNAs were assayed in different Sparus tissues. Sparus liver clearly contained the highest level of IGF-I mRNA while relatively high levels of IGF-II mRNA were found in liver, heart and gill using the ribonuclease protection assay. After GH administration the amount of IGF-I mRNA was increased by 220% in liver but no changes in IGF-II mRNA levels were detected in any tissue. We also assayed the expression of IGF-I and IGF-II in Sparus during early development. The IGF-II mRNA level was highest in larva 1 day after hatching and decreased thereafter. In contrast, IGF-I mRNA was detected in 1-day-old larva but there was an increase in expression in 12- and 16-day-old larva. These results demonstrated that the expression of IGF-I and IGF-II is highly regulated in teleosts and suggest that they play distinct roles during growth and development.