bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for an association between U1 RNA and interspersed repeat single-copy RNAs in the cytoplasm of sea urchin eggs†
Authors: Ruzdijic Sabera , Pederson Thoru
Published: April 2021

Psoralen crosslinking of RNA–RNA intermolecular duplexes in sea urchin egg extracts reveals that some maternal poly(A)+ RNA molecules are complexed with U1 RNA, a cofactor in somatic nuclear pre-mRNA splicing. Reaction of egg extracts with a monoclonal antibody specific for U1 snRNP selects, in addition to Ul, RNAs that contain repeated sequences interspersed with single-copy elements. Antibodyselection experiments with nucleate and anucleate egg halves demonstrate that most of the Ul RNA–inter-spersed RNA complexes are cytoplasmic, as is the egg’s store of total Ul snRNP. These results raise the possibility that maternal interspersed RNAs include unprocessed pre-messenger RNA molecules in arrested complexes with splicing cofactors.
Gastrulation movements provide an early marker of mesoderm induction in Xenopus laevis
Authors: Symes K. , Smith J. C.
Published: April 2021

The first inductive interaction in amphibian development is mesoderm induction, in which an equatorial mesodermal rudiment is induced from the animal hemisphere under the influence of a signal from vegetal pole blastomeres. We have recently discovered that the Xenopus XTC cell line secretes a factor which has the properties we would expect of a mesoderm-inducing factor. In this paper, we show that an early response to this factor by isolated Xenopus animal pole regions is a change in shape, involving elongation and constriction. We show by several criteria, including general appearance, timing, rate of elongation and the nonrequirement for cell division that these movements resemble the events of gastrulation. We also demonstrate that the movements provide an early, simple and reliable indicator of mesoderm induction and are of use in providing a ‘model system’ for the study of mesoderm induction and gastrulation. For example, we show that the timing of gastrulation movements does not depend upon the time of receipt of a mesoderm-induction signal, but on an intrinsic gastrulation ‘clock’ which is present even in those animal pole cells that would not normally require it.
The postimplantation development of spontaneous digynic triploid embryos in LT/Sv strain mice
Authors: Kaufman M. H. , Speirs S.
Published: April 2021

When spontaneously ovulating LT/Sv female mice are mated with fertile males, between one third and one half of the zygotes analysed at the first cleavage mitosis are found to be triploid. This is due to the fact that LT/Sv females ovulate both primary and secondary oocytes, all of which are capable of being fertilized. Fertilization of the former group results in the production of digynic triploid conceptuses, while their diploid littermates result from the fertilization of normal secondary oocytes. The present study was therefore carried out in order to investigate the ‘spontaneous’ level of triploidy in these mice, and to provide insight into the developmental fate of the LT/Sv triploid embryos, as previous studies had indicated that in this species triploids invariably fail to develop beyond the early postimplantation period. This study revealed that when autopsies were carried out on the 7th and 8th days of gestation, it was generally difficult to distinguish between the kary-ologically normal diploids and the digynic triploid conceptuses when only morphological criteria were used. However, by the 10th day of gestation, the triploid conceptuses could usually be readily distinguished from their diploid littermates by their smaller size and (occasionally) by their disorganized or abnormal morphological appearance.
Modification of cell division by phorbol ester in preimplantation mouse embryos
Authors: Mystkowska Ewa T. , Sawicki Wojciech
Published: April 2021

2-cell mouse embryos were treated in vitro with a 2 h pulse of phorbol myristate acetate (PMA) at 32nd, 38th and 50th h after hCG, then chased in culture for up to 46 h. Embryos were fixed at various time intervals of chasing, then stained and inspected. Some embryos were carefully inspected with a video recording system, every l·44s and the cell divisions (cytokinesis) as well as formation of large, single blastomeres, each from two smaller ones, were recorded. PMA pulse led to the suppression of cell divisions. The rate of the suppression was time dependent: with a delay of 0–1, 12 and 18 h between the PMA pulse and time of scheduled cell division about 99, 87 and 44 % of 2-cell embryos remained at this stage of development, for at least 10 h, respectively, and 90, 58 and 12 % of their blastomeres revealed binu-clearity. Since we found that PMA-mediated formation of binuclearity was not the effect of cell fusions, it was assumed that the inhibition of cytokinesis preceded by karyokinesis was reponsible for binuclearity. PMA effect on cell divisions was reversible. PMA-treated embryos revealed formation of large, single blastomeres, each from two smaller ones. If cell division appeared after PMA pulse, in about 52 % of 3-to 6-cell embryos, the large blastomere formation was recorded in the course of the subsequent 38 h. Large blastomere formation was concluded to be the result of either cell fusion or reversion of incompleted cytokinesis brought about by PMA.
Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells*
Authors: Greenburg Gary , Hay Elizabeth D.
Published: April 2021

