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PubMed2006

Pristionchus pacificus: a well-rounded nematode.

Nematodes pervade Earth's biosphere and occupy innumerable ecological niches. The role of Caenorhabditis elegans as a model for developmental processes has encouraged us to cultivate a second nematode, Pristionchus pacificus, as a comparative counterpoint to address questions in development, behavior and ecology in nematode evolution. We hope that this endeavor, now more than a decade underway, will allow us to project findings onto other comparative models for biological processes. To this end, our laboratory has made an extensive genetic map and mutant screens to understand changes in developmental programs. Recently, we have been capitalizing on the whole genome sequence of P. pacificus to describe more thoroughly the molecular basis for these changes, as well as to better integrate our molecular knowledge with the biodiversity of Pristionchus species.

Animals↗

PubMed2002

Transient production of alpha-smooth muscle actin by skeletal myoblasts during differentiation in culture and following intramuscular implantation.

alpha-smooth muscle actin (SMA) is typically not present in post-embryonic skeletal muscle myoblasts or skeletal muscle fibers. However, both primary myoblasts isolated from neonatal mouse muscle tissue, and C2C12, an established myoblast cell line, produced SMA in culture within hours of exposure to differentiation medium. The SMA appeared during the cells' initial elongation, persisted through differentiation and fusion into myotubes, remained abundant in early myotubes, and was occasionally observed in a striated pattern. SMA continued to be present during the initial appearance of sarcomeric actin, but disappeared shortly thereafter leaving only sarcomeric actin in contractile myotubes derived from primary myoblasts. Within one day after implantation of primary myoblasts into mouse skeletal muscle, SMA was observed in the myoblasts; but by 9 days post-implantation, no SMA was detectable in myoblasts or muscle fibers. Thus, both neonatal primary myoblasts and an established myoblast cell line appear to similarly reprise an embryonic developmental program during differentiation in culture as well as differentiation within adult mouse muscles.

Actins↗

PubMed2004

Axonogenesis in the medaka embryonic brain.

In order to know the general pattern of axonogenesis in vertebrates, we examined axonogenesis in the embryonic brain of a teleost fish, medaka (Oryzias latipes), and the results were compared with previous studies in zebrafish and mouse. The axons and somata were stained immunocytochemically using antibodies to a cell surface marker (HNK-1) and acetylated tubulin and visualized by retrograde and anterograde labeling with a lipophilic dye. The fiber systems developed correlating with the organization of the longitudinal and transverse subdivisions of the embryonic brain. The first axons extended from the synencephalic tegmentum, forming the first fiber tract (fasciculus longitudinalis medialis) in the ventral longitudinal zone of the neural rod, 38 hours after fertilization. In the neural tube, throughout the entire brain two pairs of longitudinal fiber systems, one ventral series and one dorsal or intermediate series, and four pairs of transverse fiber tracts in the rostral brain were formed sequentially during the first 16 hours of axon production. In one of the dorsal longitudinal tracts, its branch retracted and disappeared at later stages. One of the transverse tracts was found to course in the telencephalon and hypothalamus. The overall pattern of the longitudinal fiber systems in medaka brain is similar to that in mouse, but apparently different from that in zebrafish. We propose that a ventral tract reported in zebrafish partially belongs to the dorsal fiber system, and that the longitudinal fiber systems in all vertebrate brains pass through a common layout defined by conserved genetic and developmental programs.

Animals↗

PubMed1987

Transection of the infraorbital nerve in newborn hamsters alters the somatosensory but not the visual representation in the superior colliculus.

