Hormonally-regulated Differential Expression of the Two Duplicated Insecticyanin Genes, Ins-a and Ins-b, During Development of the Tobacco Hornworm, Manduca Sexta

Overview

Journal Rouxs Arch Dev Biol

Specialty Biology

Date 2017 Mar 18

PMID 28306068

Authors

Wan-Cheng Li

Kiyoshi Hiruma

Lynn M Riddiford

Affiliations

Soon will be listed here.

Abstract

The effects of juvenile hormone (JH) and 20-hydroxyecdysone (20E) on the developmental expression of the two insecticyanin genes, ins-a and ins-b, were investigated with two gene-specific probes. Removal of the corpora allata (-CA, source of JH) clearly delayed and down-regulated the epidermal expression of these genes but enhanced their expression in the fat body during the early development of the fifth instar. Application of JH I to the -CA larvae at the time of head capsule slippage completely restored the normal epidermal expression pattern of the two genes in the early fifth instar, then INS-a mRNA declined prematurely whereas INS-b mRNA remained similar to that in the intact larvae. By contrast, in the fat body of -CA larvae, the exogenous JH had little effect on the levels of INS-a mRNA, but enhanced expression of INS-b mRNA relative to intact larvae. Culture of epidermis from day 1 fifth instar larvae with 40 ng/ml 20E for up to 24 h accelerated the loss of INS-a mRNA without affecting the levels of INS-b mRNA. Both mRNAs declined in isolated larval abdomens over a 24 h period, and this decline was slowed by 1 μg methoprene (a JH analog). Together these results indicate that JH controls the levels of the two mRNAs in both the epidermis and fat body, with additional factors involved in regulating these genes in the fat body during the molt and in the epidermis during the growth phase.

References

Denhardt D . A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966; 23(5):641-6. DOI: 10.1016/0006-291x(66)90447-5. View

Li W, Riddiford L . The two duplicated insecticyanin genes, ins-a and ins-b are differentially expressed in the tobacco hornworm, Manduca sexta. Nucleic Acids Res. 1994; 22(15):2945-50. PMC: 310259. DOI: 10.1093/nar/22.15.2945. View

Li W, Riddiford L . Two distinct genes encode two major isoelectric forms of insecticyanin in the tobacco hornworm, Manduca sexta. Eur J Biochem. 1992; 205(2):491-9. DOI: 10.1111/j.1432-1033.1992.tb16805.x. View

Goodman W, Adams B, Trost J . Purification and characterization of a biliverdin-associated protein from the hemolymph of Manduca sexta. Biochemistry. 1985; 24(5):1168-75. DOI: 10.1021/bi00326a017. View

Law J, Wells M . Insects as biochemical models. J Biol Chem. 1989; 264(28):16335-8. View

Riddiford L, Palli S, Hiruma K, Li W, Green J, Hice R . Developmental expression, synthesis, and secretion of insecticyanin by the epidermis of the tobacco hornworm, Manduca sexta. Arch Insect Biochem Physiol. 1990; 14(3):171-90. DOI: 10.1002/arch.940140305. View

Riley C, Barbeau B, Keim P, Kezdy F, Heinrikson R, Law J . The covalent protein structure of insecticyanin, a blue biliprotein from the hemolymph of the tobacco hornworm, Manduca sexta L. J Biol Chem. 1984; 259(21):13159-65. View

Thomas P . Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980; 77(9):5201-5. PMC: 350025. DOI: 10.1073/pnas.77.9.5201. View

Feinberg A, Vogelstein B . "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984; 137(1):266-7. DOI: 10.1016/0003-2697(84)90381-6. View

10.

Fain M, Riddiford L . Juvenile hormone titers in the hemolymph during late larval development of the tobacco hornworm, Manduca sexta (L.). Biol Bull. 1975; 149(3):506-21. DOI: 10.2307/1540383. View

11.

Hiruma K, Riddiford L . Regulation of melanization of tobacco hornworm larval cuticle in vitro. J Exp Zool. 1984; 230(3):393-403. DOI: 10.1002/jez.1402300308. View

12.

Wolfgang W, Riddiford L . Larval cuticular morphogenesis in the tobacco hornworm, Manduca sexta, and its hormonal regulation. Dev Biol. 1986; 113(2):305-16. DOI: 10.1016/0012-1606(86)90166-1. View

13.

Dominick O, Truman J . The physiology of wandering behaviour in Manduca sexta. II. The endocrine control of wandering behaviour. J Exp Biol. 1985; 117:45-68. DOI: 10.1242/jeb.117.1.45. View

14.

Alwine J, Kemp D, Stark G . Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977; 74(12):5350-4. PMC: 431715. DOI: 10.1073/pnas.74.12.5350. View

15.

Riddiford L . Ecdysone-induced change in cellular commitment of the epidermis of the tobacco hornworm, Manduca sexta, at the initiation of metamorphosis. Gen Comp Endocrinol. 1978; 34(4):438-46. DOI: 10.1016/0016-6480(78)90284-8. View

16.

Hiruma K . Possible roles of juvenile hormone in the prepupal stage of Mamestra brassicae. Gen Comp Endocrinol. 1980; 41(3):392-9. DOI: 10.1016/0016-6480(80)90084-2. View

17.

Kafatos F, Jones C, Efstratiadis A . Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979; 7(6):1541-52. PMC: 342326. DOI: 10.1093/nar/7.6.1541. View