bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of tectal lesion on the survival of isthmic neurones in Xenopus
Authors: Straznicky Charles , Mccart Roger
Published: April 2021

The isthmic nucleus (IN) is a visual relay centre of the frog brain. It receives afferent projection from the optic tectum of the same side and projects bilaterally to both tecta. In young postmetamorphic Xenopus frogs, the survival of neurones in the IN on both sides was studied following the complete removal of the right tectum.

In 6- to 8-week-old frogs, the right tectum was surgically removed and the operated animals allowed to survive for 1 to 13 weeks after operation. In selected animals, 3 days before the intended sacrifice, the postoptic commissure was transected and the cut isthmotectal fibres filled with horseradish peroxidase (HRP). In serial paraffin sections of the midbrain, the numbers of surviving and dying (pyknotic) neurones in the left and right IN were counted. The soma size of viable isthmic neurones and the volume of both IN were measured.

Pyknotic neurones were seen between 1 and 6 weeks after operation in both the left and right IN, although the rate of cell loss was much greater in the latter.

Virtually all the neurones of the right IN degenerated by 6 weeks after tectal ablation. In contrast, approximately 60 % of neurones of the left IN survived. HRP histochemistry showed labelled isthmic neurones both in the left and right IN up to 3 weeks after operation. Thereafter, HRP-labelled neurones appeared only in the left IN.

These observations indicate that the removal of the natural target of isthmic neurones brings about severe neurone death. If target removal is accompanied by deafferentation, as is the case in the right IN, isthmic neurone death is exacerbated and all neurones die within a short period of 6 weeks. The 60 % neurone survival in the left IN suggests that a very substantial proportion of isthmic neurones projects initially, or is channelled as a result of tectal lesion, to the ipsilateral tectum.
Ordering of Y-specific sequences by deletion mapping and analysis of X-Y interchange males and females
Authors: Ferguson-Smith M. A. , Affara N. A. , Magenis R. E.
Published: April 2021

We have used DNA from 23 patients with Y-chromosome aberrations and 25 patients with presumptive X-Y interchange to map 39 Yp restriction fragments and 37 Yq restriction fragments. In the majority of patients the results are consistent with a standard contiguous order of sequences along the Y chromosome. In 6 of 26 patients (23 %) with Yp aberrations and 2 of 17 (12 %) with Yq aberrations, exceptions to the consensus order have been observed. These can be accommodated by postulating the presence of inversion polymorphisms. Such variation may occur more commonly on the nonpairing part of the Y chromosome that in other chromosomes owing to the absence of homologous synapsis and recombination in male meiosis.

The Y sequence most frequently present in X-Y interchange males was that recognized by GMGY3. 18 of 19 X-Y interchange males had this sequence suggesting that it is the nearest in the series to the TDF locus, and indicating that the latter maps to the distal end of Yp. Several techniques, including in situ hybridization and DNA measurement by flow cytometry, have been used to demonstrate that in X-Y interchange males there is transfer of Y sequences to the distal end of the X chromosome; no mechanism other than X-Y interchange has been demonstrated.
Primary sex determination in the nematode C. elegans
Authors: Hodgkin Jonathan
Published: April 2021

Most nematodes have XO male/XX female sex determination. C. elegans is anomalous, having XX hermaphrodites rather than females. The hermaphrodite condition appears to result from the modification of a basic male/female sex-determination system, which permits both spermatogenesis and oogenesis to occur within a female soma. This modification is achieved by a germ-line-specific control acting at one step in a cascade of autosomal regulatory genes, which respond to X-chromosome dosage and direct male, female, or hermaphrodite development. Mutations of one of these genes can be used to construct artificial strains with ZZ male/WZ female sex determination.

Primary sex determination normally depends on the ratio of X chromosomes to autosomes, as in Drosophila, and there appear to be multiple sites on the X chromosome that contribute to this ratio. Also, as in Drosophila, X-chromosome expression is compensated to equalize gene activity in XX and XO animals. Interactions between dosage compensation and sex determination are described and discussed.
Polytene chromosomes in mouse trophoblast giant cells
Authors: Varmuza Susannah , Prideaux Valerie , Kothary Rashmi , Rossant Janet
Published: April 2021

Mouse trophoblast giant cells undergo successive rounds of DNA replication resulting in amplification of the genome. It has been difficult to determine whether giant cell chromosomes are polyploid as in liver cells or polytene as in Dipteran salivary glands because the chromosomes do not condense. We have examined the pattern of hybridization of mouse giant cells with a variety of in situ chromosome markers to address this question. Hemizygous markers displayed one hybridization signal per nucleus in both diploid and giant cells, while homozygous markers displayed two signals per nucleus in both cell types. These patterns are consistent with cytological evidence indicating that giant cell chromosomes are polytene rather than polyploid. However, in contrast to the situation in Dipteran salivary glands, the two homologues do not appear to be closely associated. We conclude that the mechanism of giant cell DNA amplification involves multiple rounds of DNA replication in the absence of both karyokinesis and cytokinesis, and that sister chromatids, but not homologous chromosomes, remain closely associated during this process.
Graded requirement for the zygotic terminal gene,tailless, in the brain and tail region of theDrosophilaembryo
Authors: Strecker Teresa R. , Merriam John R. , Lengyel Judith A.
Published: April 2021

