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Heat shock and thermotolerance in plant and animal embryogenesis.

Although the strategies of early embryogenesis differ greatly among multicellular eukaryotes, there are certain parallels in structure, form, and function that cross even kingdom lines: the extreme heat sensitivity of zygotes and very early embryos, followed by the acquisition of thermotolerance during subsequent development, is one such parallel. The heat sensitivity may be so extreme that even moderate increases in temperature result in lethality (generally associated with the earliest phases of embryogenesis), or the effects may be less severe, resulting in defects in development but not in lethality. Mechanistically, and molecularly, these two forms of thermosensitivity appear to have different origins. On the one hand, outright lethality appears to result from an inability to induce heat shock genes and proteins; on the other hand, heat-induced developmental defects appear to result from an alteration in expression of non-heat shock genes and from a delay in the overall developmental program that generally accompanies the cell's response to heat shock. This review is focused on the developmental regulation of the heat shock response during early embryogenesis and on the impact of this regulation on the development of both animal and plant embryos. The two basic issues that we address here are (i) the expression of heat shock genes in the absence of heat shock during embryogenesis and (ii) the expression (or lack of expression) of heat shock genes after deliberate exposure of the embryos to heat shock and the consequences of this expression on its subsequent survival and development.

Animals↗

Tissue-specific distribution and developmental regulation of M and B creatine kinase mRNAs.

To characterize the tissue-specific distribution and developmentally regulated expression of M and B creatine kinase mRNA in rats, total cellular RNA was isolated from adult rat tissues and from skeletal muscle, heart, brain and intestine at selected stages of development. Northern blots were prepared and hybridized with M and B subunit-specific probes derived from the 3'-untranslated region. M creatine kinase mRNA was expressed abundantly in heart and skeletal muscle, and less abundantly in lung. B creatine kinase mRNA was found in all tissues examined except liver and was abundant in brain, heart and intestine. The developmentally regulated expression of M and B creatine kinase mRNA was determined in skeletal muscle, heart, brain and intestine. The developmental program of B creatine kinase mRNA was different for each tissue examined. During development, M creatine kinase mRNA was up-regulated in both heart and skeletal muscle with a different regulatory program. This resulted in replacement of B mRNA by M mRNA as the predominant species at an earlier developmental stage in heart when compared to skeletal muscle.

Animals↗

Superinduction of the Dictyostelium discoideum cell surface cAMP receptor by pulses of cAMP.

Extracellular cAMP plays a crucial role in regulating the developmental program of Dictyostelium discoideum, functioning as a chemotactic agent, as well as a signal that regulates expression of developmentally expressed genes. These activities appear to be mediated by a cell-surface receptor for cAMP. We have studied the regulation of this receptor in cells developed in starved suspension cultures exposed to 50 nM pulses of cAMP every 6 min. cAMP-pulsed cells display roughly 10-fold higher cAMP receptor levels than cells that developed on filters or that were starved in suspension without cAMP pulses. Based on saturation binding analysis, the superinduced binding activity represents an increase in receptor number, while receptor affinity for cAMP is unaffected. Photoaffinity labeling of superinduced cells results in specific labeling of the same molecules that are labeled in starved cells. This increased cAMP binding activity was also detected in membrane preparations from cAMP-pulsed cells. These results provide evidence for an unusual mode of receptor regulation: autogenous induction of the receptor by its ligand.

Affinity Labels↗

Two transmembrane signaling mechanisms control expression of the cAMP receptor gene CAR1 during Dictyostelium development.

Dictyostelium discoideum is among the best characterized organisms for the study of receptor/guanine nucleotide binding protein-mediated control of differentiation. Dictyostelium grow unicellularly but form fully differentiated multicellular organisms through a developmental program regulated by secreted cAMP activating specific cell-surface receptors. Dictyostelium respond differentially to cAMP at different developmental stages. During early development, expression of certain genes is induced by low-level oscillations of extracellular cAMP. Later, continuous, high cAMP concentrations will promote expression of specific genes in multicellular structures. Here, we show that the cAMP receptor gene CAR1, which is essential for development, utilizes two promoters that are activated at distinct stages of development and respond to different extracellular cAMP conditions. One promoter is active with low-level oscillations of cAMP; exposure to high cAMP concentrations will repress this promoter and induce a second promoter. The CAR1 mRNAs are alternatively spliced but encode identical proteins. Thus, through differential sensitivity to its own ligand, cAMP, two promoters and alternative splicing regulate CAR1 expression during Dictyostelium development.

Alternative Splicing↗

Pre- and postnatal ontogeny of aromatase cytochrome P450 messenger ribonucleic acid expression in the male rat brain studied by in situ hybridization.