In considering the mechanism of transformation of epithelium to mesenchyme in the embryo, it is generally assumed that the ability to give rise to fibroblast-like cells is lost as epithelia mature. We reported previously that a definitive embryonic epithelium, that of the anterior lens, gives rise to freely migrating mesenchyme-like cells when suspended in type I collagen matrices. Here, we show that a highly differentiated epithelium that expresses cytokeratin changes to a vimentin cytoskeleton and loses thyroglobulin during epithelial-mesenchymal transformation induced by suspension in collagen gel. Using dispase and collagenase, we isolated adult thyroid follicles devoid of basal lamina and mesenchyme, and we suspended the follicles in 3D collagen gels. Cells bordering the follicle lumen retain epithelial polarity and thyroid phenotype, but basal cell surface organization is soon modified as a result of tissue multilayering and elongation of basal cells into the collagenous matrix. Cytodifferentiation, determined by thyroglobulin immunoreactivity, is lost as the basal epithelial cells move into the matrix after 3 – 4 days in collagen. By TEM, it can be seen that the elongating cells acquire pseudopodia, filopodia and mesenchyme-like nuclei and RER. Immunofluorescence examination of intermediate filaments showed that freshly isolated follicles and follicles cultured on planar substrata react only with anticytokeratin. However, all of the inesen-chyme-like cells express vimentin and they gradually lose cytokeratin. These results suggest that vimentin may be necessary for cell functions associated with migration within a 3D matrix. The mesenchymal cells do not revert to epithelium when grown on planar substrata and the transformation of epithelium to mesenchyme-like cells does not occur within basement membrane gels. The results are relevant to our understanding of the initiation of epithelial-mesenchymal transformation in the embryo and the genetic mechanisms controlling cell shape, polarity and cytoskeletal phenotype.
The early development of the atrioventricular node and bundle of His in the embryonic chick heart. An electrophysiological and morphological study
Authors: Argüello Carlos , Alanis Jesus , Valenzuela Blanca
Published: April 2021

The development of the atrioventricular node and bundle of His of embryonic chick hearts was studied by electrophysiological and morphological techniques. The dorsal wall of the AV canal and the interatrial septum were explored to determine if they contribute to the formation of the AV node and bundle of His. The resting membrane and action potentials of the interatrial septum cells were systematically analysed and found to undergo progressive differentiation with development. The earliest identification of the AV node and upper bundle of His group of cells was achieved at days of development by the electrical recording of their corresponding characteristic action potentials, from a circumscribed area located in the lowest and dorsal segment of the interatrial septum.

The morphological and anatomical characterization of the cells was made following electrical recording and labelling with charcoal particles. The earlier AV node and bundle of His responses had similar characteristics to those of the adult heart. It is concluded that the AV node and upper bundle of His cells derive from the low interatrial septum. The possibility that AV canal cells contribute to this event was discarded. The functional relationship of the AV node and bundle of His with other cardiac tissues during the early development of the heart is discussed.
Spatial organization of the epithelium and the role of neural crest cells in the initiation of the mammalian tooth germ
Authors: Lumsden A. G. S.
Published: April 2021

Teeth develop from composite organ rudiments that are formed through the interaction of oral epithelium and mesenchyme of the first branchial arch; cells of the former differentiate into enamelsecreting ameloblasts whereas those of the latter differentiate into dentinesecreting odontoblasts. Experimental analysis of odontogenic tissue interactions in mammalian embryos has focused on the late developmental stages of morphogenesis and cytodifferentiation; little is known about initial pattern-forming events, during which presumptive tooth-forming cells are specified and the sites of tooth initiation become established. It requires to be shown, for example, whether the mesenchymal cells of mammalian teeth are derived, like those of amphibians, from the cranial neural crest, and if so, whether these form a specified subpopulation in the neural folds. Alternatively, are they specified after migration into the mandibular arch, possibly by interaction with the oral epithelium?

The developmental potentials of mouse embryo premigratory cranial neural crest cells (CNC explanted from the caudal mesencephalic and rostral metencephalic neural folds) have been studied in intraocular homograft recombinations with various regions of embryonic surface ectoderm. Cartilage, bone and neural tissue developed in all combinations of CNC and epithelium. Teeth formed in combinations of CNC with mandibular arch epithelium but not in combinations of CNC with limb bud epithelium. Teeth also formed in combinations of mandibular arch epithelium with neural crest explanted from the trunk level. These results indicate that mammalian neural crest has an odontogenic potential but that this is not restricted to the crest of presumptive toothforming levels. Normal migration appears not to be a prerequisite for expression of odontogenic potential but this does require an interaction with regionspecific epithelium. It is reasonable to infer that during normal development the neural crest that enters the mandibular arch is odontogenically unspecified before or during migration and that the oral epithelium is the earliest known site of tooth pattern.
Palate development
Authors: Ferguson Mark W. J.
Published: April 2021