We have used hypomorphic and null tailless (tll) alleles to carry out a detailed analysis of the effects of the lack of tll gene activity on anterior and posterior regions of the embryo. The arrangement of tll alleles into a continuous series clarifies the relationship between the anterior and posterior functions of the tll gene and indicates that there is a graded sensitivity of anterior and posterior structures to a decrease in tll gene activity. With the deletion of both anterior and posterior pattern domains in tll null embryos, there is a poleward expansion of the remaining pattern. Using anti-horseradish peroxidase staining, we show that the formation of the embryonic brain requires tll. A phenotypic and genetic study of other pattern mutants places the tll gene within the hierarchy of maternal and zygotic genes required for the formation of the normal body pattern. Analysis of mutants doubly deficient in tll and maternal terminal genes is consistent with the idea that these genes act together in a common pathway to establish the domains at opposite ends of the embryo. We propose that tll establishes anterior and posterior subdomains (acron and tail regions, respectively) within the larger pattern regions affected by the maternal terminal genes.
Mouse homeobox gene transcripts occupy different but overlapping domains in embryonic germ layers and organs: a comparison of Hox-3.1 and Hox-1.5
Authors: Gaunt Stephen J.
Published: April 2021

By use of in situ hybridization experiments, the mouse homeobox genes Hox-3.1 and Hox-1.5 are compared in the temporal and spatial patterns of their embryonic transcripts. Transcripts of both genes are first detected at about days, although the appearance of Hox-3.1 transcripts apparently follows Hox-1.5 after a small delay. Hox-3.1 and Hox-1.5 transcripts occupy domains which are different, although overlapping, along the anteroposterior axis of the embryo. The domains are first established within the ectoderm and mesoderm germ layers at days, but subsequently they persist within the nervous system, the prevertebral column and within at least some of the organs (the thyroid, lung, stomach, mesonephric and metanephric kidneys) at days. In discussion, two different mechanisms are considered by which positional information might first be generated within the germ layers.
Regional expression, pattern and timing of convergence and extension during gastrulation ofXenopus laevis
Authors: Keller Ray , Danilchik Mike
Published: April 2021

We show with time-lapse micrography that narrowing in the circumblastoporal dimension (convergence) and lengthening in the animal-vegetal dimension (extension) of the involuting marginal zone (IMZ) and the noninvoluting marginal zone (NIMZ) are the major tissue movements driving blastopore closure and involution of the IMZ during gastrulation in the South African clawed frog, Xenopus laevis. Analysis of blastopore closure shows that the degree of convergence is uniform from dorsal to ventral sides, whereas the degree of extension is greater on the dorsal side of the gastrula. Expiants of the gastrula show simultaneous convergence and extension in the dorsal IMZ and NIMZ. In both regions, convergence and extension are most pronounced at their common boundary, and decrease in both animal and vegetal directions. Convergent extension is autonomous to the IMZ and begins at stage 10·5, after the IMZ has involuted. In contrast, expression of convergent extension in the NIMZ appears to be dependent on basal contact with chordamesoderm or with itself. The degree of extension decreases progressively in lateral and ventral sectors. Isolated ventral sectors show convergence without a corresponding degree of extension, perhaps reflecting the transient convergence and thickening that occurs in this region of the intact embryo. We present a detailed mechanism of how these processes are integrated with others to produce gastrulation. The significance of the regional expression of convergence and extension in Xenopus is discussed and compared to gastrulation in other amphibians.
Development and characterization of commissural interneurones in the spinal cord of Xenopus laevis embryos revealed by antibodies to glycine
Authors: Roberts Alan , Dale N. , Ottersen O. P. , Storm-Mathisen J.
Published: April 2021

By using an antibody to glutaraldehyde fixation products of glycine we have been able to observe the development of a defined population of spinal interneurones in the CNS of Xenopus laevis embryos. The first glycine immunoreactive (GLY) somata appeared at stage 22 in the caudal hindbrain within a few hours of neural tube closure. The population then increased by extending caudally into the spinal cord and by infill. It was followed up to the time of hatching, stage 37/38. By observing GLY cells at early stages in their differentiation, the normal sequence of cell process formation was deduced. A ventral axon is formed, extends dendrites laterally into the marginal zone and forms a commissure by growing through the ventral ependymal cell floor of the neural tube. On the opposite side, growth cones turn longitudinally and TEM observations show that they make en-passant synaptic contacts. All GLY cells have decussating axons and some grow secondary axons on the same side as the soma.

