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Field homology: a meaningful definition.

Field homology refers to populations of cells that derive from evolutionarily conserved regions of embryos but are distributed across sets of adult morphological structures that cannot be placed in one-to-one correspondance. The concept of field homology has proven especially attractive to comparative neurologists because it allows them to deal with the fact that sets of nuclei or nuclear subdivisions often cannot be compared on a one-to-one basis across phyletic groups. However, the concept of field homology has recently come under criticism. It has been argued that field homology is theoretically impossible because it requires sequences of developmental stages to be both evolutionarily conserved and evolutionarily modified. It has also been argued that field homology allows overly vague comparisons of adult morphological structures, fails to account for homologous structures that derive from non-homologous embryonic sources, and establishes overly rigid links between embryonic and adult morphology. All of these criticisms may be adequately addressed by explaining field homology in terms of differentiation. The present paper explains field homology in terms of differentiation using the amniote dorsal thalamus to illustrate major points. It is concluded that field homology is a meaningful concept when defined in terms of differentiation, applied to appropriate cases, and properly limited in its comparisons of adult structures.

Animals↗

SUMOylation of Tr2 orphan receptor involves Pml and fine-tunes Oct4 expression in stem cells.

The Tr2 orphan nuclear receptor can be SUMOylated, resulting in the replacement of coregulators recruited to the regulatory region of its endogenous target gene, Oct4. UnSUMOylated Tr2 activates Oct4, enhancing embryonal carcinoma-cell proliferation, and is localized to the promyelocytic leukemia (Pml) nuclear bodies. When its abundance is elevated, Tr2 is SUMOylated at Lys238 and seems to be released from the nuclear bodies to act as a repressor. SUMOylation of Tr2 induces an exchange of its coregulators: corepressor Rip140 replaces coactivator Pcaf, which switches Tr2 from an activator to a repressor. This involves dynamic partitioning of Tr2 into Pml-containing and Pml-free pools. These results support a model where SUMOylation-dependent partitioning and differential coregulator recruitment contribute to the maintenance of a homeostatic supply of activating, as opposed to repressive, Tr2, thus fine-tuning Oct4 expression and regulating stem-cell proliferation.

Animals↗

Primary structure of the long and short splice variants of mouse collagen XII and their tissue-specific expression during embryonic development.

Type XII collagen, a member of the FACIT group of extracellular matrix proteins, consists of molecules that are trimers of alpha 1(XII) chains. The three chains in each molecule form a cross-shaped structure with a central globule from which a triple-helical tail and three finger-like regions (containing von Willebrand factor A-like regions (containing von Willebrand factor A-like domains and fibronectin type III repeats) extend. cDNA cloning/sequencing of chicken alpha 1(XII) collagen and protein studies with mouse, bovine, and human material suggest that the alpha 1(XII) collagen gene gives rise to two molecular variants, differing in the length of the finger-like regions, by alternative splicing of the primary transcript. To provide a basis for studies of the function of the two variants in an organism that can be genetically manipulated, we have isolated and sequenced mouse cDNAs encoding both splice variants. The sequence provides the first complete nucleotide and amino acid sequence of mammalian type XII collagen. From these cDNAs we have generated digoxigenin-labeled RNA probes for in situ hybridization of developing mouse embryos to find out whether the splicing mechanism responsible for generation of the two forms is developmentally regulated. The results, combined with Northern blot and RT-PCR analysis of RNA from embryos at various developmental stages, demonstrate that the long form of collagen XII, XIIA, is the predominant form at early stages (ED7 and 11); at later stages of development (ED15 and 17) the short form, XIIB, becomes the major form. As the short form becomes the major product, the long splice variant continues to be expressed in several tissues, even after birth. An exception is dermis, which is positive for the long form up to embryonic day 15, but negative at day 18, when only the short form RNA can be detected.

Alternative Splicing↗

A truncated activin receptor inhibits mesoderm induction and formation of axial structures in Xenopus embryos.

Activins can induce mesoderm in embryonic explants and have been proposed as the natural inducer in Xenopus. A mutant activin receptor that inhibits activin signalling is used to show that activin is required for the induction of mesoderm in vivo and the patterning of the embryonic body plan. Blocking the activin signal transduction pathway also reveals autonomous induction of a neural marker and unmasks a relationship between activin and fibroblast growth factor.

Activin Receptors↗

[Structural changes in the mitotic apparatus of metaphase cells exposed to 2-mercaptoethanol in a porcine embryonic kidney cell culture. A light microscopy study].