In all vertebrates, the secondary palate arises as bilateral outgrowths from the maxillary processes. In birds and most reptiles, these palatal shelves grow initially horizontally, but do not fuse with each other resulting in physiological cleft palate. In crocodilians, shelf fusion occurs resulting in an intact secondary palate. Mammalian palatal shelves initially grow vertically down the side of the tongue, but elevate at a precise time to a horizontal position above the dorsum of the tongue and fuse with each other to form an intact palate. Palatal shelf-elevation is the result of an intrinsic shelf elevating force, chiefly generated by the progressive accumulation and hydration of hyaluronic acid. In all vertebrates the nasal epithelium differentiates into pseudostratified ciliated columnar cells and the oral epithelia differentiates into stratified squamous cells, but the medial edge epithelial (MEE) phenotype differs in different groups. In mammals, the MEE of opposing shelves adhere to each other to form an epithelial seam which then disrupts by cell death and cell migration into the mesenchyme accompanied by an epitheliomesenchymal transformation. In birds, the MEE keratinize resulting in cleft palate whereas, in alligators, the MEE migrate onto the nasal aspect of the palate. In all vertebrates, this regional, temporal and species-specific epithelial differentiation is specified by the underlying mesen-chyme. Signalling of this interaction is complex but involves both extracellular matrix and soluble factors e.g. minor collagen types, tenascin, EGF, TGFα, TGFβ, PDGF, FGF. These soluble growth factors have a biphasic effect: directly on the epithelia and on the mesenchyme where they stimulate or inhibit cell division and synthesis of specific extracellular matrix molecules. The extracellular matrix molecules (and bound growth factors) synthesized by the mesenchymal cells may then directly affect the epithelium. These signals cause differential gene expression via second messenger systems e.g. cAMP, cGMP, Ca2+, pH, pl etc. Molecular markers for nasal, medial and oral epithelial cell differentiation include the types of cytokeratin intermediate filaments and specific cell surface molecules recognized by monoclonal antibodies: the genes for such molecules are probably expressed in response to mesenchymal signals. Using such an approach, it is possible to go from a morphological description of palate development to a cellular analysis of the mechanisms involved and then to identification of candidate genes that may be important for screening and diagnosis of cleft palate.
Cell-adhesion and substrate-adhesion molecules: their instructive roles in neural crest cell migration
Authors: Tucker Gordon C. , Duband Jean-Loup , Dufour Sylvie , Thiery Jean Paul
Published: April 2021

The individualization of crest cells from the neural folds and neural tube involves distinct mechanisms affecting both the tissue shape and the cell environment. Though these mechanisms are not fully understood at the molecular level, the analysis of the distribution of several molecules reported to promote intercellular contacts indicates a correlation between cell detachment from the neurepithelium and the decrease of cell adhesion molecule expression. During the following phase of migration, the mode of adhesion of crest cells switches preferentially to the cell-to-substratum type. Various experiments showed that among the major extracellular matrix components, fibronectins were decisive in promoting cell attachment, spreading and motility. Additional studies on receptors for fibronectins gave new insights on the differences between a motile and a stationary cell. These results will be discussed with particular reference to the migration at the cephalic level where most perturbation experiments were performed.

The molecular biology of fibronectins provided a finer understanding of the interactions between a cell and these molecules. The tools derived from this technique will open new areas of investigation hopefully leading to a better understanding of (i) the regulation of the cell-fibronectins interaction and (ii) the specificity of the pathways of migration followed by migrating cells, like the descendants of the neural crest.
β3 tubulin expression characterizes the differentiating mesodermal germ layer during Drosophila embryogenesis
Authors: Leiss Dagmar , Hinz Uwe , Gasch Alexander , Mertz Ronald , Renkawitz-Pohl Renate
Published: April 2021

During embryogenesis, the β3 tubulin gene of Drosophila is transcribed predominantly in the mesoderm. We have raised antibodies specific to the C-terminal domain of the β3 tubulin and analysed by immunostaining the distribution of this tubulin isotype during Drosophila embryogenesis. The protein is first detectable in the cephalic mesoderm at maximal germband extension. Shortly afterwards, β3 tubulin is expressed in single cells at identical positions of the thoracic and abdominal segments. We suggest that these cells represent muscle pioneer cells of Drosophila. During later embryonic development the somatic musculature, visceral musculature, dorsal vessel and macrophages contain β3 tubulin. In dorsalizing mutants dorsal, snail and twist, which do not form a ventral furrow during gastrulation, β3 expression is greatly reduced but not completely abolished. Our analysis shows that β3 tubulin immunostaining characterizes the differentiation of mesodermal derivatives during embryogenesis.
Development of functional innervation in the second and third order auditory nuclei of the chick
Authors: Pettigrew A. G. , Ansselin A. D. , Bramley J. R.
Published: April 2021