To establish the identity of GLY cells, a detailed comparison was made with commissural and dorsolateral commissural interneurones defined by retrograde and intracellular HRP staining. The GLY cells are identified with the commissural interneurones which are known to serve a glycinergic reciprocal inhibitory function. By showing that these interneurones have a clearly defined group identity and programme of development, this study opens the way to further experiments on factors controlling spinal cord pathway determination.
Characterization of retinoid metabolism in the developing chick limb bud
Authors: Thaller Christina , Eichele Gregor
Published: April 2021

Retinoids (vitamin A derivatives) have been shown to have striking effects on developing and regenerating vertebrate limbs. In the developing chick limb, retinoic acid is a candidate morphogen that may coordinate the pattern of cellular differentiation along the anteroposterior limb axis. We describe a series of investigations of the metabolic pathway of retinoids in the chick limb bud system. To study retinoid metabolism in the bud, all-trans-[3H]retinol, all-trans-[3H]retinal and all-trans-[3H]retinoic acid were released into the posterior region of the limb anlage, the area that contains the zone of polarizing activity, a tissue possibly involved in limb pattern formation. We found that the locally applied [3H]retinol is primarily converted to [3H]retinal, [3H]retinoic acid and a yet unidentified metabolite. When [3H]retinal is locally applied, it is either oxidized to [3H]retinoic acid or reduced to [3H]retinol. In contrast, local delivery of retinoic acid to the bud yields neither retinal norretinol nor the unknown metabolite. This flow of metabolites agrees with the biochemical pathway of retinoids that has previously been elucidated in a number of other animal systems. To find out whether metabolism takes place directly in the treated limb bud, we have compared the amount of [3H]retinoid present in each of the four limb anlagen following local treatment of the right wing bud. The data suggest that retinoid metabolism takes place mostly in the treated limb bud. This local metabolism could provide a simple mechanism to generate in a controlled fashion the biologically active all-trans-retinoic acid from its abundant biosynthetic precursor retinol. In addition, local metabolism supports the hypothesis that retinoids are local chemical mediators involved in pattern formation.
The control of DNA replication in a cell-free extract that recapitulates a basic cell cycle in vitro
Authors: Hutchison Christopher J. , Cox Richard , Ford Christopher C.
Published: April 2021

Cell-free extracts prepared from Xenopus eggs support chromosome decondensation and pronuclear formation on demembranated sperm heads. 32P-dCTP pulse-labelling studies demonstrate that DNA synthesis occurs in multiple bursts of 30–40 min in extracts containing pronuclei, each burst being followed by a period of 20–50 min during which no synthesis occurs. Density substitution with bromodeoxyuridine indicates that the synthesis in each burst is semiconservative and results from new initiations, and that, following multiple bursts of synthesis, reinitiation events can occur. Changes in nuclear morphology have been characterized in the extract by phase-contrast microscopy and by fluorescence microscopy following pulse labelling with biotin-11-dUTP and staining with anti-lamin antibodies. Lamin accumulation occurs as DNA decondenses and parallels the acquisition of membrane structures. Biotin-11-dUTP incorporation is first observed in small nuclei having decondensed DNA and an extensive lamina. While DNA synthesis is occurring nuclei remain relatively small, but rapid swelling accompanied by chromosome condensation occurs when biotin incorporation ceases. Nuclear swelling and chromatin condensation is followed by nuclear membrane breakdown, lamin dispersal and chromosome formation. Mitosis lasts for approximately 20 min. Nuclear reassembly is recognized by the appearance of membrane vesicles around small pieces of decondensed DNA, which parallels the appearance of lamin islands within a chromatin mass. These ‘islands’ incorporate biotin, indicating that DNA synthesis is occurring, and apparently fuse as larger S-phase nuclei are formed. Extensive protein synthesis occurs for at least 4h in most extracts. This synthesis is required for the initiation of mitotic events and the reinitiation of DNA synthesis.
Behaviour of microtubules and actin filaments in livingDrosophilaembryos
Authors: Kellogg Douglas R. , Mitchison Tim J. , Alberts Bruce M.
Published: April 2021

We describe the preparation of novel fluorescent derivatives of rabbit muscle actin and bovine tubulin, and the use of these derivatives to study the behaviour of actin filaments and microtubules in living Drosophila embryos, in which the nuclei divide at intervals of 8 to 21 min. The fluorescently labelled proteins appear to function normally in vitro and in vivo, and they allow continuous observation of the cytoskeleton in living embryos without perturbing development. By coinjecting labelled actin and tubulin into the early syncytial embryo, the spatial relationships between the distinct filament networks that they form can be followed second by second. The dynamic rearrangements of actin filaments and microtubules observed confirms and extends results obtained from previous studies, in which fixation techniques and specific staining were used to visualize the cytoskeleton in the Drosophila embryo. However, no tested fixation method produces an exact representation of the in vivo microtubule distribution.
DNA methylation in the developing marsupial embryo
Authors: Stevens Mary E. , Maidens Peta M. , Robinson Edward S. , Vandeberg John L. , Pedersen Roger A. , Monk Marilyn
Published: April 2021