The experiments described in this report were designed to determine whether changing the somatosensory representation in the deep laminae of the hamster's superior colliculus would result in a corresponding reorganization in the visual map in the overlying superficial layers. The somatosensory representation was altered by transecting the infraorbital (IO) nerve on the day of birth. This trigeminal branch supplies, among other targets, the snout and mystacial vibrissa follicles. These peripheral structures compose a major portion of the somatosensory representation in the deep collicular laminae. The effects of these lesions were assessed in anatomical and physiological experiments when the animals reached adulthood. Retrograde tracing with true blue demonstrated a 48% reduction in the number of trigeminocollicular neurons in the partially deafferented subnucleus interpolaris (the main source of trigeminal input to the rodent colliculus--e.g., Killackey and Erzurumlu: J. Comp. Neurol. 201:221-242, '81), but anterograde tracing with horseradish peroxidase (HRP) and wheat germ agglutinin-conjugated HRP showed that the terminal field of the trigeminocollicular projection from the deafferented subnucleus was essentially normal. This pathway terminated as a series of patches along the border between the stratum griseum intermediale and stratum album intermedium and encompassed approximately 75% of the rostrocaudal extent of the colliculus. The electrophysiological experiments revealed a marked change in somatosensory collicular topography. There was an under-representation of the vibrissae and the entire IO peripheral field in the deep laminae of the nerve-damaged animals, and neurons with trigeminal ophthalmic and mandibular receptive fields were recorded from portions of the colliculus in which only whisker-sensitive neurons would be normally isolated. There were no corresponding changes in the organization of the visual representation. These results support the conclusion that the organization of the visual representation in the superficial laminae and the organization of the somatosensory map in the deep layers of the mammalian superior colliculus follow independent developmental programs.

Animals↗

PubMed1991

Molecular milestones that signal axonal maturation and the commitment of human spinal cord precursor cells to the neuronal or glial phenotype in development.

Insights into the programmatic induction of neuronal and glial genes during human embryogenesis have depended largely on extrapolations of data derived from experimental mammals. However, the assumptions upon which these extrapolations are based have not been rigorously tested. Indeed, practically no information is available even on the human counterparts of the relatively small subset of well-characterized, developmentally regulated neuron and glial specific genes of the mammalian CNS. Thus, the developmental programs upon which human neural embryogenesis are based remain largely undeciphered. We have addressed this problem in immunohistochemical studies conducted on 22 human fetal spinal cords with gestational ages (GAs) that ranged from 6 to 40 weeks by using monoclonal antibodies to several classes of neuron or glial specific polypeptides. These polypeptides included: representatives of four different types (Types I-IV) of intermediate filament proteins, i.e., vimentin filament protein (VFP), glial fibrillary acidic protein (GFAP), different phospho-isoforms of the high (NF-H), middle (NF-M), and low (NF-L) molecular weight (Mr) neurofilament (NF) subunits, both acidic and basic cytokeratin (CK) proteins; three different microtubule associated proteins (MAPs), i.e., MAP2, MAP5, and tau; two different synaptic or coated vesicle proteins, i.e., synaptophysin (SYP) and clathrin light chain B (LCb); an oligodendroglial specific protein, i.e., myelin basic protein (MBP); and a receptor for a CNS trophic factor, i.e., the nerve growth factor receptor (NGFR).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1992

The rat's postero-orbital sinus hair: II. Normal morphology and the increase in peripheral innervation with adjacent nerve section.

The morphology and innervation of the postero-orbital (PO) sinus hair has been studied in normal rats and in adult animals in which an adjacent nerve, the infraorbital nerve, was sectioned on postnatal day 0 or day 7. The normal morphology of the follicle was similar to that of mystacial sinus hairs. However, the normal innervation differed from mystacial follicles in three respects: (1) instead of a separate innervation, the deep vibrissal nerve (DVN) and dermal plexus were supplied by a common follicle and skin nerve, named here the postero-orbital cutaneous nerve, a branch of the zygomaticofacial nerve; (2) the entry of the DVN through the capsule was highly variable; in some cases fascicles entered in close proximity, but in others they were widely distributed around the capsule; and (3) two or three small nerves, called here anastomosing nerves, were found to leave the PO follicle. These arose from the DVN after it had passed through the capsule to the cavernous sinus. The anastomosing nerves passed back through the capsule and ascended on the outer surface of the follicle to join the dermal plexus. Each nerve contained 1-4 myelinated fibres and 11-35 unmyelinated fibres. Infraorbital (IO) nerve section on day 0 caused a 19% (P less than 0.001, n = 8) increase in numbers of fibers to the DVN on the lesioned side. Most of the increase was due to unmyelinated fibres with no significant change in myelinated axons. No change in axon numbers in the DVN occurred after day 7 lesions. Labelling of the mystacial pad and the PO follicle did not result in any double labelling of cells in the trigeminal ganglion, in either normal or lesioned animals, making it improbable that the increased numbers of unmyelinated axons arose from rerouting of infraorbital fibres. It is suggested that the increased innervation of the PO follicle may arise by the rescue of ganglion cells from developmentally programmed cell death.