The neurones that constitute the auditory nuclei of the brainstem in the chick (nuclei magnocellularis, NM, and laminaris, NL) are generated between days 2 and 4 of incubation. These neurones migrate towards the dorsal surface of the brainstem over the next few days and reach their final destination at about day 9 of incubation. We have examined the development of functional connections between the auditory nerve and neurones in NM and between neurones in NM and NL in embryos from stage 34 (day 8 of incubation) using electrophysiological techniques and electron and light microscopy. The earliest extracellular recordings of electrically evoked field and spike potentials were made in NM with stimulation of the ipsilateral auditory nerve and in NL with stimulation of the ipsilateral NM at stage 35 (day 9). No activity could be recorded in NL with stimulation of the auditory nerve at this stage. By stage 37 (day 11), direct stimulation of the contralateral NM evoked responses in NL and by stage 38 (day 12) stimulation of the auditory nerves evoked stable field potentials in NL. These potentials changed polarity as the electrode penetrated NL in a direction that was perpendicular to the laminar arrangement of neurone somas and parallel to the dendritic axes of these neurones. In 18 of 26 analyses of current-source density in NL of 12 preparations between stages 38 and 40 there was a sink of current associated with synaptic activity at levels both above and below the source of current (cell somas) following stimulation of the ipsilateral auditory nerve. In the remaining analyses, and in all 15 analyses from preparations older than stage 40, stimulation of the ipsilateral input evoked only a single sink of current above the level of the cell somas. In all preparations from embryos at stage 38 and older, stimulation of the contralateral auditory nerve was associated with a single sink of current below the level of the cell somas. The axon projections to the ipsilateral NL from neurones in NM were examined using HRP labelling between stages 38 and 40. The presence of terminal fields of single axons in both the dorsal and ventral dendritic regions of the ipsilateral NL at these ages was confirmed. Furthermore, dense vesicles within synaptic terminals in both the dorsal and ventral dendritic fields could be identified in preparations at stage 36 following injection of HRP into NL and stimulation of the ipsilateral NM. The observations from these experiments suggest that the ventral dendrites in NL receive a transient functional innervation from neurones in the ipsilateral NM at early stages of development before the normal segregated innervation of NL is established.
Pioneer neurones use basal lamina as a substratum for outgrowth in the embryonic grasshopper limb
Authors: Anderson Hilary , Tucker Richard P.
Published: April 2021

During axonogenesis, contacts made by the growth cone with its substratum are important in guiding the direction of neurone outgrowth. This study examines the contacts made by the growth cones of pioneer neurones in the embryonic grasshopper limb. Individual pioneer neurones at different stages of development were injected with horseradish peroxidase and the contacts made by the filopodia at the tip of their growth cones were examined by electron microscopy.

Filopodia made few contacts with mesodermal cells, some contacts with ectodermal cells and very frequent contacts with basal lamina underlying the ectoderm. Components of the basal lamina may therefore play a role in guiding pioneer axon outgrowth.
Experimental manipulation of sexual differentiation in wallaby pouch young treated with exogenous steroids
Authors: Shaw G. , Renfree M. B. , Short R. V. , O W.-S.
Published: April 2021

We have investigated the effects of androgen or oestrogen treatment of female or male tammar wallabies from the day of birth, when the gonads are histologically undifferentiated, to day 25 of pouch life, when the gonads and the Wolffian and Müllerian ducts have differentiated and the testes have migrated through the inguinal canal.

Female tammars treated with testosterone propionate (24-50 mg kg−1 day−1) orally for 25 days had enlarged Wolffian and Müllerian ducts. Mammary and pouch development, however, was indistinguishable from that of control females. The treatment had no apparent effect on ovarian development, or on ovarian position in the abdomen. The phallus of males and females was similar in size, and neither experimental treatment had a significant effect on its size at day 25.

Male tammars treated with oestradiol benzoate (l-2-2-5mgkg−1day−1) orally for 25 days had gross hypertrophy of the urogenital sinus. Testicular morphology was abnormal; many of the germ cells appeared necrotic, the seminiferous tubules were of reduced diameter, and there were few Leydig cells and increased amounts of fibrous tissue between the tubules. The cortex of these gonads contained some areas which had an ovarian appearance, lacking tubules and containing numerous germ cells. The

Müllerian ducts of control males had regressed, but this was prevented by oestrogen treatment, suggesting an inhibition of either Müllerian Inhibiting Substance (MIS) production or its action. Normal testicular migration was inhibited in treated males; the testes remained high in the abdomen, similar in position to the ovaries of control females, whilst control males all had testes in the inguinal region. The gubernaculum and processus vaginalis of control males extended into the scrotum, but in treated males they terminated outside it. Oestrogen treatment had no effect on the size of the scrotum and did not induce mammary or pouch development.

These experiments show that marsupials, like eutherians, have a dual hormonal control of Wolffian and Müllerian development. By contrast, the initial development of the mammary glands, pouch, gubernaculum and scrotum does not appear to be under hormonal control and is therefore likely to be autonomous and dependent on genotype.
Development of segmentation in zebrafish
Authors: Kimmel Charles B. , Sepich Diane S. , Trevarrow Bill
Published: April 2021

Recent findings on the nature and origin of segmentation in zebrafish, Brachydanio rerio, are reviewed. Segmented peripheral tissues include the trunk and tail myotomes, that are derived from somitic mesoderm, and the pharyngeal arches that are derived from head mesoderm in addition to other sources. Two major regions of the central nervous system, the spinal cord and hindbrain, are also segmentally organized, as deduced from the distribution of identified neurones in both regions and by formation of neuromeres in the hindbrain that contain single sets of these neurones. Neural and mesodermal segments in the same body region can be related to one another by their patterns of motor innervation. This relationship is simple for the spinal/myotomal segments and complex for the hindbrain/pharyngeal arch segments. Development of the segments is also complex. Mesodermal and ectodermal progenitors have separate embryonic origins and indeterminate cell lineages, and the embryonic cells migrate extensively before reaching their definitive segmental positions. Results of heat-shock experiments suggest that development of myotomal and spinal segments are regulated coordinately in postgastrula embryos. Segmental patterning may be a relatively late feature of zebrafish embryonic development.
Regulation and putative function of the Drosophila gap gene Krüppel
Authors: Jäckle Herbert , Gaul Ulrike , Redemann Norbert
Published: April 2021