Marsupial development differs from early development of placental mammals in that the blastocyst is unilaminar, so that both embryonic and extraembryonic cells are derived from a single layer of cells (protoderm) which faces the blastocyst cavity. Also, all cells in female marsupial conceptuses so far examined show preferential paternal X-inactivation. To test for a possible correlation between cell position, paternal X-inactivation and DNA hypomethylation, marsupial DNA preparations from three regions, embryo, vascular yolk sac and avascular yolk sac, were digested with methyl-specific restriction endonucleases, separated on agarose gels and end-labelled with 32P-dCTP. The size distribution of the fragments obtained indicated three levels of methylation: high methylation of embryonic DNA, intermediate levels of methylation of vascular yolk sac DNA and hypomethylation of avascular yolk sac DNA. The degree of methylation of repeat sequences, observed as discrete bands in end-labelled HpaII digests, was correlated with the overall methylation of tissue DNA. Thus, the difference in methylation in embryonic and extraembryonic DNA was similar to that described for the mouse conceptus, and the outside cell position of marsupial fetal precursor cells did not correlate with hypomethylation. HpaII tiny fragments, which indicate the presence of CpG-rich islands of DNA, were evident in the marsupial digests. In the mouse DNA, these islands are associated with gene transcription and provide one route to cloning of unique gene sequences.
Development of the pattern of cell renewal in the crypt-villus unit of chimaeric mouse small intestine
Authors: Schmidt Günter H. , Winton Douglas J. , Ponder Bruce A. J.
Published: April 2021

We have previously shown that the epithelium of each adult intestinal crypt in chimaeric mice is derived from a single progenitor cell. Whether the crypts are monoclonal from the outset - that is, are formed by the proliferation of a single cell - or whether their formation is initiated by several cells was not known. Here we report that many crypts contain cells of both chimaeric genotypes in the neonatal period indicating a polyclonal origin at this stage of morphogenesis. The cellular organization of the early neonatal crypt is therefore different from that of the adult crypt, which includes a zone of ‘anchored’ stem cells above the crypt base. Within 2 weeks, however, the crypt progenitor cell and its descendants displace all other cells from the crypt and the crypt attains monocle-nality. The distribution of enterocytes on chimaeric villi in the neonate shows a mottled pattern of mosaicism which is progressively replaced by coherent sheets of cells from the crypts, and within two weeks the orderly adult clonal pattern is established.
Stalk cell formation in monolayers from isolated prestalk and prespore cells of Dictyostelium discoideum: evidence for two populations of prestalk cells
Authors: Kwong Linda , Sobolewski André , Atkinson Leone , Weeks Gerald
Published: April 2021

Cells from the pseudoplasmodial stage of Dictyostelium discoideum differentiation were dispersed and separated on Percoll gradients into prestalk and prespore cells. The requirements for stalk cell formation in low-density monolayers from the two cell types were determined. The isolated prespore cells required both the Differentiation Inducing Factor (DIF) and cyclic AMP for stalk cell formation. In contrast, only part of the isolated prestalk cell population required both cyclic AMP and DIF, the remainder requiring DIF alone, suggesting the possibility that there were two populations of prestalk cells, one independent of cyclic AMP and one dependent on cyclic AMP for stalk cell formation. The finding that part of the prestalk cell population required only a brief incubation in the presence of DIF to induce stalk cell formation, whilst the remainder required a considerably longer incubation in the presence of both DIF and cyclic AMP was consistent with this idea. In addition, stalk cell formation from cyclic-AMP-dependent prestalk cells was relatively more sensitive to caffeine inhibition than stalk cell formation from cyclic-AMPindependent prestalk cells. The latter cells were enriched in the most anterior portion of the migrating pseudoplasmodium, indicating that there is spatial segregation of the two prestalk cell populations.

The conversion of prespore cells to stalk cells took longer and was more sensitive to caffeine when compared to stalk cell formation from cyclic-AMP-dependent prestalk cells.
Two maternally derived X chromosomes contribute to parthenogenetic inviability
Authors: Mann Jeff R. , Lovell-Badge Robin H.
Published: April 2021

In certain extraembryonic tissues of normal female mouse conceptuses, X-chromosome-dosage compensation is achieved by preferential inactivation of the paternally derived X. Diploid parthenogenones have two maternally derived X chromosomes, hence this mechanism cannot operate. To examine whether this contributes to the inviability of parthenogenones, XO and XX parthenogenetic eggs were constructed by pronuclear transplantation and their development assessed after transfer to pseudopregnant recipients. In one series of experiments, the frequency of post implantation development of XO parthenogenones was much higher than that of their XX counterparts. This result is consistent with the possibility that two maternally derived X chromosomes can contribute to parthenogenetic inviability at or very soon after implantation. However, both XO and XX parthenogenones showed similar developmental abnormalities at the postimplantation stage, demonstrating that parthenogenetic inviability is ultimately determined by the possession of two sets of maternally derived autosomes.
Requirements for X-linked zygotic gene activity during cellularization of early Drosophila embryos
Authors: Wieschaus Eric , Sweeton Dari
Published: April 2021