The sequence of reconstruction of the metaphase mitotic apparatus of the KEPV cells under the action of 2-mercaptoethanol (0.001 M) and after moving off the agent has been studied. In the presence of 2-mercaptoethanol, a destruction of the metaphase plate and that of the mitotic spindle proceed independently. The destruction of the mitotic spindle does not involve an immediate disorganization of the metaphase plate, and vice versa. In the course of resumption of the normal division after moving off the agent, the restoration of the mitotic apparatus structure proceeds within a definite temporal sequence: at first, the mitotic spindle reestablishes, and after that the metaphase plate forms. Distinctions between the 2-mercaptoethanol action and the action of other antimitotic agents are discussed.

Animals↗

Structure-function studies of anti-3-fucosyllactosamine (Le(x)) and galactosylgloboside antibodies.

We are studying murine mAbs against two carbohydrate epitopes, 3-fucosyllactosamine (Le(x), CD15) and galactosylgloboside. The VH domains of both panels of Ab are encoded by VH441, a member of the X24 family of Ig genes. To evaluate the contribution of the heavy chain CDR3 to the affinity of the anti-3-fucosyllactosamine Ab, CDR3-H of PMN6, a low affinity Ab, was replaced by the CDR3 of PM81, a higher affinity Ab. The affinity of the chimeric 6/81 Ab was increased when the heavy chain was paired with the PM81 light chain, but not when paired with another light chain (M5), which differs from PM81 light chain by three amino acids. To evaluate the contribution of somatic mutations to the binding of GalGb4, the 3A9 VH sequence, which contains three amino acid substitutions, was replaced by a germ-line sequence encoded by either VH441 or VHX24. The chimeric Ab, 441/3A9 and X24/3A9, bound Ag as well as the wild-type 3A9 Ab. Computer models of the Fv fragments of PM81 and 3A9 were compared with the crystal structure of the Fv fragment of J539, a galactan-binding myeloma protein that is encoded by the same VH and VK genes as 3A9. The surfaces of 3A9 and J539 have shallow pockets that are potential Ag-binding sites. Replacement of CDR3-H Tyr99, which is a prominent component of the pocket, by Ala abolished the binding of Ag. In contrast, the Fv surface of PM81 contains a large cleft rather than a pocket. These models indicate how the same VH gene segment can be used to encode Abs that exhibit different specificities.

Animals↗

Structure of a cDNA for Ciona Cytochrome b(5) and the ubiquitous expression of mRNA in embryonic tissues.

A cDNA clone for cytochrome b(5) was isolated from a cDNA library of an ascidian, Ciona savignyi, by a plaque hybridization method using a digoxigenin-labeled cDNA for the soluble form of human cytochrome b(5). The cDNA is composed of 5'- and 3'-noncoding sequences, and a 396-base pair coding sequence. The 3'-noncoding sequence contains polyadenylation signal sequences. The amino acid sequence of 132 residues deduced from the nucleotide sequence of the cDNA showed 61% identity and 82% similarity to the cytochrome b(5) of another ascidian species, Polyandrocarpa misakiensis, which we previously cloned. The amino-terminal hydrophilic domain of 98 residues contains well-conserved structures around two histidine residues for heme binding. A cDNA expression system was constructed to prepare a putative soluble form of Ciona cytochrome b(5). The recombinant soluble cytochrome b(5) showed an asymmetrical absorption spectrum at 560 nm as is shown by mammalian cytochromes b(5) upon reduction with NADH and NADH-cytochrome b(5) reductase. The recombinant Ciona cytochrome b(5) is reduced by NADH-cytochrome b(5) reductase with an apparent K(m) value of 3.3 microM. This value is similar to that of the cytochrome b(5) of Polyandrocarpa misakiensis. The expression of Ciona cytochrome b(5) mRNA during development was examined by an in situ hybridization method and ubiquitous expression in embryonic tissues was observed. The results indicate that cytochrome b(5) plays important roles in various metabolic processes during development.

Amino Acid Sequence↗

The temporary anatomical structures prominent in the first trimester may be fulfilling exchange functions assigned to the placenta in the second and third trimester.

The extra-embryonic coelom (EEC) and secondary yolk sac are prominent structures in the gestational sac during the first trimester of human pregnancy, at a time before the definitive placental circulation becomes established. We propose that the EEC and yolk sac play a critical role in the nutrition of early pregnancy, fulfilling exchange functions which are assumed by the placenta at a later stage.