The Drosophila segmentation gene Krüppel (Kr) is expressed in a broad band of cells that covers about four-segment primordia in the blastoderm embryo. Examination of size and position of the Kr protein domain in various mutant embryos revealed that the establishment of the domain of Kr gene expression is under the control of the maternal effect pattern organizers which act at the poles. The lack of Kr activity causes a gap in the segment pattern of the embryo which is about twice the size of the Kr expression domain and extends posterior to it. This indicates that Kr activity per se is not directly responsible for the establishment of the pattern elements which are deleted in the mutant embryo. Examination of the molecular lesions in four Kr alleles indicated that each of them is a point mutant within the coding sequence of the Kr gene and each mutation results in a different replacement of a single amino acid within the ‘finger domain’ of the Kr protein. Thus, this region of the Kr protein is essential for Kr function. Since this portion of the Kr protein shares structural homology with the DNA-binding domain of several transcription factors, we propose that Kr acts as a transcription factor on subordinate genes that process the spatial cues provided by Kr activity to establish eventually the segments in the central region of the embryo.
The present status of the parasegment
Authors: Lawrence Peter A.
Published: April 2021

The hypothesis that parasegments are a fundamental unit of design in the Drosophila embryo was put forward three years ago (Martinez-Arias & Lawrence, Nature 313, 639-642, 1985). Here, I summarize new evidence in favour of the hypothesis.
Expression of Schwann cell markers by mammalian neural crest cells in vitro
Authors: Smith-Thomas L. C. , Fawcett J. W.
Published: April 2021

During embryonic development, neural crest cells differentiate into a wide variety of cell types including Schwann cells of the peripheral nervous system. In order to establish when neural crest cells first start to express a Schwann cell phenotype immunocytochemical tech niques were used to examine rat premigratory neural crest cell cultures for the presence of Schwann cell markers. Cultures were fixed for immunocytochemistry after culture periods ranging from 1 to 24 days. Neural crest cells were identified by their morphology and any neural tube cells remaining in the cultures were ident ified by their epithelial morphology and immunocyto chemically. As early as 1 to 2 days in culture, approxi mately one third of the neural crest cells stained with m217c, a monoclonal antibody that appears to recognize the same antigen as rat neural antigen-I (RAN-1). A similar proportion of cells were immunoreactive in cultures stained with 192-IgG, a monoclonal antibody that recognizes the rat nerve growth factor receptor. The number of immunoreactive cells increased with time in culture. After 16 days in culture, nests of cells, many of which had a bipolar morphology, were present in the area previously occupied by neural crest cells. The cells in the nests were often associated with neurones and were immunoreactive for m217c, 192-IgG and antibody to S-100 protein and laminin, indicating that the cells were Schwann cells. At all culture periods examined, neural crest cells did not express glial fibrillary acidic protein.

These results demonstrate that cultured premigratory neural crest cells express early Schwann cell markers and that some of these cells differentiate into Schwann cells. These observations suggest that some neural crest cells in vivo may be committed to forming Schwann cells and will do so provided that they then proceed to encounter the correct environmental cues during embry onic development.
Differential timing of nuclear lamin A/C expression in the various organs of the mouse embryo and the young animal: a developmental study
Authors: Röber Ruth-Ariane , Weber Klaus , Osborn Mary
Published: April 2021

In mouse embryos, acquisition of the nuclear lamin polypeptides A/C varies according to developmental stage and tissue type. In order to determine the precise time points and cell types in which lamin A/C are first observed, we have used two monoclonal antibodies in immunofluorescence studies of different tissues of developing mouse embryos and of young mice. One antibody (mAB346) is specific for lamins A and C, while the other (PKB8) detects lamins A, B and C. Dividing uterine development into three phases - germ layer formation, organogenesis and tissue differentiation - our results show that lamin A/C expression in the embryo proper is not observed until the third phase of development. Lamin A/C first appears at embryonic day 12 in muscle cells of the trunk, head and the appendages. Three days iater it is also seen in cells of the epidermis where its appearance coincides with the time of stratification. In the simple epithelia of lung, liver, kidney and intestine, as well as in heart and brain, lamins A/C do not appear until well after birth. Embryonal carcinoma (EC) cells express lamin B but not lamin A/C. Lamin A/C expression is noted in some EC cells after they are induced to differentiate and in several differentiated teratocarcinoma cell lines. Our results suggest that commitment of a cell to a particular pathway of differentiation (assayed by cell-type-specific expression of intermediate filament proteins) usually occurs prior to the time that lamin A/C can be detected. Thus lamin A/C expression may serve as a limit on the plasticity of cells for further developmental events.
PDGF receptors on cells of the oligodendrocyte-type-2 astrocyte (O-2A) cell lineage
Authors: Hart Ian K. , Richardson William D. , Heldin Carl-Henrik , Westermark Bengt , Raff Martin C.
Published: April 2021