Animals↗

PubMed1995

Development of parvalbumin immunoreactivity in the chick Edinger Westphal nucleus.

To determine when the calcium-binding protein parvalbumin appears during development, neurons in the chick Edinger Westphal nucleus were examined for parvalbumin immunoreactivity at a variety of embryonic stages. Parvalbumin immunoreactivity appeared on embryonic day 14 (E14, Hamburger and Hamilton stage 40) in predominantly lateral Edinger Westphal neurons. Cytochrome oxidase activity within the nucleus was examined throughout development, as an indicator of physiological activity, and expression of cytochrome oxidase was compared with that of parvalbumin. Cytochrome oxidase activity was found to be uniformly high in all parts of the Edinger Westphal nucleus throughout development. Either the Edinger Westphal nucleus in physiologically active quite early in its development or other energy demands mask the correlation of cytochrome oxidase with electrical activity. Cytochrome oxidase was expressed well before parvalbumin immunoreactivity appeared. Voltage-activated calcium currents were characterized in E12 Edinger Westphal neurons. In both amplitude and composition, E12 calcium currents resemble those of E16 neurons, excluding the possibility that calcium currents appear de novo during or just prior to the appearance of parvalbumin. Both cytochrome oxidase activity and calcium currents are observed in Edinger Westphal neurons well before the appearance of parvalbumin during development. These findings do not exclude the possibility that physiological activity affects the expression of parvalbumin since other factors such as changing patterns of synaptic activity or the appearance of calcium conducting NMDA receptors have yet to be examined. However, they raise the possibility that additional factors such as an intrinsic developmental program or a change in the neuron's basal intracellular calcium requirements may also be involved.

Animals↗

PubMed1987

Different outcomes of synergy between song production and song perception in the same subspecies (Molothrus ater ater).

Here we report data requiring a reinterpretation of published results on the female cowbird's (Molothrus ater ater) ability to identify male song (King, A. P., and West, M. J. (1983). Female perception of cowbird song: A closed developmental program. Dev. Psychobiol., 16:335-342). Previously we stated that naive females, given extensive social experience with males from a distant cowbird subspecies, showed the same degree of preference for native song as did naive females reared with local males: extensive experience with another subspecies produced no evidence of postnatal modifiability. The procedure used to probe for modifiability did not, however, expose the different groups of females to comparable levels of stimulation because the females differentially affected the behavior of their male companions. We must therefore amend our conclusion about the modifiability of females. In addition, we report data on populational differences within the M.a. ater subspecies in the female's ability to influence heterosubspecific males, but data that are consistent with the more general pattern of synergistic effects during song development.

Animals↗

PubMed2006

Expression of the dlx gene family during formation of the cranial bones in the zebrafish (Danio rerio): differential involvement in the visceral skeleton and braincase.