To examine the requirements for X-chromosomal transcription during precellular stages of Drosophila embryogenesis, attached X-chromosomes and XY translocations were used to generate embryos deficient for large cytologically defined regions of that chromosome. Embryos that lack all X-chromosome material (‘nullo-X embryos’) develop normally to the cycle-14 syncytial blastoderm stage, but fail to partition their nuclei to single cells during cellularization. The cellularization defects can first be detected in the abnormal distribution of cortical actin and nuclei during early cycle 14. The same defects are produced by deletions of only a single region on the X-chromosome, between 6F and 7A. Nullo-X embryos carrying a duplication of this region cellularize and develop normally to the onset of gastrulation.
The localization of an inductive response
Authors: Gurdon J. B.
Published: April 2021

Combinations of tissues from Xenopus blastulae have been used to identify several mechanisms that limit the number of animal cells forming muscle after induction by vegetal cells. The results disagree with a model in which direct physical contact or very close proximity between animal and vegetal cells restricts the number of cells that receive the inductive signal. Rather it seems that a diffusible inducer is released by vegetal cells, and spreads through 4–8 animal cell diameters, equivalent to a distance of 80 μm, from the nearest vegetal cells. Several factors seem to cooperate to prevent the further spread of the mesoderm-forming induction. These include the slow diffusion and/or instability of the inducer, the time of loss of competence of animal cells to respond to induction, and the amount of vegetal tissue that releases inducer for a limited time. The combination of these, and perhaps other, processes seems to ensure that a consistent minority of animal cells are induced to form muscle, thereby leaving other animal cells available to form the nervous system and epidermis.
Blastocyst implantation precedes induction of insulin-like growth factor II gene expression in human trophoblasts
Authors: Ohlsson Rolf , Larsson Erik , Nilsson Ove , Wahlström Torsten , Sundström Per
Published: April 2021

The cytotrophoblast cell population of the human embryonic conceptus proliferates rapidly during the first month following blastocyst implantation. Since the trophectoderm lineage is established in preimplantation morula/blastocysts, the scenario underlying initiation and maintenance of the rapid proliferative phenotype of cytotrophoblasts is a central issue. The insulin-like growth factor II (IGF-II) gene is highly expressed in proliferative cytotrophoblasts of first trimester placenta and performs as a placenta growth factor. To establish a temporal correlation between IGF-II expression and initiation of highly proliferative trophoblasts in human development, we employed in situ hybridization analysis of the expression of the IGF-II and human chorionic gonadotropin β-subunit (β-HCG) genes in human pre- and postimplantation development. The data show that the appearance of high steady-state levels of IGF-II transcripts in trophoblasts is a postimplantation event, whereas β-HCG transcripts can already be detected in preimplantation development. This observation makes a role for endogenously produced IGF-II in the normal development of preimplantation embryos unlikely, but suggests that endogenously produced IGF-II participates in the formation and subsequent expansion of the rapid proliferative phenotype of the trophoblastic shell, following implantation.
Complex regulation of TGFβ expression by retinoic acid in the vitamin A-deficient rat
Authors: Glick Adam B. , Mccune Bryan K. , Abdulkarem Nariman , Flanders Kathleen C. , Lumadue Jeanne A. , Smith Joseph M. , Sporn Michael B.
Published: April 2021

We report the results of a histochemical study, using polyclonal antipeptide antibodies to the different TGFβ isoforms, which demonstrates that retinoic acid regulates the expression of TGFβ in the vitamin A-deficient rat. Basal expression of TGFβ diminished under conditions of vitamin A deficiency. Treatment with retinoic acid caused a rapid and transient induction of TGFβ2 and TGFβ3 in the epidermis, tracheobronchial and alveolar epithelium, and intestinal mucosa. Induction of TGFβ1 expression was also observed in the epidermis. In contrast to these epithelia, expression of the three TGFβ isoforms increased in vaginal epithelium during vitamin A deficiency, and decreased following systemic administration of retinoic acid. Our results show for the first time the widespread regulation of TGFβ expression by retinoic acid in vivo, and suggest a possible mechanism by which retinoids regulate the functions of both normal and pre-neoplastic epithelia.
Interaction of zona pellucida glycoproteins, sulphated carbohydrates and synthetic polymers with proacrosin, the putative egg-binding protein from mammalian spermatozoa
Authors: Jones Roy
Published: April 2021

Fertilization in mammals is a unique cell-cell recognition event that involves specific receptors on the surface of each gamete. Previous work has shown that proacrosin, a protein found within the acrosome of mammalian spermatozoa, binds non-enzymatically to zona pellucida glycoproteins (ZPGPs) that surround the egg and that this binding can be inhibited by sulphated polysaccharides such as fucoidan. The mechanism of this interaction has been investigated using 125I-ZPGPs and 125I-fucoidan as probes. Results show that it involves poly(sulphate) groups on zona glycoproteins that bind with high affinity (Kd=1.2 to 5.0×10−8M) to complementary ‘docking’ sites on proacrosin. The spatial orientation of these sulphates, together with the tertiary structure of the target protein, determines the selectivity of polymer binding. Thus, dextran sulphate and poly(vinyl sulphate) are strong inhibitors of the above probes whereas dextran, chondroitin sulphates A and C and poly(vinyl phosphate) are ineffective. Proacrosin, therefore, has properties analogous to those described for ‘bindin’, the egg adhesion protein found within the acrosomal vesicle of sea urchin spermatozoa.
Localization of TIMP in cycling mouse hair
Authors: Kawabe Thomas T. , Rea Thomas J. , Flenniken Ann M. , Williams B. R. G. , E. Groppi Vincent , Buhl Allen E.
Published: April 2021

