Development of gabaergic connections in vitro: Increasing efficacy of synaptic transmission is not accompanied by changes in miniature currents

Development of inhibitory synaptic transmission was studied using a dissociated cell culture from the superior colliculus of neonatal rat. Patch-clamp recordings in the whole-cell configuration were performed to measure evoked (single-cell-activated) inhibitory postsynaptic currents (IPSCs), miniature IPSCs and current responses to maximal concentrations of exogenous γ-aminobutyric acid (GABA). Over a period of 3 weeks in vitro (DIV3-24), the fraction of synaptically coupled neurons raised from 0% to 76%. Evoked IPSCs were first observed at DIV5. They had an average amplitude of 33.9 pA during the first week (n = 13) and 129.7 pA during the fourth week (n = 48). This increase by a factor of 3.8 represents a significant rise in the efficacy of GABAergic transmission during in vitro development. However, no developmental change has been observed in the average amplitudes of miniature somatic IPSCs. The latter remained at an average level of about 9 pA (symmetrical chloride concentration and a driving force of 68 mV). No increase was found also in whole-cell current densities induced by saturating concentrations of exogenous GABA. Our results suggest that under the given conditions, synapse maturation was primarily the result of presynaptic sprouting. This conclusion is further supported by bouton counts in immunostained collicular cultures, where the number of axosomatic and axodendritic GABAergic contacts per neuron increased from 0.54 and 0.37, respectively, at DIV3, to 13.84 and >23.1, at DIV24. The overall density of GABAergic neurons decreased during this period from about 41,000/cm² to 15,600 cm², indicating that a growing number of contacts is formed by a declining number of presynaptic neurons. © 1992 John Wiley & Sons, Inc.     

Dual role of osteoblastic proenkephalin derived peptides in skeletal tissues

Proenkephalin encodes a group of small peptides with opiate-like activity, the endogenous opioids, known to function as neurohormones, neuromodulators, and neurotransmitters. Recently, we have demonstrated that in addition to its abundance in fetal brain tissue, proenkephalin is highly expressed in nondifferentiated mesodermal cells of developing fetuses. We identified the skeletal tissues, bone, and cartilage as major sites of proenkephalin expression. To examine the possibility that proenkephalin is involved in bone development we have studied the expression of this gene in bone-derived cells, its modulation by bone active hormones, and the effects of enkephalin-derived peptides on osteoblastic phenotype. Our studies revealed that osteoblastic cells synthesize high levels of proenkephalin mRNA which are translated, and the derived peptides are secreted. Reciprocal interrelationships between osteoblast maturation and proenkephalin expression were established. These results together with our observations demonstrating inhibitory effects of proenkephalin-derived peptides on osteoblastic alkaline phosphatase activity, strongly support the notion that proenkephalin is involved in bone development. A different direction of research by other investigators has established the capability of the opioid system in the periphery to participate in the control of pain. On the basis of these two lines of observation, we would like to present the following hypothesis: The potential of embryonic skeletal tissue to synthesize proenkephalin-derived peptides is retained in the adult in small defined undifferentiated cell populations. This potential is realized in certain situations requiring rapid growth, such as remodeling or fracture repair. We suggest that in these processes, similarly to the situation in the embryo, the undifferentiated dividing cells produce the endogenous opioids. In the adult these peptides may have a dual function, namely participating in the control of tissue regeneration and in the control of pain. © 1994 Wiley-Liss, Inc.     

Ultrastructure of the early stages of carposporophyte development in the red algaChondria tenuissima (Rhodomelaceae,Ceramiales)

The ultrastructure of the early stages of carposporophyte development in the marine red algaChondria tenuissima has been studied. The diploid carposporophyte grows on the gametophyte. Apical gonimoblast cells develop into diploid carpospores. The basal gonimoblast cells cease to divide and undergo considerable cytoplasmic changes before they become incorporated into the expanding fusion cell. Nucleus and plastids degenerate gradually, while mitochondria remain intact. The smooth endoplasmic reticulum becomes prominent, it seems to produce small vesicles with electron dense contents. Simultaneously, numerous mucilage sacs are formed, presumably from dilating ER cisternae. The contents of the mucilage sacs are secreted by exocytosis. The pit connections between gonimoblast cells flare out. They remain as isolated bodies without connection to a wall after fusion. Secondary pit connections occur between vegetative gametophyte cells and sterile carposporophyte cells. There are three different morphological types of pit connections.     

Intraspecific Variation in Cryptocaryon irritans

ABSTRACT. Intraspecific variation in the ciliate Cryptocaryon irritans was examined using sequences of the first internal transcribed spacer region (ITS-1) of ribosomal DNA (rDNA) combined with developmental and morphological characters. Amplified rDNA sequences consisting of 151 bases of the flanking 18 S and 5.8 S regions, and the entire ITS-1 region (169 or 170 bases), were determined and compared for 16 isolates of C. irritans from Australia, Israela and the USA. There was one variable base between isolates in the 18 S region nd 11 variables indicate that Australian C. irritans isolates from estuarine (Moreton Bay) and coral reef (Heron Island) environments are distinct. The Heron Island isolate was genetically closer to morphologically dissimilar isolates from Israel (1.8% divergence) and USA (2.3 % divergence) than it was to the Moreton Bay isolates. Three isolates maintained in our laboratory since February 1994 originated from the same source. During this time the sequence of the isolates from wild fish in Moreton Bay remained unchanged. These genetic differences indicate the existence of a founder effects in laboratorty populations of C. irritans . The genetic variation found here, combined with known morphological and developmental differences, is used to characterise four strains of C. irritans .     

