Growth of axons through a lesion in the intact CNS of fetal rat maintained in long-term culture.

The ability of neurons in the central nervous system (CNS) to grow through a lesion and restore conduction has been analysed in developing spinal cord in vitro. The preparation consists of the entire CNS of embryonic rat, isolated and maintained in culture. Conduction of electrical activity and normal morphological appearance (light microscopical and electron microscopical) were maintained in the spinal cord of such preparations for up to 7 d in culture. A complete transverse crush of the spinal cord abolished all conduction for 2 d. After 3-5 d, clear recovery had occurred: electrical conduction across the crush was comparable with that in uninjured preparations. Furthermore, the spinal cord had largely regained its gross normal appearance at the crush site. Axons stained in vivo by carbocyanine dyes had, by 5 d, grown in profusion through the lesion and several millimetres beyond it. These experiments, like those made in neonatal opossum (Treherne et al. 1992) demonstrate that central neurons of immature mammals, unlike those in adults, can respond to injury by rapid and extensive outgrowth of nerve fibres in the absence of peripheral nerve bridges or antibodies that neutralize inhibitory factors. However, unlike the opossum, in which outgrowth occurred at 24 degrees C, although there was prolonged survival of rat spinal cords at this temperature, outgrowth of axons across the lesion required a temperature of 29 degrees C. With rapid and reliable regeneration in vitro it becomes practicable to assay the effects of molecules that promote or inhibit restoration of functional connections.     

Mitochondrial bioenergetics and neuronal survival modelled in primary neuronal culture and isolated nerve terminals

Journal of Bioenergetics and Biomembranes - Mitochondria play multiple roles in the maintenance of neuronal function under physiological and pathological conditions. In addition to ATP generation,...     

Release of Glutamate, Aspartate, and γ-Aminobutyric Acid from Isolated Nerve Terminals

Abstract: With the advent of cloning, sequencing, and patchclamping techniques, knowledge of the postsynaptic actions of amino acid neurotransmitters has undergone a dramatic advance. The primary sequences of the inhibitory receptors for γ-aminobutyric acid (GABA) (Schofield et al., 1987) and glycine (Grenningloh et al., 1987) are now established, and patch-clamp analysis has elucidated many of the factors that regulate the opening of their ion channels. The excitatory glutamate receptors are being extensively characterized at both the pharmacological (reviewed by Foster and Fagg, 1984) and the electrophysiological (reviewed by Cull-Candy and Usowicz, 1987) level. In this climate, it is perhaps surprising that the fundamental presynaptic release mechanism for the amino acid neurotransmitters remains controversial.     

Intracellular Recording,Sensory Field Mapping,and Culturing Identified Neurons in the Leech,Hirudo medicinalis

The freshwater leech, Hirudo medicinalis, is a versatile model organism that has been used to address scientific questions in the fields of neurophysiology, neuroethology, and developmental biology. The goal of this report is to consolidate experimental techniques from the leech system into a single article that will be of use to physiologists with expertise in other nervous system preparations, or to biology students with little or no electrophysiology experience. We demonstrate how to dissect the leech for recording intracellularly from identified neural circuits in the ganglion. Next we show how individual cells of known function can be removed from the ganglion to be cultured in a Petri dish, and how to record from those neurons in culture. Then we demonstrate how to prepare a patch of innervated skin to be used for mapping sensory or motor fields. These leech preparations are still widely used to address basic electrical properties of neural networks, behavior, synaptogenesis, and development. They are also an appropriate training module for neuroscience or physiology teaching laboratories.     

Evidence for two different types of insecticidal P2 toxins with dual specificity in Bacillus thuringiensis subspecies.

Analysis of polypeptides in the crystalline delta-endotoxins from different Bacillus thuringiensis strains revealed two antigenically similar forms of the P2 protein which differed in molecular mass, peptide profile, and amino acid sequence. Purified preparations of the two forms displayed the characteristic dual toxicity of the P2 protein towards members of the orders Lepidoptera and Diptera in vivo but differed markedly in potency for the insects tested. Both species of the P2 protoxin, solubilized and activated by sequential proteolysis with insect gut extract and alpha-chymotrypsin, retained activity in vivo and in vitro, despite the removal of 144 residues from the N terminus. For the low-molecular-mass form, the dual insecticidal activity was reproducible in the in vitro assays.