Amniotic Fluid↗

Effects of gonadal sex and incubation temperature on the ontogeny of gonadal steroid concentrations and secondary sex structures in leopard geckos, Eublepharis macularius.

Incubation temperature during embryonic development determines gonadal sex in the leopard gecko (Eublepharis macularius). Incubation temperature and gonadal sex jointly influence the display of sexual and agonistic behavior in adult leopard geckos. These differences in adult behavior are organized prior to sexual maturity, and it is plausible that post-natal hormones influence neural and behavioral differentiation. Here we assessed incubation temperature and sex effects on sex steroid levels in leopard geckos at 2, 10, and 25 weeks of age and monitored the development of male secondary sex structures. Males had significantly higher androgen concentrations at all time points, whereas females had significantly higher 17beta-estradiol (E2) concentrations only at 10 and 25 weeks. Within males, age but not incubation temperature affected steroid levels and morphological development. Male androgen levels increased modestly by 10 and dramatically by 25 weeks of age, whereas E2 levels remained unchanged over this period. Most males had signs of hemipenes at 10 weeks of age, and all males had hemipenes and open preanal pores by 25 weeks of age. In females, age and incubation temperature affected E2 and dihydrotestosterone (DHT) but not T concentrations. Controlling for age, females from 34 degrees C have higher DHT and lower E2 levels than females from 30 degrees C. Further, E2 concentrations increased significantly from 2 to 10 weeks, after which E2 levels remained steady. Together, these results indicate that sexually dimorphic levels of steroids play a major role in the development of leopard gecko behavior and morphology. Furthermore, these data suggest that the organizational effects of incubation temperature on adult female phenotype could be, in part, mediated by incubation temperature effects on steroid hormone levels during juvenile development.

Age Factors↗

Ultrastructural aspects of the production of extracellular matrix components by the chick embryonic notochord in vitro.

The morphology of extracellular matrix (ECM) components and of the cell organelles, particularly the Golgi complex and its derived structures, implicated in the production of ECM in the chick embryonic notochord have been studied by transmission electron microscopy. Isolated notochordal fragments were cultured in suspension in liquid medium. Native striated collagen fibrils with a period of 540 A were observed in the perinotochordal sheath. Fine granular and filamentous materials suggestive of proteoglycans have been observed in intercellular spaces and under the basal lamina of the notochordal sheath. Golgi mature vesicles with structures resembling the previously described segment-long-spacing (SLS)-like aggregates and secretory vesicles probably containing proteoglycans or condensed collagen precursors have also been observed.

Animals↗

The matrix Gla protein gene is a marker of the chondrogenesis cell lineage during mouse development.

Matrix Gla protein (MGP) is, along with osteocalcin, a skeletal member of the family of extracellular mineral-binding Gla proteins. Although the precise function of these proteins remains obscure, circumstantial evidence suggests that they play a role in endochondral ossification. As a first step toward understanding MGP function we have performed a preliminary characterization of its promoter element and studied the developmental pattern of expression of this gene. DNA transfection experiments indicate that the mouse MGP promoter functions better in cells expressing the MGP gene than in cells that do not express the gene. During mouse development, MGP gene expression is detectable as early as day 10.5 of embryonic development (E10.5), before any skeletal structures are identifiable. In situ hybridization analysis shows that MGP mRNA is initially present at the mesenchymal epithelial interphase in lung and limb buds. As development proceeds, MGP gene is predominantly expressed in cells of the chondrocytic lineage in areas that will undergo endochondral ossification as well as in areas that will remain cartilaginous, such as the trachea and bronchi. In growth plate cartilage, MGP mRNA is present in resting, proliferative, and late hypertrophic chondrocytes. Surprisingly, MGP mRNA is absent from the early hypertrophic chondrocytes and from the osteoblasts. Finally, the MGP gene is expressed at a lower level in kidney medulla and uterus smooth muscle but not in brain, spleen, or heart during development. This study demonstrates that during development MGP gene expression occurs early and is predominant at the epithelial mesenchymal interfaces, principally of lung and limb buds, and in cells of the chondrocytic lineage. This finding raises the intriguing possibility that MGP may play distinct roles during embryogenesis and in the adult organism.

Animals↗

Contribution of neurons born during embryonic, juvenile, and adult life to the brain of adult canaries: regional specificity and delayed birth of neurons in the song-control nuclei.