A central step in the development and sexual differentiation of the brain is the intraneuronal conversion of testosterone to estrogen. This conversion is catalyzed by an enzyme complex comprised of cytochrome P450 aromatase and the ubiquitous NADPH-dependent cytochrome P450 reductase. Brain aromatase activity is developmentally regulated and expressed only in specific hypothalamic and limbic structures. To determine whether developmental change in aromatase activity are the result of differential regulation of aromatase gene expression, we designed two specific oligonucleotides complementary to a 5'-region or to the heme-binding region of the rat aromatase complementary DNA. Relative amounts of messenger RNA (mRNA) encoding aromatase cytochrome P450 were examined on cryostat sections from male fetuses and offspring of time-pregnant Long-Evans rats using the in situ hybridization technique. On gestational day (GD) 16 (GD 1 = 24 h after mating), aromatase-specific mRNA was detected in the preoptic/hypothalamic area. On GD18 and GD20, aromatase mRNA expression was more abundant and more widespread. High levels of mRNA were detected in the medical preoptic nucleus, the sexually dimorphic nucleus of the preoptic area, the bed nucleus of stria terminalis, and the medial amygdala. Lower levels were found in the periventricular preoptic nucleus and the ventromedial hypothalamic nucleus. On postnatal day (PN) 2 and throughout further development (PN6, PN15, and adult), aromatase mRNA decreased to much lower levels in the preoptic area, but remained high in the bed nucleus of stria terminalis and amygdala. An intermediate situation, i.e. transient expression with a peak around birth, was observed for aromatase mRNA levels in the ventromedial hypothalamic nucleus. Interestingly, during early postnatal development, specific aromatase mRNA was also detected in the hippocampal formation as well as in the subfornical organ. In summary, these results demonstrate that the low quantities of aromatase transcripts in the developing rat brain can be detected by means of in situ hybridization. Depending on the particular region examined, aromatase expression appears to be subjected to different developmental programs. As the expression of aromatase mRNA parallels the results obtained from activity assays, it is likely that developmental changes in aromatase activity are the result of regulation of aromatase gene expression.

Animals↗

Development gene networks and evolution.

Animal development relies on complex programs of gene regulation that are likely to account for a significant fraction of the information carried in genomes. The evolution of these regulatory programs is a major contributor to the diversity of animal forms, yet the architecture of the transcriptional networks that comprise developmental programs is only beginning to come into focus. The sea urchin offers an uncomplicated system in which to study transcriptional regulation and the networks that direct embryogenesis. This review describes the approaches that we are taking towards this problem using the sea urchin embryo and some comparative methods that will eventually lead to a deeper understanding of the evolution of developmental networks.

Animals↗

The role of elav-like genes, a conserved family encoding RNA-binding proteins, in growth and development.

The elav gene is the prototype for a developmental regulator encoding a tissue-specific RNA-binding protein. The family of elav-like genes includes multiple members that are expressed in the nervous system at different times during development and also one that is expressed in all tissues. The elav-like proteins are proposed to function by binding to specific mRNAs and regulating their expression to control a developmental program. Consistent with this model, the overexpression of some elav-like genes results in an alteration in the developmental fates of some cells. Potential mRNA targets for these proteins have been identified along with functions in alternative RNA processing, cytoplasmic polyadenylation, and mRNA stability. This diversity of post-transcriptional regulatory events may be due to combinatorial interactions with other RNA-binding proteins and multiple effectors of RNA regulation.Copyright 1997 Academic Press Limited

Journal Article↗

Potential role of NKG2D/MHC class I-related chain A interaction in intrathymic maturation of single-positive CD8 T cells.

The nonclassical MHC class I molecule MHC class I-related chain A (MICA) interacts with the NKG2D receptor expressed at the surface of most peripheral CD8 T cells, gammadelta T cells, and NK cells. We investigated the role of MICA-NKG2D interactions in the selection or maturation of the T cell repertoire within the thymus using MICA tetramers and anti-MICA mAbs. MICA tetramers identified a small population of late stage CD8 single-positive, CD45RA(+) CD62L(+) CCR7(+) CD69(-) thymocytes, a phenotype compatible with that of fully mature CD8(+) cells ready to emigrate to the periphery as naive cells. MICA molecules were expressed in the outer layer of Hassal's corpuscles within the medulla of normal thymus. In thymomas, an overexpression of MICA in cortical and medullar epithelial cells was observed. This was associated with a decreased percentage of NKG2D-positive thymocytes, which expressed a less mature phenotype than in normal thymus. These results indicate that CD8(+) thymocytes up-regulate NKG2D as they complete their developmental program before leaving the thymic medulla to seed the periphery, and identify NKG2D as a potential regulator of the developmental processes in T cells that are essential for immune homeostasis.