TIMP (tissue inhibitor of metalloproteinase) is a glycoprotein inhibitor of metalloproteinases that we hypothesize to be involved in the tissue remodeling that occurs during each hair growth cycle. We examined this hypothesis by studying the expression of TIMP at selected times during a single hair cycle using T1MP– lacZ transgenic mice to localize TIMP gene activity in the hair follicle. TIMP gene induction was visualized by staining mouse back skin for β-galactosidase (β-gal) activity. Paraffin sections were analyzed for the localization of TIMP expression. TIMP gene activation appears in hair follicles only during the mid-anagen (the growing stage of the hair cycle) primarily in Henle’s layer of the inner root sheath. Some expression of TIMP is also seen in a few connective tissue cells, in the sebaceous gland and in cells at the proximity of the dermal papilla cells in catagen (regressing) and telogen (resting) follicles. These results are consistent with a role for TIMP in cyclic remodeling of connective tissue in hair follicles.
The restricted spatial and temporal expression of a nervous-system-specific antigen involved in axon outgrowth during development of the grasshopper
Authors: Seaver Elaine C. , Karlstrom Rolf O. , Bastiani Michael J.
Published: April 2021

To identify molecules important for pathfinding by growing axons, monoclonal antibodies (mAb) have been generated against embryonic grasshopper tissue. One mAb, 2B2, shows labeling exclusively in the nervous system. It recognizes a surface epitope on neuronal growth cones, filopodia and axons in the central nervous system (CNS). Initially, the antigen is expressed on all processes of the CNS; after 70% of embryonic development, localization of the 2B2 mAb is restricted to a small subset of axon tracts within the ganglia. Immunoprecipitation from embryonic membrane extracts with the 2B2 mAb reveals a unique band of 160×103Mr. Functional studies with the 2B2 mAb demonstrate that the antigen is important in growth cone-axon interactions during process outgrowth. Growth cones that extend along axonal substrata are either blocked in growth or grow along an aberrant pathway when embryos are cultured in the presence of the 2B2 mAb. However, pioneer neurons that extend processes on non-neuronal substrata grow normally.
Control of cell fate in the tail region of C. elegans by the gene egl-5
Authors: Chisholm Andrew
Published: April 2021

The tail region of C. elegans contains a number of blast cell and neuron types that either are found only in the tail, or are different from more anterior homologues. In egl-5 mutants, the fates of many of these tail cells are abnormal or transformed to those of anterior homologues. The affected cells are related only by position and not by ancestry, egl-5 is also required for normal development of the somatic gonad and sex muscles in males. The function of egl-5 in all these tissues is cell autonomous. By genetic mapping, egl-5 lies very close to mab-5, a gene with an analogous role in the immediately anterior body region, egl-5 and mab-5 may constitute a ‘mini-cluster’ of regional determination genes, similar to those described in other animal phyla.
Spatial mechanisms of gene regulation in metazoan embryos
Authors: Davidson Eric H.
Published: April 2021

The basic characteristics of embryonic process throughout Metazoa are considered with focus on those aspects that provide insight into how cell specification occurs in the initial stages of development. There appear to be three major types of embryogenesis: Type 1, a general form characteristic of most invertebrate taxa of today, in which lineage plays an important role in the spatial organization of the early embryo, and cell specification occurs in situ, by both autonomous and conditional mechanisms; Type 2, the vertebrate form of embryogenesis, which proceeds by mechanisms that are essentially independent of cell lineage, in which diffusible morphogens and extensive early cell migration are particularly important; Type 3, the form exemplified by long germ band insects in which several different regulatory mechanisms are used to generate precise patterns of nuclear gene expression prior to cellularization. Evolutionary implications of the phylogenetic distribution of these types of embryogenesis are considered. Regionally expressed homeodomain regulators are utilized in all three types of embryo, in similar ways in later and postembryonic development, but in different ways in early embryonic development. A specific downstream molecular function for this class of regulator is proposed, based on evidence obtained in vertebrate systems. This provides a route by which to approach the comparative regulatory strategies underlying the three major types of embryogenesis.
LIF, the ES-cell inhibition factor, reversibly blocks nephrogenesis in cultured mouse kidney rudiments
Authors: Bard Jonathan B. L. , Ross Allyson S. A.
Published: April 2021