Neurogenesis occurs in adult song birds, which suggests that neurons born after hatching may contribute to histogenesis and plasticity of the avian brain. However, little is known about the overall contribution to the mature brain of neurons born in juveniles and adults, and how this process affects different regions of the avian brain. In fact, studies of the histogenesis of the avian forebrain have made the classical assumption that neuronal birth ends before hatching. Here we determined the contribution of neurons born before and after hatching to different regions throughout the adult canary brain. Male canaries were injected with [3H]-thymidine at different times during embryonic, juvenile, and adult life. The position of labeled neurons was mapped in parasagittal brain sections. Because all birds were killed as adults, results indicate the time of birth of neurons that survived to adulthood in different structures of the avian brain. Injection at embryonic day (E) 5 or E9 resulted in labeled neurons in all regions of the neuroaxis. The vast majority of neurons outside of the telencephalon were born before E9. One exception was a discrete region in the dorsal thalamus, a part of the song-control circuit, where neurons continued to be born after E9. Most regions of the telencephalon had a high proportion of its neurons labeled by the embryonic injections. In particular, archistriatum, anterior neostriatum, and the hippocampus had most of their neurons labeled before hatching. This indicates that many of the telencephalic neurons born in the embryo are long lived and are not replaced by other neurons that continue to be added to the telencephalon after hatching. Neurons labeled by [3H]-thymidine injections after hatching were restricted to the telencephalon and contributed importantly to many regions. In particular, the avian striatum (lobus parolfactorius, LPO) received a large number of its neurons during the first 20 days of life, but continued to incorporate new neurons throughout juvenile and adult life. Neurons continued to be added to the telencephalon of adults (even in 4-year-old birds). The distribution of labeled neurons after [3H]-thymidine injections in adults was similar to that observed in latter stages of juvenile development. The contribution of neurons born at different ages from embryonic development to adulthood varied among different anatomical subdivisions of the canary brain. this could, in part, explain differences in the cytoarchitecture and plasticity between brain regions. Neurogenesis after hatching may allow the modification of selected brain circuits as the bird matures and ages.

Animals↗

Histone deacetylase activity is required for embryonic stem cell differentiation.

Mammalian development requires commitment of cells to restricted lineages, which requires epigenetic regulation of chromatin structure. Epigenetic modifications were examined during in vitro differentiation of murine embryonic stem (ES) cells. Global histone acetylation, a euchromatin marker, declines dramatically within 1 day of differentiation induction and partially rebounds by day 2. Histone H3-Lys9 methylation, a heterochromatin marker, increases during in vitro differentiation. Conversely, the euchromatin marker H3-Lys4 methylation transiently decreases, then increases to undifferentiated levels by day 4, and decreases by day 6. Global cytosine methylation, another heterochromatin marker, increases slightly during ES cell differentiation. Chromatin structure of the Oct4 and Brachyury gene promoters is modulated in concert with their pattern of expression during ES cell differentiation. Importantly, prevention of global histone deacetylation by treatment with trichostatin A prevents ES cell differentiation. Hence, ES cells undergo functionally important global and gene-specific remodeling of chromatin structure during in vitro differentiation. genesis 38:32-38, 2004.

Acetyltransferases↗

Fine structural distribution of acetylcholine receptors at developing mouse neuromuscular junctions.

The distribution of acetylcholine receptors (AChRs) in neuromuscular junctions of embryonic and newborn mice (strain 129/ReJ) was examined on the fine structural level using [125I]alpha-bungarotoxin labeling and quantitative electron microscope autoradiography. Changes in junctional receptor site density were related to changes in the structure of the postjunctional membrane, in particular the differentiation of morphologically distinct regions of thickened membrane and the formation of junctional folds. The following sequence of development is described: (1) At the earliest age examined, embryonic day 16 of gestation (i.e., at approximately the time when junctional receptor accumulations are first detected), subneural receptor aggregates have poorly defined boundaries and seem to extend beyond the region of direct axonal or Schwann cell contact. At that time the subneural AChR site density is somewhat variable but averages approximately 2000 to 3000 sites/micron 2, and the subneural muscle membrane shows discontinuous membrane specialization (membrane thickening plus a cytoplasmic amorphous layer). There seems to be no preferential labeling of contact regions rich in such specialization compared with those where membrane thickening was less obvious or absent. (2) By birth, junctional AChRs are strongly correlated with morphologically specialized membrane, at a constant density of approximately 9000/micron 2 (comparable to that in adult animals of this strain of mouse). During the first postnatal week, a large fraction of the primary cleft is covered by Schwann cell rather than axon terminal. Receptor-rich dense membrane and incipient junctional folds are found under axon terminals and, to a lesser extent, under Schwann cell extensions, but they are not seen beyond the edge of the "junctional complex." Coated vesicles are a prominent feature of the subjunctional sarcoplasm in neonatal junctions, and their number declines sharply during the first week. (3) Junctional folds differentiate mainly after the first postnatal week. Development of folds is associated with a 1.5- to 1.8-fold increase in length of specialized thickened membrane and a parallel increase in number of receptor sites per endplate. Since this is also a period of rapid overall endplate growth (Steinbach, J.H. (1981) Dev. Biol. 84: 267-276) the total number of endplate receptors increases greater than 30-fold during the maturation period following the first week while the concentration of receptors/micron 2 of thickened membrane remains constant.