Adult↗

Cell-surface area codes: mobile-element related gene switches generate precise and heritable cell-surface displays of address molecules that are used for constructing embryos.

We present an updated area code hypothesis supporting the proposal that cell surface display of seven-transmembrane olfactory receptors, protocadherins and other cell surface receptors provide codes that enable cells to find their correct partners as they sculpture embryos. The genetic mechanisms that program the expression of such displays have been largely unknown until very recently. However, increasing evidence now suggests that precise developmental control of the expression of these genes during embryogenesis is achieved in part by permanent and heritable changes in DNA. Using the developing immune system as a model, we discuss two different types of developmentally programmed genetic switches, each of which relies on recombination mechanisms related to mobile elements. We review new evidence suggesting the involvement of mobile element related switch mechanisms in the generation of protocadherin molecules, and their possible involvement in the control of expressions of olfactory receptors. As both recombinase and reverse transcriptase mechanisms play a role in the switching of the immunoglobulin genes, we searched the databases of expressed sequence tags (dbEST) for expression of related genes in other tissues. We present data revealing that transposases and reverse transcriptases are widely expressed in most tissues. We also searched these databases for expression of env (envelope) gene products, stimulated by provocative results suggesting that these molecules might function as cellular address receptors. We found that env genes are also expressed in large numbers in normal human tissues. One must assume that these three different types of mobile-element-related messenger RNA molecules (transposases, reverse transcriptases, and env proteins) are expressed for use in functions of value in the various tissues and have been preserved in the genome because of their selective advantages. We conclude that it is possible that many specific cell lineage decisions are made and remembered by means of genetic switches similar to those that control the immunoglobulin and protocadherin and, probably, the seven transmembrane/olfactory gene families. We also conclude that complex genetic programs utilizing mobile-element-related genes program these events.

Cell Membrane↗

Role of replication time in the control of tissue-specific gene expression.

Late-replicating chromatin in vertebrates is repressed. Housekeeping (constitutively active) genes always replicate early and are in the early-replicating R-bands. Tissue-specific genes are usually in the late-replicating G-bands and therein almost always replicate late. Within the G-bands, however, a tissue-specific gene does replicate early in those cell types that express that particular gene. While the condition of late replication may simply be coincident with gene repression, we review evidence suggesting that late replication may actively determine repression. As mammals utilize a developmental program to Lyonize (facultatively heterochromatinize) whole X chromosomes to a late-replicating and somatically heritable repressed state, similarly another program seems to Lyonize individual replicons. In frogs, all genes begin embryogenesis by replicating during a very short interval. As the developmental potency of embryonic cells becomes restricted, late-replicating DNA gradually appears. This addition to the repertoire of gene control--i.e., repression via Lyonization of individual replicons--seems to have evolved in vertebrates with G-bands being a manifestation of the mechanism.

Animals↗

"Alien" wasps and evolution of development.

A comparative analysis of early developmental programs in a group of parasitic wasps reveals that closely related species can undergo dramatic evolutionary shifts in their patterns of embryogenesis. Developmental changes detected include alterations in early cleavage divisions, the establishment of embryonic anteroposterior polarity and modifications of the segmentation gene hierarchy described from Drosophila. These changes appear to be adaptations to parasitic development, taking place within the body of the host. Wasps illustrate a surprising plasticity in their early development and embryogenesis. The alterations associated with different parasitic strategies suggest that ecological adaptations may have profound influences on developmental processes in animals.

Animals↗

Early restriction of the human antibody repertoire.

Diversification of the antibody repertoire in mammals results from a series of apparently random somatically propagated gene rearrangement and mutational events. Nevertheless, it is well known that the adult repertoire of antibody specificities is acquired in a developmentally programmed fashion. As previously shown, rearrangement of the gene segments encoding the heavy-chain variable regions (VH) of mouse antibodies is also developmentally ordered: the number of VH gene segments rearranged in B lymphocytes of fetal mice is small but increased progressively after birth. In this report, human fetal B-lineage cells were also shown to rearrange a highly restricted set of VH gene segments. In a sample of heavy-chain transcripts from a 130-day human fetus the most frequently expressed human VH element proved to be closely related to the VH element most frequently expressed in murine fetal B-lineage cells. These observations are important in understanding the development of immunocompetence.

Adult↗

Starvation promotes Dictyostelium development by relieving PufA inhibition of PKA translation through the YakA kinase pathway.