We have used dlx genes to test the hypothesis of a separate developmental program for dermal and cartilage bones within the neuro- and splanchnocranium by comparing expression patterns of all eight dlx genes during cranial bone formation in zebrafish from 1 day postfertilization (dPF) to 15 dPF. dlx genes are expressed in the visceral skeleton but not during the formation of dermal or cartilage bones of the braincase. The spatiotemporal expression pattern of all the members of the dlx gene family, support the view that dlx genes impart cellular identity to the different arches, required to make arch-specific dermal bones. Expression patterns seemingly associated with cartilage (perichondral) bones of the arches, in contrast, are probably related to ongoing differentiation of the underlying cartilage rather than with differentiation of perichondral bones themselves. Whether dlx genes originally functioned in the visceral skeleton only, and whether their involvement in the formation of neurocranial bones (as in mammals) is secondary, awaits clarification.

Animals↗

PubMed1986

Developmental changes in the sensitivity of Volvox to ultraviolet light.

The response of Volvox to ultraviolet irradiation was analyzed. Young individuals isolated from a synchronous culture were exposed to UV light (120 J/m2) and subjected to variable length periods of dark following irradiation. The major effect of the UV treatment was the inability of the gonidia present in the colonies at the time of irradiation to continue and complete the developmental program. Individuals show a heightened sensitivity to UV for a limited period immediately following inversion and are insensitive at other stages of development. The cytotoxic effect of UV during this interval is completely reversed by the immediate exposure to white light and is increased with longer periods of dark treatment prior to exposure to white light. The temporal profile of the sensitivity defines a smooth curve in which the maximal sensitivity occurs three hours after inversion. The response to higher doses of UV (up to 500 J/m2) is a nonlinear increase in cytotoxicity and is disproportionately greater in those individuals just prior to the period of maximal sensitivity than those later in development. The results suggest that Volvox has at least two pathways for the repair of UV damage and that one of these, the principal dark repair pathway, is temporarily deficient in the gonidia of young individuals.

Animals↗

PubMed1988

The cyclic nucleotide phosphodiesterase of Dictyostelium discoideum: the structure of the gene and its regulation and role in development.

The cyclic nucleotide phosphodiesterase (phosphodiesterase) of Dictyostelium discoideum plays an essential role in development by hydrolyzing the cAMP used as a chemoattractant by aggregating cells. We have studied the biochemistry of the phosphodiesterase and a functionally related protein, the phosphodiesterase inhibitor protein, and have cloned the cognate genes. A 1.8-kb and a 2.2-kb mRNA are transcribed from the single-phosphodiesterase gene. The 2.2-kb mRNA comprises the majority of the phosphodiesterase mRNA found in differentiating cells and is transcribed only during development from a promoter at least 2.5 kb upstream of the translational start site. The 1.8-kb phosphodiesterase mRNA is detected at all stages of growth and development, is present at lower levels than the developmentally induced mRNA, and is transcribed from a site proximal to the protein-coding region. The phosphodiesterase gene contains a minimum of three exons, and a 2.3-kb intron, the longest yet reported for this organism. We have shown that the pdsA gene and four fgd genes affect the accumulation of the phosphodiesterase mRNAs, and we believe that these loci represent a significant portion of the genes regulating expression of the phosphodiesterase. The phosphodiesterase gene was introduced into cells by transformation and used as a tool to explore the effects of cAMP on the terminal stages of development. In cells expressing high levels of phosphodiesterase activity, final morphogenesis cannot be completed, and differentiated spore and stalk cells do not form. We interpret these results to support the hypothesis that cAMP plays an essential role in organizing cell movements in late development as well as in controlling the aggregation of cells in the initial phase of the developmental program.

3',5'-Cyclic-AMP Phosphodiesterases↗

PubMed1992

Mapping the mating type locus of Tetrahymena thermophila: meiotic linkage of mat to the ribosomal RNA gene.