It has been shown previously that cultures of rat optic nerve contain three types of macroglial cells - oligodendrocytes and two types of astrocytes. Type-1 astrocytes develop from their own precursor cells beginning before birth, while oligodendrocytes and type-2 astrocytes develop postnatally from a common bipotential precursor called the O-2A progenitor cell. Proliferating O-2A progenitor cells give rise to postmitotic oligodendrocytes beginning around birth, and to type-2 astrocytes beginning in the second postnatal week. Studies in vitro have suggested that platelet-derived growth factor (PDGF), secreted by type-1 astrocytes, plays an important part in timing oligodendrocyte development: PDGF seems to keep O-2A progenitor cells proliferating until an intrinsic clock in the progenitor cells initiates the process leading to oligodendrocyte differentiation. The clock apparently determines when a progenitor cell becomes unresponsive to PDGF, at which point the cell stops dividing and, as a consequence, automatically differentiates into an oligodendrocyte.

Here we have used radiolabelled PDGF to show that O-2A progenitor cells have PDGF receptors, suggesting that these cells respond directly to PDGF. The receptors resemble the type A PDGF receptor previously described on human fibroblasts and are initially retained when progenitor cells stop dividing and develop in vitro into oligodendrocytes. The latter finding indicates that receptor loss is not the reason that progenitor cells initially become mitotically unresponsive to PDGF.
Octopod, a homeotic mutation of the moth Manduca sexta, influences the fate of identifiable pattern elements within the CNS
Authors: Booker Ronald , Truman James W.
Published: April 2021

Octopod (Octo) is a mutation of the moth Manduca sexta, which results in the homeotic transformation of the ventral surface of the first (Al) and less often the second (A2) abdominal segments in the anterior direction. The extent of the transformation ranges from a slight deformation of the ventral cuticle, up to the formation of miniature thoracic legs on Al. The extent of the transformation is always less within A2 as compared to Al. A genetic analysis revealed that Octo is an autosomal mutation which shows incomplete dominance. The effect of this mutation on the central nervous system (CNS) was assessed by examining the distribution and fate of the postembryonic neuroblasts in the segmental ganglia of Octo larvae. In each of the thoracic ganglia of wildtype larvae, there is a set of 45–47 neuroblasts; a reduced but homologous array of 24 and 10 neuroblasts are found in Al and A2, respectively. Ganglion Al of Octo larvae had 1 to 6 supernumerary neuroblasts, and 20% of the A2 ganglia showed a single ectopic neuroblast. The supernumerary neuroblasts corresponded to identifiable neuroblasts normally found in more anterior ganglia. The Octo mutation also influenced the mitotic activity of stem cells normally present in Al. In this case, the neuroblasts generated a lineage of cells that were typical of a thoracic location rather than Al. These data demonstrate that homeotic mutations can influence the fate of identifiable pattern elements within the CNS of an insect.
Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice
Authors: Kothary Rashmi , Clapoff Susan , Darling Susan , Perry Marc D. , Moran Laurence A. , Rossant Janet
Published: April 2021

Transgenic mice have been generated that express the E. coli β-galactosidase gene under the control of the promoter from the mouse heat-shock gene, hsp68. Sequences from -664 to +113 relative to the start of transcription of the hsp68 gene were sufficient to direct stress-induced expression of the β-galactosidase gene in adult tail tissue and various tissues of fetal stages of development. Expression was detected in situ by staining with the chromogenic substrate, X-gal. The hybrid gene was refractory to induction in preimplantation embryos until the blastocyst stage of development, as reported for the endogenous hsp68 gene. No constitutive expression was observed by in situ staining or Northern analysis at any stage of development, even in tissues that constitutively express the endogenous hsp68 gene. We conclude that the hsp68 promoter region included in the construct contains sufficient sequence information for heat and arsenite inducibility, but it does not contain sequences controlling tissue-specific expression during development. This tightly regulated inducible promoter may provide a useful tool for short-term inducible gene expression in transgenic mice.
Biochemical research on oogenesis. RNA accumulation in the oocytes of the newt Pleurodeles waltl
Authors: Eynde Hilde Van Den , Mazabraud André , Denis Herman
Published: April 2021

