Abstract
Temporal and spatial patterns in the accumulation of Tropoelastin (TE) mRNA during development of the chick embryo were established by in situ hybridization. Radiolabeled oligonucleotide probes of high specific activity were hybridized to serial sections of the cardiovascular system from embryonic day 3.5 (ED 3.5) to ED 19. Tropoelastin mRNA was observed as early as ED 3.5 in the dorsal part of the arterial trunk. During septation varying levels of TE mRNA were seen in the pulmonary trunk, the aorta and the aorticopulmonary septum. Thereafter TE mRNA levels increased up to ED 12, and the appearance of message was distributed distally in the walls of developing arteries. From ED 4.5 on, we found a decreasing proximo-distal gradient of the hybridization signal along the trunks and later along the main arteries (longitudinal gradient), and a radial gradient through the arterial vessel wall with the highest levels of TE mRNA in the outer layers of the media. Both gradients persisted in all major arterial vessels except in the proximal systemic and pulmonary trunks, where the original radial gradient was inverted or locally bimodal during the second half of development. The valvular region of aortic and pulmonary trunks showed particularly striking patterns of TE mRNA distribution, notably a prominent label on the endothelial cell layer on aortic and pulmonary valves. Outside the cardiovascular system, TE mRNA was mainly present in prochondral or perichondral cells in trachea and growing skeleton, and in the gap of growing joints. In kidney or nephric primordia, TE mRNA was only detectable in the wall of renal arteries. A hybridization signal was observed on mesenchyme of pulmonary septae at ED 16. Our results suggest a complex regulation of elastin gene expression during development, particularly within the proximal regions of the large arterial vessels.
Similar content being viewed by others
References
Albretsen C, Haukanes B-I, Aasland R, Kleppe K (1988) Optimal conditions for the hybridization with oligonucleotides: a study with myc-oncogene DNA probes. Anal Biochem 170:193–202
Anderson MLM, Young BD (1985) Quantitative filter hybridization In: Hames BD, Higgins SJ (eds) Nucleic acid hybridization. IRL Press, Oxford, England, pp 73–111
Badesch DB, Lee PDK, Parks WC, Stenmark KR (1989) Insulin-like growth factor I stimulates elastin synthesis by pulmonary arterial smooth muscle cells. Biochem Biophys Res Commun 160:382–387
Baule VJ, Foster JA (1988) Multiple chick tropoelastin mRNAs. Biochem Biophys Res Commun 154:1054–1060
Beck F, Samani NJ, Penschow JD, Thorley B, Tregear GW, Coghlan JP (1987) Histochemical localization of IGF-I and -II mRNA in the developing rat enbryo. Development 101:175–184
Bressan GM, Argos P, Stanley KK (1987) Repeating structure of chick tropoelastin revealed by complementary DNA. Biochemistry 26:1497–1503
Bruce MC (1991) Developmental changes in tropoelastin mRNA levels in rat lung: evaluation by in situ hybridization. Am J Respir Cell Mol Biol 5:344–350
Burnett W, Eichner R, Rosenbloom J (1980) Correlation of functional elastin messenger ribonucleic acid levels and rate of elastin synthesis in the developing chick aorta. Biochemistry 19:1106–1111
Burnett W, Finnigan-Bunick A, Yoon K, Rosenbloom J (1982) Analysis of elastin gene expression in the developing chick aorta using cloned elastin cDNA. J Biol Chem 257:1569–1572
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Damiano V, Tsang A-L, Weinbaum G, Christner P, Rosenbloom J (1984) Secretion of elastin in the embryonic chick aorta as visualized by immunoelectron microscopy. Coll Relat Res 4:153–164
Davidson JM, Shibahara S, Boyd C, Mason ML, Tolstoshev P, Crystal RG (1984) Elastin mRNA levels during foetal development of sheep nuchal ligament and lung. Biochem J 220:653–663