When nutrients are depleted, Dictyostelium cells undergo cell cycle arrest and initiate a developmental program that ensures survival. The YakA protein kinase governs this transition by regulating the cell cycle, repressing growth-phase genes and inducing developmental genes. YakA mutants have a shortened cell cycle and do not initiate development. A suppressor of yakA that reverses most of the developmental defects of yakA- cells, but none of their growth defects was identified. The inactivated gene, pufA, encodes a member of the Puf protein family of translational regulators. Upon starvation, pufA- cells develop precociously and overexpress developmentally important proteins, including the catalytic subunit of cAMP-dependent protein kinase, PKA-C. Gel mobility-shift assays using a 200-base segment of PKA-C's mRNA as a probe reveals a complex with wild-type cell extracts, but not with pufA- cell extracts, suggesting the presence of a potential PufA recognition element in the PKA-C mRNA. PKA-C protein levels are low at the times of development when this complex is detectable, whereas when the complex is undetectable PKA-C levels are high. There is also an inverse relationship between PufA and PKA-C protein levels at all times of development in every mutant tested. Furthermore, expression of the putative PufA recognition elements in wild-type cells causes precocious aggregation and PKA-C overexpression, phenocopying a pufA mutation. Finally, YakA function is required for the decline of PufA protein and mRNA levels in the first 4 hours of development. We propose that PufA is a translational regulator that directly controls PKA-C synthesis and that YakA regulates the initiation of development by inhibiting the expression of PufA. Our work also suggests that Puf protein translational regulation evolved prior to the radiation of metazoan species.

Amino Acid Sequence↗

Programmed cell death in prokaryotes.

Programmed cell death (PCD), also referred to as apoptosis, is a cellular "suicide" mechanism, based on information from its own internal metabolism, environment, developmental history, and genome. This system was described in eukaryotes continuously along evolution, through amoebae, nematodes, insects, and animals. PCD is essential for the proper development or function of a cell system, organ, or survival of the organism as a whole. Research in the last 2 decades has shown that the life cycle of several prokaryotic organisms display developmental programs, similar to metazoan differentiation, that is part of their adaptation to stressful environments. These include warmer cell formation and differentiation in Caulobacter cereus, sporulation in Bacillus and Streptomyces, heterocyst formation in Anabaena, development of bacteroids in Rhizobium, the formation of multicellular fruiting bodies and sporulation in Myxobacteria, and the formation of nonculturable, but viable, cells in various Gram-negative bacteria. Moreover, and more significantly, the photosynthetic bacteria Rhodobacter capsulatus were shown to release nucleoprotein particles designated "gene transfer agent (GTA)" as they enter the stationary phase. GTAs contain DNA of 3.6 x 10(6) molecular weight, representing all parts of the genome, and they may be taken up by other strains of R. capsulatus, and complement mutants. We postulate that these various modes of stress adaptations in bacteria are prokaryotic manifestation, and possibly the phylogenetic precursor, of the eukaryotic phenomenon, programmed cell death, and therefore we propose to designate it "proapoptosis". In addition to their function, apoptosis and proapoptosis share various mechanistic programmed features, including DNA fragmentation and packaging, cell shrinkage, degradation of RNA, proteolysis and synthesis of new proteins, and the involvement of reactive oxygen species.

Apoptosis↗

Retroviral transduction of TLS-ERG initiates a leukemogenic program in normal human hematopoietic cells.

Many chimeric oncogenes have been identified by virtue of the association between chromosomal translocation and specific human leukemias. However, the biological mechanism by which these oncogenes disrupt the developmental program of normal human hematopoietic cells during the initiation of the leukemogenic process is poorly understood due to the absence of an appropriate experimental system to study their function. Here, we report that retroviral transduction of TLS-ERG, a myeloid leukemia-associated fusion gene, to human cord blood cells results in altered myeloid and arrested erythroid differentiation and a dramatic increase in the proliferative and self-renewal capacity of transduced myeloid progenitors. Thus, TLS-ERG expression alone induced a leukemogenic program that exhibited similarities to the human disease associated with this translocation. These results provide an experimental examination of the early stages of the human leukemogenic process induced by a single oncogene and establish a paradigm to functionally assay putative leukemogenic genes in normal human hematopoietic cells.

Cell Lineage↗

Transient genetic asymmetry and cell fate in a bacterium.

Certain species of Gram-positive bacteria can initiate a developmental program that results in the formation of two daughter cells with different fates. One cell develops into a spore and the other cell undergoes programmed lysis, with each process being mediated by a cascade of cell-type-specific transcription factors. An early and critical step in this developmental pathway is the formation of a division septum near one pole, creating two compartments of different sizes. But how is this morphological asymmetry translated into the transcriptional asymmetry of the two compartments? Recent results suggest that the chromosomal position of the genes encoding several key components of the transcriptional regulatory network has an important role in this process.