We have compared the accumulation of 5S RNA and tRNA in oocytes of Pleurodeles waltl with the corresponding process previously studied in Xenopus laevis. 5S RNA synthesis is regulated similarly in both species since different families of 5S RNA genes are transcribed in oocytes and in somatic cells of P. waltl, as in those of X. laevis. Previtellogenic oocytes of P. waltl contain only one prominent kind of storage particles (thesaurisomes). In contrast, X. laevis oocytes of the same size contain two major classes of thesaurisomes, sedimenting at 42S and 7S. The more abundant particles found in P. waltl oocytes are homologous to the larger thesaurisomes (42S) of X. laevis, but they have a lower sedimentation coefficient and a higher tRNA/5S RNA molar ratio than their X. laevis counterparts. Small amounts of particles which we think to be homologous to the 7S particles of X. laevis are present in previtellogenic oocytes of P. waltl. Therefore, the storage function of the 7S particle protein (TFIIIA) is only marginal in this species. In X. laevis oocytes TFIIIA has a second function. It acts as a positive transcription factor involved in the developmentally regulated expression of the 5S RNA genes. In X. laevis expression of the oocyte-type 5S RNA genes is accompanied by a massive accumulation of TFIIIA. This is not the case in P. waltl.
A study of shaggy reveals spatial domains of expression of achaete–scute alleles on the thorax of Drosophila
Authors: Simpson Pat , Carteret Cathie
Published: April 2021

A study of shaggy mutant clones on the notum reveals that a greater number of cells are diverted into the bristle pathway of differentiation and fewer cells remain to produce the epidermis, shaggy clones differentiate supernumerary microchaetae and macrochaetae but these are found in the correct spatial locations, e.g. clusters of macrochaetae are formed round the position of the extant macrochaetae. The shaggy mutant phenotype requires the functioning of the genes of the achaete-scute (AS–C) complex but a dosage study shows that it is unlikely that the AS–C is overexpressed in shaggy cells. Data are presented that argue, also, for a correct spatial expression of the AS–C in shaggy mutants. A study of clones doubly mutant for shaggy and different achaete and scute alleles is consistent with the hypothesis that the clusters of macrochaetae formed by shaggy represent the restricted spatial domains of expression of the AS–C. The results can be reconciled with the known role for the AS–C, in determining which bristle types differentiate where, and a role for shaggy in the cell interactions, within domains of the AS–C expression, leading to the definition of only one bristle mother cell.
Paradoxical perpendicular contact guidance displayed by mouse cerebellar granule cell neurons in vitro
Authors: Nakatsuji Norio , Nagata Isao
Published: April 2021

In order to study migration of neurons in vitro, we cultured microexplants of the newborn mouse cerebellum outer layer, which is rich in immature granule cells, on a substratum double-coated with poly-L-lysine and laminin. The granule cells first migrated away from the explant along radially oriented parallel bundles of their neurites, thus displaying typical contact guidance. Then, in almost all explants, they changed their orientation by 90° to extend cell processes and translocate perpendicular to the radial neurites. Orientation and migration of neurons perpendicular to the aligned parallel structure is a novel type of contact-guided cell behavior, and may have interesting implications in migration of neurons in the cerebellum and other parts of the nervous system.
Prepupal differentiation in Drosophila: distinct cell types elaborate a shared structure, the pupal cuticle, but accumulate transcripts in unique patterns
Authors: Fechtel Kim , Fristrom Dianne K. , Fristrom James W.
Published: April 2021

The components of the pupal cuticle are the main differentiation products synthesized by both the larval and adult epidermis during the prepupal period of Drosophila development. The pupal cuticle is formed in vitro by imaginal discs in response to a 6 h pulse of 20-hydroxyecdysone (20-HE). We previously described the isolation and initial characterization of four ecdysone-dependent genes (EDGs) whose expression in imaginal discs occurs only in response to a pulse of 20-HE. In this report, we demonstrate that the pattern of temporal and tissue-specific expression of these EDGs in vivo is like that expected for genes that encode pupal cuticle proteins. Transcripts of these genes are detected in prepupae only in the epidermis and only when cuticle components are synthesized and secreted. Nonetheless, their temporal and spatial patterns of accumulation differ. EDG-84A-1 transcripts accumulate only in prepupae and only in imaginal cells. EDG-78E and EDG-64CD transcripts accumulate at the same time in both larval and imaginal cells. EDG42-A transcripts appear first in prepupae in imaginal cells and then, after a 2-4 h lag, in larval cells. It is evident that some genes are not restricted in their expression to only larval or imaginal epidermis.
Biochemical differentiation in a mutant of Dictyostelium discoideum defective in cyclic AMP chemotaxis and in intercellular cohesion
Authors: Srinivas Usha K. , Henderson Ellen J.
Published: April 2021

A temperature-sensitive mutant of Dictyostelium discoideum has been isolated based on its lack of chemotaxis toward cyclic AMP at the restrictive temperature, 27°C. The mutant develops normally at the permissive temperature, 22°C, but fails to aggregate or complete development at the restrictive temperature. The temperature-sensitive phenotype can be bypassed by allowing cultures to grow into late log phase or to starve for 60 –90 min at 22°C prior to a shift to 27°C. At 27°C, the mutant overproduces cell surface cyclic AMP receptors of both high and low affinity and is capable of spontaneous oscillations in light scattering in cell suspensions.