How do the consumption and development rates of the conifer specialist Aphidecta obliterata respond to temperature,and is it better adapted to limited prey than a generalist?

This study aimed to address the effect of temperature on the consumption and development rates of Aphidecta obliterata and to compare the responses of Ap. obliterata (specialist) with that of Adalia bipunctata (generalist) to prey limitation. Temperature had a significant and positive effect on the time to egg hatch of Ap. obliterata . The duration of the larval instars was not affected by prey species at 15°C but was significantly shorter, 12.1 days at 20°C compared with 21.1 days at 15°C. The proportion of time spent in each instar, however, was not affected by temperature or prey species, but the duration of the pupal stage was significantly affected by temperature. The average daily consumption of prey aphids increased with instar and was significantly influenced by temperature. There was a significant difference in the length of the pupal stage between coccinellid species but not that of the larval stage. The duration of the larval period increased under conditions of prey shortage. The pupal period of Ap. obliterata was significantly affected by the food regime but not that of Ad. bipunctata . There was a significant interaction between species and food supply on the length of the pupal stage and the larval stage and the final fresh weight achieved by the newly emerged adults. Male adults weighed significantly less than the females in all regimes. Larvae of Ap. obliterata and Ad. bipunctata did not consume any of the alternative prey (Collembola or Psocoptera) provided. There was no significant difference in the consumption of prey between the two coccinellid species. The results suggest that both of these coccinellids are well adapted to low-density-specific prey. There were no obvious differences between the two, which would tend to favour either species in an environment of limited prey.     

Corporate sustainability for architecture engineering and construction (AEC) organizations: Framework,transition and implication strategies

This research aims to propose a regenerative sustainability framework for AEC organizations, not only to focus on delivering green certified projects, but to encourage a revitalized approach to systematically drive their sustainability initiative. To achieve this aim, the study conducts an extensive global review of sustainable assessment systems in various industries and organizations, such as corporate sustainable reports, green company competitions, and green-renowned stocks. After learning the best practices from other industries, a sustainable development framework for the AEC organizations is developed. The proposed framework constitutes 110 key indicators covering three-dimensions of sustainability—social, environmental, and economic, respectively—and four-elements of corporate development essentials—projects, operations, governance, and stakeholders. By comparing the proposed framework to existing AEC practices, the study identifies the skewed development of corporate sustainability in the AEC industry where much awareness have been given to project-level environmental matters, but the social dimension, such as social stakeholders and social governance, are seriously lacking and need to be prioritized. A case study of a multi-millions construction company in China was investigated to validate this framework in practice. The case analysis justifies the practical value of the proposed framework, and elaborates the future need of regenerative sustainable initiatives for AEC organizations. The study contributes to the development of corporate sustainability theory in the AEC industry, and also provides industrial practitioners and policy makers with a better understanding of the next generation of corporate sustainable performance and implementation strategies.     

Oxygen: a master regulator of pancreatic development?

Beyond its role as an electron acceptor in aerobic respiration, oxygen is also a key effector of many developmental events. The oxygen‐sensing machinery and the very fabric of cell identity and function have been shown to be deeply intertwined. Here we take a first look at how oxygen might lie at the crossroads of at least two of the major molecular pathways that shape pancreatic development. Based on recent evidence and a thorough review of the literature, we present a theoretical model whereby evolving oxygen tensions might choreograph to a large extent the sequence of molecular events resulting in the development of the organ. In particular, we propose that lower oxygenation prior to the expansion of the vasculature may favour HIF (hypoxia inducible factor)‐mediated activation of Notch and repression of Wnt/β‐catenin signalling, limiting endocrine cell differentiation. With the development of vasculature and improved oxygen delivery to the developing organ, HIF‐mediated support for Notch signalling may decline while the β‐catenin‐directed Wnt signalling is favoured, which would support endocrine cell differentiation and perhaps exocrine cell proliferation/differentiation.     

Dissection and Culture of Mouse Dopaminergic and Striatal Explants in Three-Dimensional Collagen Matrix Assays

Midbrain dopamine (mdDA) neurons project via the medial forebrain bundle towards several areas in the telencephalon, including the striatum¹. Reciprocally, medium spiny neurons in the striatum that give rise to the striatonigral (direct) pathway innervate the substantia nigra². The development of these axon tracts is dependent upon the combinatorial actions of a plethora of axon growth and guidance cues including molecules that are released by neurites or by (intermediate) target regions^3,4. These soluble factors can be studied in vitro by culturing mdDA and/or striatal explants in a collagen matrix which provides a three-dimensional substrate for the axons mimicking the extracellular environment. In addition, the collagen matrix allows for the formation of relatively stable gradients of proteins released by other explants or cells placed in the vicinity (e.g. see references 5 and 6). Here we describe methods for the purification of rat tail collagen, microdissection of dopaminergic and striatal explants, their culture in collagen gels and subsequent immunohistochemical and quantitative analysis. First, the brains of E14.5 mouse embryos are isolated and dopaminergic and striatal explants are microdissected. These explants are then (co)cultured in collagen gels on coverslips for 48 to 72 hours in vitro. Subsequently, axonal projections are visualized using neuronal markers (e.g. tyrosine hydroxylase, DARPP32, or βIII tubulin) and axon growth and attractive or repulsive axon responses are quantified. This neuronal preparation is a useful tool for in vitro studies of the cellular and molecular mechanisms of mesostriatal and striatonigral axon growth and guidance during development. Using this assay, it is also possible to assess other (intermediate) targets for dopaminergic and striatal axons or to test specific molecular cues.