Mouse kidney induction proceeds in vitro much as it does in vivo: the ureteric hud bifurcates to give collecting ducts while the mesenchyme condenses into aggregates which epithelialise and then elongate into tuhules with glomerular and other nephron structures. We report here that the factor known as LIF (leukaemia inhibitory factor), which regulates the differentiation and growth of embryonic-stem (ES) and other cells in culture, has little effect in vitro on growth or on ureteric-bud morphogenesis other than to stimulate the bifurcation process. It does however exert a striking effect on the mesenchyme. At about four times the concentration required to inhibit ES-cell differentiation, LIF strongly but reversibly blocks the effects of metanephric mesenchyme induction: although mesenchyme condenses around growing duct tips, the number of mature nephrons that form over 6 days is reduced by 75 % or more. The few nephrons that do develop in the presence of LIF probably come from mesenchyme already induced at the time of culture and are indistinguishable from those that form in controls as assayed by morphology, by X-gal staining of endogenous galactosidase and by antibodies to brush-border and CD15 antigens. There is a further unexpected feature of rudiments cultured in LIF which is absent in controls: they contain an unexpectedly high number of stable epithelialised aggregates that express laminin around their periphery and which do not develop further. These results argue that the process of nephrogenesis involves at least two distinct stages which can be blocked by LIF: the effect of the initial induction and the future development of epithelialised aggregates. The ability of LIF to regulate the differentiation of kidney blast cells as well as a diverse range of other mammalian cell types thus implies that there are common events associated with entering new developmental pathways.
A homolog of Drosophila Notch expressed during mammalian development
Authors: Weinmaster Gerry , Roberts Veronica J. , Lemke Greg
Published: April 2021

Drosophila Notch and the related Caenorhabditis elegans proteins lin-12 and glp-1 function as mediators of local cell-cell interactions required for cell-fate decisions during invertebrate development. To investigate the possibility that similar proteins play determinative roles during mammalian development, we isolated cDNA clones encoding rat Notch. The deduced amino acid sequence of this protein contains 36 epidermal growth factor (EGF)-like repeats, and is remarkably similar in both its extracellular and cytoplasmic domains to the sequence of Xenopus Xotch and Drosophila Notch. In the developing central nervous system, in situ hybridisation analyses revealed that Notch transcripts were dramatically restricted to the ventricular proliferative zones of embryonic neuroepithelia. Notch was also strongly expressed during development of non-neural tissues, such as hair follicles and tooth buds, whose correct differentiation requires epithelial-mesenchymal interactions. These data support the hypothesis that Notch plays an essential role in mammalian development and pattern formation that closely parallels its role in the development of invertebrates.
Transcription factor AP-2 is tissue-specific in Xenopus and is closely related or identical to Keratin Transcription Factor 1 (KTF-1)
Authors: Snape Alison M. , Winning Robert S. , Sargent Thomas D.
Published: April 2021

This paper identifies a new, developmental role for transcription factor AP-2 in the activation of amphibian embryonic epidermal keratin gene expression. Keratin transcription factor KTF-1 is shown by several criteria to be identical or closely related to AP-2. KTF-1/AP-2 is shown to be tissue-specific from its first transcription in Xenopus embryos, and restricted to a small number of adult tissues, including skin. Epidermis-specific keratin transcription closely follows specification of the embryonic ectoderm in Xenopus, and is subject to regulation by growth factors and embryonic induction. We further show that in mouse basal kératinocytes, a KTF-l/AP-2-like factor is present and binds to a DNA sequence previously shown to be important in the regulation of the keratin K14 gene, which is actively expressed in these cells. Thus, the study of AP-2 and its role in the regulation of keratin gene transcription should enhance our understanding of both amphibian embryonic development and mammalian skin differentiation.
Tissue-specific control of expression of the tight junction polypeptide ZO-1 in the mouse early embryo
Authors: Fleming Tom P. , Hay Mark J.
Published: April 2021

The processes governing differential protein expression in preimplantation lineages were investigated using a monoclonal antibody recognising the tight junction polypeptide, ZO-1. ZO-1 localises to the maturing tight junction membrane domain in the polarised trophectoderm lineage from compaction (8-cell stage) onwards, ultimately forming a zonular belt around each trophectoderm cell of the blastocyst (32- to 64-cell stage). The protein is usually undetectable within the inner cell mass (ICM) although, in a minority of embryos, punctate ZO-1 sites are present on the surface of one or more ICM cells. Since ICM cells derive from the differentiative division of polarised 8- and 16-cell blastomeres, the distribution of ZO-1 following differentiative division in isolated, synchronised cell clusters of varying size, was examined. In contrast to the apical cytocortical pole, ZO-1 was found to be inherited by nonpolar (prospective ICM) as well as polar (prospective trophectoderm) daughter cells. Following division, polar cells adhere to and gradually envelop nonpolar cells. Prior to envelopment, ZO-1 localises to the boundary between the contact area and free membrane of daughter cells, irrespective of their phenotype. After envelopment, polar cells retain these ZO-1 contact sites whilst nonpolar cells lose them, in which case ZO-1 transiently appears as randomly-distributed punctate sites on the membrane before disappearing. Thus, symmetrical cell contact appears to initiate ZO-1 down-regulation in the ICM lineage. The biosynthetic level at which ZO-1 down-regulation occurs was investigated in immunosurgically isolated ICMs undergoing trophectoderm regeneration. By 6 h in culture, isolated ICMs generated a zonular network of ZO-1 at the contact area between outer cells, thereby demonstrating the reversibility of down-regulation. This assembly process was unaffected by alpha-amanitin treatment but was inhibited by cycloheximide. These results indicate that the ICM inherits and stabilises ZO-1 transcripts which can be utilised for rapid synthesis and assembly of the protein, a capacity that may have significance both in maintaining lineage integrity within the blastocyst and in the subsequent development of the ICM.
Expression of the cystic fibrosis gene in human development
Authors: Harris Ann , Chalkley Gillian , Goodman Shelley , Coleman Lindsay
Published: April 2021