Tetrahymena thermophila has a multiple mating type system. While a sexually mature cell usually expresses only one mating type, its germline (micronucleus) carries the genetic potential for 5 to 7 mating types. The set of allowed mating types is specified by the mat locus. The choice of which particular mating type is expressed by a cell reflects a somatically inherited, developmentally programmed differentiation of the somatic nucleus (macronucleus). In this work we report that the mat locus maps to the left arm of chromosome 2, as determined by nullisomic deletion mapping. We also report a distance of 29 cM between the mat locus and the ribosomal RNA gene, previously mapped to chromosome 2L. This represents another (rare) case of meiotic linkage in Tetrahymena.

Animals↗

PubMed1992

Development of sexual maturity in the ciliate Euplotes crassus: sources of variation in the timing of maturity.

The life styles of ciliated protists are particularly suitable for experimental analyses of certain aspects of developmental and genetic biology. The progression from sexual immaturity to maturity to senescence represents one of the most intriguing aspects of developmental programs. The extent to which progeny clones, their subclones, and testers used in the assay result in different lengths of immaturity has been investigated in Euplotes crassus. Six subclones from each of 12 progeny clones from a cross between stocks EC1 and EC2 were tested for maturity with stocks EC3, EC4, and EC5 on every transfer. Analysis of variance was used to partition the total variation in fissions to maturity into parts due to clones, subclones, and testers and the interactions between these levels. The error, interaction of subclones and testers, corresponds to a standard deviation of only 4.1 fissions, while the within clone within tester means range from 15.2 to 46.7 fissions; all levels except testers contribute significantly to the total variation. Most of the variability is attributable to clones (66%), the next most to error (16%), the next most to interaction of clones by testers (13%), and the least to subclones (5%). An a posteriori analysis examined whether the differences among clones were due to the cytoplasm of the clone ancestor (exconjugant), its mat (mating-type) locus genotype, or the mated pair it came from. None of these characteristics was able to interpret simply the large variability among clones. These results provide evidence that the transition from immaturity to maturity is quantitative and complex rather than a jump from one well-defined state to another.

Analysis of Variance↗

PubMed1989

Physical linkage of variable, diversity and joining gene segments in the immunoglobulin heavy chain locus of the mouse.

The variable region of antibody heavy chains is encoded by multiple variable (V), diversity (D) and joining (J) gene segments which seem to be assembled in a programmed developmental pathway during the maturation of B lymphocytes. A physical map linking V, D and J gene segments in the a haplotype of the murine Igh locus is presented in this report. The V gene located next to the known D and J gene segments maps 80 kb upstream of the JH locus and is a member of the VGAM3.8 V gene family. There is evidence for as yet unknown D gene segments located 5' to this V gene.

Animals↗

PubMed1990

Gamma/delta T cells and alpha/beta T cells differ in their developmental patterns of receptor expression and modulation requirements.

Measurement of the cell surface levels of gamma/delta (TcR 1) and alpha/beta (TcR 2) T cell receptors in the chicken revealed that thymocyte subpopulations that express these receptor isotypes differ remarkably in their rates of receptor acquisition. Whereas TcR 1 expression was relatively high (greater than 10(4) sites per cell) beginning on day 12 of embryonic life, the initial levels of TcR 2 expression on embryonic thymocytes were relatively low (approximately 10(3) sites per cell) when first measurable on day 15, and increased gradually as a function of T cell maturation. In peripheral tissues, the TcR 1 cells also expressed higher receptor levels than did the TcR 2 cells, but the difference was only 2-3-fold. The TcR 2 receptors on immature T cells could be easily modulated by receptor cross-linkage, very much like immunoglobulin receptors on immature B cells. While the TcR 2 receptor modulation occurred within minutes, TcR 1 receptor modulation required several hours for completion, even in the embryonic thymus. The data indicate very different developmental programs for TcR 1 and TcR 2 expression, and suggest fundamental differences in clonal selection modes for the two T cell subpopulations.

Animals↗

PubMed2004

Transient upregulation of Nkx2.2 expression in oligodendrocyte lineage cells during remyelination.