Despite its complete lack of morphological development, the mutant undergoes extensive biochemical differentiation. At the onset of starvation, it shows increased levels of N-acetylglucosaminidase, it expresses cyclic AMP receptors at the normal time and, although somewhat slowly, suppresses those receptors as if aggregation had been achieved. Metabolic pulse labellings with [35S]methionine revealed that the mutant at 27 °C displays the same changes in the patterns of newly synthesized proteins observed during the vegetative-to-aggregation and the aggregation-to-slug stages of normal development. The only clear difference from wild type was the failure of the culmination-stage isozyme of β-glucosidase to appear.

The mutant is defective in establishment of intercellular cohesion mechanisms, correlated with poor agglutination by concanavalin A, at the restrictive temperature. The properties of the mutant place severe constraints on models regarding the role of chemoreception and intercellular cohesion in regulation of gene expression.
The development of the Xenopus retinofugal pathway: optic fibers join a pre-existing tract
Authors: Easter S. S. , Taylor J. S. H.
Published: April 2021

The developing optic nerve and tract have received considerable attention in recent years, but the cellular and subcellular microenvironment of the growing axons has not been described. In the belief that such a description is essential (though certainly not sufficient) for an understanding of pathway formation, we have examined the normal development of the retinofugal projection of Xenopus laevis.

Optic fibers were labeled anterogradely at the retina with horseradish peroxidase (HRP) or the carbocyanine dye, Dil, at stages 32 to postmetamorphosis. The brains were examined both as whole mounts and in sections, light- and electron-microscopically, with the emphasis on tracts associated with the route of the optic fibers.

At stage 32, two ventral commissures were present, the anterior and postoptic. They were immediately subjacent to the pia. All tracts and even isolated axons were in similarly superficial locations. The first deep pathway (separated from the pia by cell nuclei) was seen at stage 46; it was a dorsal commissure, probably the posterior.

The first retinal axons passed from the optic stalk into the ventral part of the diencephalon, where they coursed along the rostral edge of the postoptic commissure, and maintained this position, relative to the other fibers in the tract of the commissure, throughout the remainder of their contralateral trajectory. They reached the presumptive thalamic and tectal termination sites and arborized. Subsequent optic axons followed this same route, thus enlarging the optic pathway relative to the more slowly growing nonoptic part of the commissure and its tract. Electron microscopy revealed, as early as stage 35, specialized contacts between cellular processes in the neuropil. These contacts had the form of symmetric membranous thickenings; some were associated with vesicles and were presumed to be synapses.

We conclude that the early forebrain and midbrain have only two ventral commissural pathways, and most axons that grow out after these pathways have formed add to them rather than establish new tracts. The optic axons travel a stereotyped pathway alongside a preexisting tract associated with the postoptic commissure. The possibility that optic fiber outgrowth is normally influenced by pre-existing tracts is discussed in relation to recent experimental investigations of fiber growth from ectopic eyes.
Complex spatio-temporal accumulation of alternative transcripts from the neurogenic geneDeltaduringDrosophilaembryogenesis
Authors: Kopczynski Casey C. , Muskavttch Marc A. T.
Published: April 2021

Delta (Dl) function is required for proper specification of epidermal and neural lineages within the neurogenic ectoderm of Drosophila melanogaster. We have determined the spatial accumulation of five Dl transcripts that arise as the result of alternative RNA processing during embryogenesis. We find that these transcripts accumulate in all cells of the neurogenic ectoderm immediately preceding neuroblast segregation, indicating that transcription of Dl does not differ between presumptive neuroblasts and presumptive dermoblasts. Dl transcripts also accumulate transiently in mesodermal and endodermal cells, suggesting that Dl may function in developmental processes in addition to differentiation of the neurogenic ectoderm. We find that three of the Dl transcripts are localized to the base of the nucleus during cellularization. The apparent association of these three transcripts with polysomes suggests that they accumulate within the cytoplasm at the nuclear periphery and is consistent with the hypothesis that Dl encodes multiple translational products.
Effect of the notochord on proliferation and differentiation in the neural tube of the chick embryo
Authors: van Straaten H. W. M. , Hekking J. W. M. , Beursgens J. P. W. M. , Terwindt-Rouwenhorst E. , Drukker J.
Published: April 2021

After implantation of a notochord fragment lateral to the neural tube in a 2-day chick embryo, at 4 days the ipsilateral neural tube half was increased in size and axons left the neural tube in a broad dorsoventral area (van Straaten et al. 1985). This enlargement appears to coincide with an increased area of AChE-positive basal plate neuroblasts, as determined with scan-cytophotometry. The effect was ipsilateral and local: clear effects were seen only when the implant was localized less than 80 μm from the neural tube and over 120 μm from the ventral notochord. In order to investigate the expected enhancement of proliferation, the mitotic density and the number of cells at the site of the implant at 3 days was determined and the mitotic index calculated. All three parameters showed an increase. It was coneluded that the cell cycle was shorter in the implant area relative to the control area, at least during the third day. At 4 days the number of cells was still increased, predominantly in the basal plate. It appeared that the numerical increase was for the larger part due to neuroblasts. The synergism of two notochords thus resulted in enhancement of proliferation and differentiation in the neural tube. It is suggested that the notochord merely regulates and arranges the surrounding sclerenchymal cells, which are the effective cells in the regulation of neural tube development.