Bacillus subtilis↗

Gametogenic processes and their relationship to normal and abnormal conceptus development.

The relationship between gametogenesis and conceptus development has been described in relation to recent experiments concerning selected aspects of the cellular and molecular basis of differentiation of female and male gametes. Evidence suggests that a variety of hormonal and nonhormonal mechanisms or combinations thereof have evolved and control distinct stages of oocyte maturation process. Analyses of the meiotic maturation process with particular emphasis on amphibians, indicate that distinct cytoplasmic factors are produced or activated in response to a hormonal stimulus which initiates germinal vesicle breakdown and reinitiates the meiotic maturation process. Evidence suggests that certain cytoplasmic factors originate in the nucleus and (or) cytoplasm and that considerable cytoplasmic maturation proceeds in the absence of the nucleus, or as a result of nuclear-cytoplasmic interactions. Cytoplasmic factor(s) or activities affect many aspects of oocyte function and structure, including the cell membrane, chromosomes, nucleus and meiotic spindle. Ions also play a crucial role in gamete differentiation either alone or in combination with hormones or these cytoplasmic factors. The relevance of these findings to oocyte maturation and fertilization, activation and embryonic development is discussed. In many cases, similar types of biological activities exist in widely separated species and (or) are effective in widely different species. The importance of synchronization of normal gametes to the development of the embryos is discussed. It is proposed, and the evidence suggests, that hormones, ions and cytoplasmic factors play a fundamental and variable role in the differentiation and function of "fully" mature oocytes of many species including mammals. Significantly, the data suggest that a "fully" mature oocyte occurs as a result of the expression of an underlying developmental program. Variations in these substances or the processes involved in their formation or actions appear to be relevant to understanding a wide variety of developmental abnormalities as well as to assessing the normality or abnormality of in vivo or in vitro maturation and differentiation.

Animals↗

A conserved partner-switching system controls terminal differentiation in multicellular cyanobacteria.

UNLABELLED: Canonical partner-switching systems (PSSs) regulate sigma factor activity through reversible phosphorylation, but their established roles have been largely limited to stress responses and sporulation in Firmicutes. Whether this regulatory mechanism also controls developmental cell fate decisions in other bacterial phyla has remained unknown. Here, we identify a canonical PSS that governs heterocyst differentiation in the multicellular cyanobacterium Anabaena sp. PCC 7120. This system comprises the anti-sigma factor All2284 (NfsS) and the anti-anti-sigma factor All2283 (NfsR). Structural predictions and biochemical assays showed that NfsS phosphorylates NfsR on a conserved serine residue, whereas bacterial two-hybrid and co-purification assays demonstrated that NfsS binds the developmental sigma factors SigC and SigE. Deletion of nfsR abolished heterocyst formation and diazotrophic growth, and transcriptomic analysis revealed broad failure to induce late heterocyst genes, including nitrogen fixation functions such as nifHDK and fdxH. Phylum-wide comparative genomics further showed that PSS genes and putative functional clusters are strongly enriched in filamentous and heterocyst-forming taxa, indicating an association between the expansion of these signaling modules and the emergence of multicellularity and developmental specialization. Together, these findings establish a PSS as a direct regulator of terminal cell differentiation in a gram-negative bacterium and reveal partner switching as a conserved regulatory principle linking environmental signaling to developmental fate in a major bacterial phylum. IMPORTANCE: While partner-switching systems are classically associated with stress responses and sporulation control in Firmicutes, whether this regulatory logic governs developmental decisions in other bacterial phyla has remained unknown. Here, we establish that a related partner-switching mechanism operates in a distinct bacterial lineage, the cyanobacteria, where it controls a major developmental transition involving terminal cell differentiation. Specifically, we show that a phosphorylation-dependent checkpoint involving the anti-sigma factor NfsS and the anti-anti-sigma factor NfsR directly regulates heterocyst formation. Disruption of this switch abolishes cell differentiation and diazotrophic growth, revealing that this system is an obligate gatekeeper for terminal differentiation. Conceptually, these findings substantially extend the known functional repertoire of partner-switching circuits: rather than controlling stress adaptation or spore dormancy, this module has been co-opted to govern a complex, multicellular developmental program in an organism that underpins global carbon and nitrogen cycles. This work, therefore, establishes a new paradigm for phosphorylation-based control of developmental sigma factors and provides a tractable model for dissecting how conserved signaling modules are rewired to drive lineage-specific innovations across the bacterial domain.

cell differentiation↗