The BMP antagonists cerberus-like and noggin do not interact during mouse forebrain development.

Mouse cerberus-like encodes for a secreted factor of the Cerberus/Dan family. This molecule has neural inducing capabilities and can bind to BMP-4 and nodal molecules in the extracellular space. When cerberus-like is inactivated, its function may be compensated for another molecule, since no abnormalities can be observed in the mouse mutant. Compensation mechanisms have been shown to occur between the BMP antagonists chordin and noggin. Here we report the generation of cerberus-like-/-; noggin-/- double mutants to uncover a possible compensation by noggin in cer-l-/- mutant. Double mutants were obtained and failed to show any further detectable defects beside the ones presented by the noggin-/- single mutant. Contrarily to chordin and noggin, mouse cerberus-like and noggin cannot compensate for each other during mouse embryogenesis.

Cerberus-like is a secreted BMP and nodal antagonist not essential for mouse development.

Mouse cerberus-like (cer-l) is a member of the Cerberus/Dan family of secreted factors. As other members of this family of proteins, Cer-l functions in the extracellular space, inhibiting signaling molecules. Here we show that the neural-inducing and mesoderm-inhibiting activities of Cer-l result from specific binding to BMP and Nodal molecules, respectively. These properties resemble the ones from the related factor Xenopus Cerberus. However, Xenopus Cerberus in addition to BMP4 and Nodal also binds to and inhibits Wnt proteins. We show that Cer-l does not directly inhibit Wnt signals. A null allele of the mouse Cer-l gene was generated by targeted inactivation in ES cells. Homozygous embryos show no anterior patterning defects, are born alive, and are fertile. Since mouse Cer-l and Xenopus Cerberus differ in biochemical activities, we propose the existence of additional members of this family of inhibitors, which may compensate for the loss of cer-l.

The organizer factors Chordin and Noggin are required for mouse forebrain development.

In mice, there is evidence suggesting that the development of head and trunk structures is organized by distinctly separated cell populations. The head organizer is located in the anterior visceral endoderm (AVE) and the trunk organizer in the node and anterior primitive streak. In amphibians, Spemann's organizer, which is homologous to the node, partially overlaps with anterior endoderm cells expressing homologues of the AVE markers cerberus, Hex and Hesx1. For mice, this raises the question of whether the AVE and node are independent of each other, as suggested by their anatomical separation, or functionally interdependent as is the case in amphibians. Chordin and Noggin are secreted bone morphogenetic protein (BMP) antagonists expressed in the mouse node, but not in the AVE. Here we show that mice double-homozygous mutants that are for chordin and noggin display severe defects in the development of the prosencephalon. The results show that BMP antagonists in the node and its derivatives are required for head development.

Goosecoid regulates the neural inducing strength of the mouse node.

The homeobox gene goosecoid was the first specific genetic marker of Spemann's organizer in vertebrate embryos to be discovered. In the frog, misexpression of this gene by RNA injection produces duplication of the posterior axis. For these reasons, the recent finding that mice lacking goosecoid function have no early axial defects was rather surprising. Here we assay the neural inducing strength of wild-type and goosecoid-mutant mouse nodes by transplantation into primitive streak stage chick embryos. Wild-type mouse nodes strongly induce the neural-specific transcription factors Sox2 and Sox3 in the chick host. Homozygous goosecoid(-/- )nodes are severely impaired in their ability to induce both genes. Heterozygous goosecoid(+/-) nodes induce Sox3 as well as wild-type nodes, but resemble -/- nodes in their limited ability to induce Sox2. We propose that goosecoid does play a role in regulating the neural inducing strength of the node and that regulative mechanisms exist which mask the early phenotypic consequences of goosecoid mutations in the intact mouse embryo.

The prechordal midline of the chondrocranium is defective in Goosecoid-1 mouse mutants.

Gsc-1 expression marks cells with Spemann organizer, or axis-inducing, activity in the vertebrate gastrula. Gsc-1 knockouts, however, did not display phenotypes related to the early phase of expression. In this paper, additional phenotypes for the Gsc-1 mouse mutant are presented. Examination of the base of the cranium in the dorsal view revealed fusions and deletions in the midline of the prechordal chondrocranium. These defects were correlated with the sites of expression of Gsc-1 in the prechordal plate/foregut endoderm in the day 7.5/8.5 embryo. Gsc-1 expression in proximal limb buds was correlated with malformations of the shoulder and hip articulations. In addition, ribs in the seventh cervical vertebra were observed with low penetrance. The role of Gsc-1 during gastrulation and axial development is discussed in relation to possible compensatory interactions with other genes such as HNF-3beta and the recently identified Gsc-2 and Gsc-3 genes.

Cerberus-like is a secreted factor with neutralizing activity expressed in the anterior primitive endoderm of the mouse gastrula.

We report the isolation of mouse cerberus-like (cer-l), a gene encoding a novel secreted protein that is specifically expressed in the anterior visceral endoderm during early gastrulation. Expression in the primitive endoderm starts before the appearance of the primitive streak and lasts until the head-fold stage. In later stages, a second region of expression is found in newly formed somites. Mouse cer-l shares some sequence similarity with Xenopus cerberus (Xcer). In Xenopus assays cer-l, like Xcer, mRNA acts as a potent neuralizing factor that induces forebrain markers and endoderm, but is unable to induce ectopic head-like structures as Xcer does. In addition to cer-l, anterior visceral endoderm was found to express the transcription factors Lim1, goosecoid and HNF-3beta that are also present in trunk organizer cells. A model of how head and trunk development might be regulated is discussed. Given its neuralizing activity, the secreted protein Cer-l is a candidate for mediating inductive activities of anterior visceral endoderm.

Complementary DNA sequences of the 24 kDa and 21 kDa subunits of complex I from Neurospora.

We have cloned and sequenced cDNAs coding for two subunits of the peripheral arm of Neurospora crassa complex I. The two polypeptides are synthesized as precursor proteins which are processed to mature forms with predicted molecular masses of 24331 and 20982 Da.

Primary structure of the nuclear-encoded 10.5 kDa subunit of complex I from Neurospora crassa.

We have isolated a cDNA clone for the nuclear encoded 10.5 kDa subunit of complex I from N. crassa. DNA sequencing revealed an open reading frame corresponding to a polypeptide with 94 amino acids and a calculated molecular mass of 10531 Da. The protein is synthesized without a cleavable mitochondrial targeting sequence. The N. crassa polypeptide is the fungal equivalent of subunit B8 of bovine complex I.