Elsevier

Gene Expression Patterns

Volume 5, Issue 1, November 2004, Pages 75-79
Gene Expression Patterns

Isolation and embryonic expression of avian ADAM 12 and ADAM 19

https://doi.org/10.1016/j.modgep.2004.06.003Get rights and content

Abstract

Members of the ADAM gene family encode large multi-domain proteins containing A Disintegrin And Metalloprotease domain. We have cloned quail orthologs of ADAM 12 and 19 using PCR-based screening and describe their expression patterns over the period E2.5 (Hamilton and Hamburger stage 14) to E5.0 (HH 25) using in situ hybridisation. Quail ADAM 12 is expressed in mesenchyme, cranially, in the tail and in the limb buds, and also in visceral mesenchyme. In the nervous system it is expressed in dorsal root ganglia and ventral roots. Quail ADAM 19 is expressed in cranial and dorsal root ganglia, sympathetic ganglia, ventral mixed nerves and in the allantois. Avian ADAM 12 and 19 genes exhibit similarities and differences in expression pattern compared to their murine orthologs, for example, expression of ADAM 12 in the nervous system, limb and tail bud in quail but not mouse. Interestingly, in mouse ADAM 19 is expressed in these locations. We have generated a sheep antibody to quail ADAM 19 and, in embryonic cells in vitro, found the protein at cell–cell junctions in many cell types. Some of these did have detectable ADAM 19 by in situ hybridisation but RT-PCR analysis confirmed the presence of low level ADAM 19 transcripts not detectable by in situ hybridisation.

Section snippets

Results and discussion

The ADAM (A Disintegrin And Metalloprotease domain) genes encode large trans-membrane proteins with extracellular metalloprotease, disintegrin- and cysteine-rich domains and a putative intracellular signalling domain. The meltrin sub-family consists of ADAM 12, 13, 19 and 33. ADAM 13 is characterised only in Xenopus, and is expressed in cranial neural crest cells migrating to the branchial arches, and in somitic mesoderm (Alfandari et al., 1997). ADAM 13 perturbation experiments disturb cranial

Materials and methods

Fertile Japanese quail eggs (Coturnix coturnix japonica) (Lago Smallgoods, Melbourne, Australia) were incubated at 37 °C and staged according to Hamburger and Hamilton (1951) for chick embryos. Note that the morphological stage series (denoted HH) of quails is essentially identical to that of chicks, but the chronology of development, denoted by embryonic (E) day, is slightly more rapid.

A quail cDNA fragment with homology to the meltrin class ADAMs was initially obtained by two rounds of

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