Davidson JM, Hill KE, Mason ML, Giro MG (1985) Longitudinal gradients of collagen and elastin gene expression in the porcine aorta. J Biol Chem 260:1901–1908
Deak SB, Pierce RA, Belsky SA, Riley DJ, Boyd CD (1988) Rat tropoelastin is synthesized from a 3.5-kilobase mRNA. J Biol Chem 263:504–507
Ernfors P, Henschen A, Olson L, Persson H (1989) Expression of nerve growth factor receptor mRNA is developmentally regulated and increased after axotomy in rat spinal cord motoneurons. Neuron 2:1605–1613
Fazio MJ, David RO, Eunkyung AK, Clinton TB, Indik Z, Ornstein-Goldstein N, Yeh H, Rosenbloom J, Uitto J (1988) Cloning of full length elastin cDNAs from a human skin fibroblast recombinant cDNA library: further elucidation of alternative splicing utilizing exon-specific oligonucleotides. J Invest Dermatol 91:458–464
Foster J, Rich CB, Florini JR (1987) Insulin-like growth factor I, somatomedin C, induces the synthesis of tropoelastin in aortic tissue. Coll Relat Res 7:161–169
Hamburger V, Hamilton LH (1951) A series of normal stages in the development of the chick embryo. J Morphol 88:49–92
Lathe R (1990) Oligonucleotide probes for in situ hybridization In: Polak JM, McGee JO'D (eds) In situ hybridization: principles and practice. Oxford University Press, New York, pp 71–80
Le Liévre CS, Le Douarin NM (1975) Mesenchymal derivatives of the neural crest: analysis of chimeric quail and chick embryos. J Embryol Exp Morphol 34:125–154
Noguchi A, Samaha H (1991) Developmental changes in tropoelastin gene expression in the rat lung studied by in situ hybridization. Am J Respir Cell Mol Biol 5:571–578
Parks WC, Secrist H, Wu LC, Mecham RP (1988) Developmental regulation of tropoelastin isoforms. J Biol Chem 263:4416–4423
Parks WC, Roby JD, Wu LC, Grosso LE (1992) Cellular expression of tropoelastin mRNA splice variants. Matrix 12:156–162
Pierce RA, Alatawi A, Deak SB, Boyd CD (1992) Elements of the rat tropoelastin gene associated with alternative splicing. Genomics 12:651–658
Presser IW, Stenmark KR, Suthar M, Crouch EC, Mecham RP, Parks WC (1989) Regional heterogeneity of elastin and collagen gene expression in intralobar arteries in response to hypoxic pulmonary hypertension as demonstrated by in situ hybridization. Am J Pathol 135:1073–1087
Rosenquist TH, Beall AC (1990) Elastogenesis in the developing cardiovascular system. Smooth muscle, nonmuscle, and cardiac neural crest. Ann N Y Acad Sci 588:106–119
Rosenquist TH, McCoy JR, Waldo KL, Kirby ML (1988) Origin and propagation of elastogenesis in the developing cardiovascular system. Anat Rec 221:860–871
Rosenquist TH, Fray-Gavalas C, Waldo K, Beall AC (1990) Development of the musculoelastic septation complex in the avian truncus arteriosus. Am J Anat 189:339–356
Selmin O, Volpin D, Bressan GM (1991) Changes of cellular expression of mRNA for tropoelastin in the intraembryonic arterial vessels of developing chick revealed by in situ hybridization. Matrix 11:347–358
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, USA, pp 743–752
Schmitz GG, Walter T, Seibl R, Kessler C (1991) Nonradioactive labeling of oligonucleotides with the hapten digoxigenin by tailing with terminal transferase. Anal Biochem 192:222–231
Sutcliffe MC, Davidson JM (1990) Effect of stretching on elastin production by porcine aortic smooth muscle cells. Matrix 10:48–153
Tokimitsu I, Tajima S, Nishikawa T, Ajima M, Fukasawa T (1987) Sequence analysis of elastin cDNA from chick aorta and tissue-specific transcription of the elastin gene in developing chick embryo. Arch Biochem Biophys 256:455–461
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Holzenberger, M., Liévre, C.AL. & Robert, L. Tropoelastin gene expression in the developing vascular system of the chicken: an in situ hybridization study. Anat Embryol 188, 481–492 (1993). https://doi.org/10.1007/BF00190142
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00190142