While numerous oligodendrocyte progenitor cells (OPCs) exist in the adult central nervous system (CNS), the molecular signals that promote or inhibit their differentiation into mature oligodendrocytes (OLs) are not known. To investigate whether remyelination in the adult CNS is regulated by the same mechanisms that promote developmental myelination, we used an acute demyelinating/remyelinating lesion in the adult rat spinal cord to examine the expression of the homeodomain transcription factor Nkx2.2, which has previously been implicated in oligodendrocyte differentiation during embryonic development. After a demyelinating insult, Nkx2.2 expression was upregulated first in NG2-expressing OPCs surrounding the lesion and subsequently in both precursors and OLs that appeared inside the lesion prior to the onset of remyelination. The temporal and spatial pattern of Nkx2.2 upregulation coincided with that of oligodendrocyte differentiation characterized in our previous study. A similar increase in the level of Nkx2.2 expression was observed in the postnatal developing optic nerve in a wave from the proximal to the distal retinal end. In vitro Nkx2.2 was expressed in OPCs and immature OLs isolated from postnatal rat spinal cord but was absent from mature OLs. These observations indicate that the process of generating new OLs in a remyelinating lesion recapitulates the developmental program involving activation of the Nkx2.2 gene, which may trigger the existing NG2-expressing precursors in the adult CNS to undergo terminal differentiation into remyelinating OLs.

Animals↗

PubMed1990

Region- and sex-related differences in maturation of astrocytes in dissociated cell cultures of embryonic rat brain.

Previous studies using dissociated cell cultures of fetal rat brain have revealed considerable regional diversity as well as sex steroid-independent sex differences in developmental schedules of dopaminergic neurons. Because these phenomena might be related to glial heterogeneity, cultures of dissociated male and female diencephalon, mesencephalon, and rhombencephalon of gestational day 14 rats were investigated with respect to the development of astrocytic markers. Cultures were incubated for 3-8 days in vitro (DIV) in serum-supplemented or serum-free medium. Vimentin and glial fibrillary acidic protein (GFAP) were quantified by counting of immunolabeled cells and immunoblotting. Vimentin and GFAP content rose from DIV 3 to 6 in all cultures. Regional variation of vimentin content was low, but large differences occurred in amounts of GFAP. GFAP reached high levels in rhombencephalon, especially when supplemented with serum, but remained very low or not detectable in mesencephalon. Simultaneous immunostaining for both cytoskeletal proteins revealed the presence of large numbers of vimentin single-labeled and small numbers of vimentin/GFAP double-labeled cells. Numbers of cells expressing GFAP showed similar regional variations as GFAP contents in both serum-free and serum-supplemented medium. They rose steeply from DIV 3 to 8 in rhomb- and diencephalon but not in mesencephalon. Transiently, female diencephalic cultures contained slightly more GFAP-immunoreactive cells than male cultures. The results thus demonstrate considerable regional heterogeneity of astrocytic maturation. However, neither the regional nor the sex differences show a consistent correlation with previous data on development of dopaminergic and other monoaminergic neurons in vitro. It seems likely that the dependence of neurons on glial environment for realization of an inherent developmental program varies among neuronal phenotypes.

Animals↗

PubMedDate not supplied

Cellular theories of aging as related to the liver.

The age-associated decline in function of several organs, including the liver, may be caused by mechanisms operating on the cellular level. Fibroblasts and several other cell types derived from normal individuals have limited lifespans in culture, and several abnormalities described for senescent cultured fibroblasts also apply to hepatocytes and other cell types obtained from aged organisms. Cellular theories of aging can be divided into two broad and overlapping categories: (a) those that view cell death as an actively programmed developmental process, and (b) those that consider cellular aging to result from a passive accumulation of errors in macromolecules. These theories are not necessarily mutually exclusive, and many of the phenotypic changes in senescent hepatocytes, fibroblasts and other cells are compatible with several different theories. The challenge for the future is to distinguish primary causes from secondary consequences of cellular aging so that rational attempts to intervene in the aging process are possible.

Aging↗