The specialised epithelia lining the respiratory tract, pancreatic ducts, male genital ducts and sweat gland ducts are defective in the severe inherited disease, cystic fibrosis (CF).

We have looked at the expression of the CF gene in human fetal tissues to throw light on the development of function in specialised ductal epithelia and to determine the age of onset of the CF disease process. The CF gene is already seen to be transcribed in mid-trimester fetal lung, pancreas and male genital ducts. Hence, by this developmental stage, and before they are fully differentiated, these epithelia have the capability to perform important transport functions. Epithelial cell cultures derived from fetal pancreas and male genital ducts maintain expression of the CF gene in vitro and so form good models for analysing CF gene function and differentiation of these specialised epithelia.
Down-regulation of the chicken α5β1 integrin fibronectin receptor during development
Authors: Muschler John L. , Horwitz Alan F.
Published: April 2021

We have characterized the diversity of the chicken β1 integrin family and studied the expression of individual receptors during development. The diversity of the β1 integrin family was investigated by affinity purifying the β1 integrins from a variety of adult and embryonic tissues. These purifications reveal the relative levels of expression and also the differential expression of the α subunits in those tissues. Monoclonal antibodies were generated against the prominent ‘band 1’ of the embryonic chicken integrins and used to characterize the expression of this α subunit in embryonic and adult tissues. This α subunit is shown to be the chicken homologue of human α5 fibronectin receptor. The chicken α5β1 integrin is the most prominent β1 integrin in the embryo and is expressed on the majority of cell types through the day 17 stage. The distribution of this receptor in the embryo closely parallels the distribution of its ligand, fibronectin. In adult tissues, expression of this receptor is greatly diminished relative to the expression of other α subunits. The cell type distribution is highly restricted: limited primarily to the vasculature and to connective tissue regions. These studies reveal a prominent role for the α5β1 integrin in embryonic cell types and a down-regulation of this receptor on many cell types during development.
The control of cell fate along the dorsal-ventral axis of the Drosophila embryo
Authors: Ray Robert P. , Arora Kavita , Nüsslein-Volhard Christiane , Gelbart William M.
Published: April 2021

We have analyzed the contributions made by maternal and zygotic genes to the establishment of the expression patterns of four zygotic patterning genes: decapenta- plegic (dpp), zerkniillt (zen), twist (twi), and snail (sna). All of these genes are initially expressed either dorsally or ventrally in the segmented region of the embryo, and at the poles. In the segmented region of the embryo, correct expression of these genes depends on cues from the maternal morphogen dorsal (dl). The dl gradient appears to be interpreted on three levels: dorsal cells express dpp and zen, but not twi and sna; lateral cells lack expression of all four genes; ventral cells express twi and sna, but not dpp and zen. dl appears to activate the expression of twi and sna and repress the expression of dpp and zen. Polar expression of dpp and zen requires the terminal system to override the repression by dl, while that of twi and sna requires the terminal system to augment activation by dl. The zygotic expression patterns established by the maternal genes appear to specify autonomous domains that carry out independent developmental programs, insofar as mutations in the genes that are expressed ventrally do not affect the initiation or ontogeny of the expression patterns of the genes that are expressed dorsally, and vice versa. However, interactions between the zygotic genes specific to a particular morphological domain appear to be important for further elaboration of the three levels specified by dl. Two of the genes, dpp and twi, are unaffected by mutations in any of the tested zygotic dorsal-ventral genes, suggesting that dpp and twi are the primary patterning genes for dorsal ectoderm and mesoderm, respectively.
Expression of p53 during mouse embryogenesis
Authors: Schmid Peter , Lorenz Andreas , Hameister Horst , Montenarh Mathias
Published: April 2021

By in situ hybridisation we have examined the expression of p53 during mouse embryogenesis from day 8.5 to day 18.5 post coitum (p.c.). High levels of p53 mRNA were detected in all cells of the day 8.5 p.c. and 10.5 p.c. mouse embryo. However, at later stages of development, expression became more pronounced during differentiation of specific tissues e.g. of the brain, liver, lung, thymus, intestine, salivary gland and kidney. In cells undergoing terminal differentiation, the level of p53 mRNA declined strongly. In the brain, hybridisa-tion signals were also observed in postmitotic but not yet terminally differentiated cells. Therefore, gene ex-pression of p53 does not appear to be linked with cellular proliferation in this organ. A proposed role for p53 in cellular differentiation is discussed.