Animals↗

Altered podocyte structure in GLEPP1 (Ptpro)-deficient mice associated with hypertension and low glomerular filtration rate.

Glomerular epithelial protein 1 (GLEPP1) is a receptor tyrosine phosphatase present on the apical cell surface of the glomerular podocyte. The GLEPP1 gene (PTPRO:) was disrupted at an exon coding for the NH(2)-terminal region by gene targeting in embryonic stem cells. Heterozygote mating produced the expected genotypic ratio of 1:2:1, indicating that the Ptpro(-/-) genotype does not lead to embryonic or neonatal lethality. Kidney and glomerular structure was normal at the gross and light microscopic levels. Scanning and transmission electron microscopy showed that Ptpro(-/-) mice had an amoeboid rather than the typical octopoid structure seen in the wild-type mouse podocyte and that there were blunting and widening of the minor (foot) processes in association with altered distribution of the podocyte intermediate cytoskeletal protein vimentin. Reduced filtration surface area in association with these structural changes was confirmed by finding reduced glomerular nephrin content and reduced glomerular filtration rate in Ptpro(-/-) mice. There was no detectable increase in the urine albumin excretion of Ptpro(-/-) mice. After removal of one or more kidneys, Ptpro(-/-) mice had higher blood pressure than did their wild-type littermates. These data support the conclusion that the GLEPP1 (Ptpro) receptor plays a role in regulating the glomerular pressure/filtration rate relationship through an effect on podocyte structure and function.

Albumins↗

Cytokine regulation of secondary lymphoid organ development.

Lymphotoxin and tumor necrosis factor provide essential signals for the formation of secondary lymphoid tissue structures. Lymphotoxin in its membrane form (LT alpha 1 beta 2 heterotrimer) is required for the development of lymph nodes and Peyer's patches and supports the development of normal spleen structure. In the spleen, lymphotoxin acts during embryonic development to support the formation of distinct B and T cell zones. Lymphotoxin also acts in a tonic fashion-supporting the formation and maintenance of the follicular dendritic cell network and of primary B cell follicle structure. The cells that deliver the tonic lymphotoxin signal supporting follicular dendritic cell structure are B cells; thus, B cells participate fundamentally in the development of the lymphoid tissue structure in which they subsequently mature.

Animals↗

[Histochemistry and electron microscopy of the embryonal development of the golden hamster kidney: study of the interstitial cells].

The structure and the origin of the interstitial cells in the kidney of the golden Hamster during its embryonic development was studied. For this purpose kidneys from 11, 13, 15 and 17 day-old embryos were used. The histochemical observations as well as the electron microscopy demonstrated that the interstitial cells have a metanephric origin and are embryonically similar to the cellular elements that differentiate the primordial renal vesicles and later organise the epithelial portion of the nephrons. These cells seem to take part in the formation of the basal membranes during tubular genesis.

Acid Phosphatase↗

Positional cloning of a global regulator of anterior-posterior patterning in mice.

Anterior-posterior (A-P) patterning is of fundamental importance throughout vertebrate embryonic development. Murine members of the trithorax (trx) and Polycomb group (Pc-G) of genes regulate A-P patterning of segmented axial structures, demonstrating conserved upstream regulation of homeotic pathways between Drosophila and mouse. Here we report the positional cloning of a classical mouse mutation, eed (for embryonic ectoderm development), which is the highly conserved homologue of the Drosophila Pc-G gene esc (for extra sex combs), a long-term repressor of homeotic genes. Mutants homozygous for a null allele of eed display disrupted A-P patterning of the primitive streak during gastrulation. Mutant embryos lack a node, notochord and somites, and there is no neural induction. In contrast to absence of anterior structures, extra-embryonic mesoderm is abundant. Mice carrying a hypomorphic eed mutation exhibit posterior transformations along the axial skeleton. These results indicate that eed is required globally for A-P patterning throughout embryogenesis.

Amino Acid Sequence↗