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Partial sciatic nerve injury, a model of neuropathic pain, elicits a variety of neurochemical, electrophysiological and neuroanatomical changes in primary sensory neurons. We have used the technique of messenger RNA differential display to identify genes with altered expression in these neurons which may contribute to the development of aberrant sensation following such peripheral nerve damage. This approach identified 14 distinct complementary DNA clones, representing transcripts with increased ipsilateral expression in L4/5 dorsal root ganglia, two weeks after unilateral partial ligation of the rat sciatic nerve. Both Zucker diabetic fatty rats and their lean counterparts were used in this study but none of the transcripts identified showed an induction that was confined to one of the two groups. The majority of the clones did not show significant sequence similarity to previously reported genes and therefore may represent novel messenger RNA sequences or, alternatively, unknown regions of partially characterised messenger RNAs. Two of the clones represented transcripts for the known proteins muscle LIM protein and acidic epididymal glycoprotein, neither of which had previously been associated with expression in the nervous system. Reverse transcriptase-polymerase chain reaction analysis and in situ hybridization confirmed that the messenger RNA expression of both muscle LIM protein and acidic epididymal glycoprotein was induced in an ipsilateral-specific manner. Their localisations, examined with in situ hybridization in L5 dorsal root ganglia, were limited in each case to a sub-population of neuronal profiles. Those neuronal profiles that demonstrated muscle LIM protein hybridization were distributed across the profile size range, whereas the distribution of acidic epididymal glycoprotein-positive profiles appeared to be skewed towards smaller profiles. The induction of muscle LIM protein and acidic epididymal glycoprotein in dorsal root ganglia may play an important functional role in the adaptive response of primary sensory neurons following partial sciatic nerve injury.

Partial injury of the rat sciatic nerve elicits a variety of characteristic chemical, electrophysical and anatomical changes in primary sensory neurons and constitutes a physiologically relevant model of neuropathic pain. To elucidate molecular mechanisms that underlie the physiology of neuropathic pain, we have used messenger RNA differential display to identify genes that exhibit increased ipsilateral expression in L4/5 dorsal root ganglia, following unilateral partial ligation of the rat sciatic nerve. One set of partial complementary DNA clones identified in this screen was found to encode a protein kinase, nerve injury-associated kinase. Cloning of the full-length human nerve injury-associated kinase complementary DNA, together with recombinant expression analysis, reveal nerve injury-associated kinase to be a functional member of a subgroup of sterile 20-like protein kinases characterised by the presence of a putative carboxy terminal autoregulatory domain. Induction of nerve injury-associated kinase expression in dorsal root ganglia in the rat neuropathic pain model was confirmed by quantitative reverse transcription-polymerase chain reaction, and RNA in situ hybridization analysis revealed enhanced levels of nerve injury-associated kinase within neurons.Together, our data implicate nerve injury-associated kinase as a novel upstream component of an intracellular signalling cascade that is up-regulated in dorsal root ganglia neurons in response to sciatic nerve injury.

A quantitative light- and electron-microscopic study has been made of the retinae of rats which were exposed to different lighting conditions for between one and 15 weeks in young adulthood, having been reared in identical conditions during development. The width of the inner and outer segments of the photoreceptors and the width of the outer plexiform layer varied inversely with the light intensity under diurnal lighting conditions of 10 h light/14 h dark. Linear regression analysis showed that the widths were inversely related to the fourth root of the light intensity as measured in lux. Both central and peripheral areas of retina showed a similar change. No change was seen in the widths of the inner plexiform layer, or of the inner and outer nuclear cell layers. Nor was there a difference in the packing density or size of the nuclei in the nuclear cell layers. The number of ribbon synapses in the outer plexiform layer also varied inversely with the intensity of diurnal light. Linear regression analysis showed that the number of synapses was inversely correlated with the fourth root of the light intensity and was positively correlated with the width of the outer plexiform layer. The number of ribbon synapses was increased by up to two and a half times in constant darkness compared to diurnal light of 35 lux. The increase was present but not maximal after one week of exposure. The length of synaptic ribbons was unchanged. The nerve terminals forming such synapses were increased in size but not in number. After one week, there was little or no additional change in the retinal widths and number of synaptic ribbons with time. However, there was a progressive increase with time in nerve terminal size (two-fold in area) in constant darkness. There was some evidence of a slight decrease in nerve terminal number and increase in size of retinal nuclei with age. It is concluded that the adult retina responds to a different lighting environment by a relatively rapid change in the size of photoreceptor segments, by a progressive and large change in number of ribbon synapses and by a slower progressive and large change in the size of photoreceptor nerve terminals. The response is quantitatively determined by the strength of the stimulus but not in a linear fashion. These results are compared with the effects of environmental stimulation of other areas of the nervous system.

Groups and sub-groups (clusters) of small granule-containing cells ("small cells") were analysed at 3 and 6 micron intervals and in serial sections, in rats aged 2-13 months. Fully intraganglionic clusters of small cells were all found to receive an incoming ("afferent") innervation, of the order of 3-6 afferent terminals per cell, derived from axons of preganglionic type via multifocal, symmetrical, mainly axosomatic synapses. No evidence was obtained of sharing of preganglionic inputs between small cells and principal neurones. Intraganglionic clusters also regularly gave outgoing ("efferent") synapses of the asymmetrical type, of the order of 2-6 per cell, to intraganglionic nerve elements; 30-50% of these synapses were given from somata, 50-70% from processes of the small cells. Whenever the postsynaptic structure was identifiable these synapses were all found to be given to postganglionic neurones or their dendrites, principally to spine-like processes or slender twigs. In some ganglia a few efferent synapses to other small cells were observed; these were of the symmetrical type. Efferent synapses to nerve profiles resembling chemosensory axon terminals, also of the symmetrical type, were extremely infrequent (fewer than 1% of all efferent synapses) in intraganglionic small cell groups and appeared virtually restricted to glomus-like clusters of small cell, which lay intracapsularly, or in and near the bases of nerves entering or leaving the ganglion. Almost all groups and clusters of small cells were located near to fenestrated capillary vessels, which are not found elsewhere in the ganglion. The implications of possible non-synaptic release of material from small cells via membrane regions not covered by satellite cell cytoplasm, were explored in a nearest-neighbour analysis. These "exposed" regions comprised 1-3% of the small cell surface, a proportion comparable with those engaged in receiving afferent synapses or in giving efferent synapses. The majority of such regions faced toward other nerve profiles (axons and dendrites) ensheathed in satellite cytoplasm (mean 30%), intraganglionic tissue spaces wider than 3 micron (mean, 30%) or other small cells (mean, 14%); 25% faced toward blood vessels, but of these vascularly directed regions, only one fifth (or 5% of the total) on average faced directly toward fenestrated endothelium, the rest being non-fenestrated and/or separated by pericyte processes from the exposed regions of small cell membrane. Thirty-three percent of the small cells in a sample of 242 lay within 2 micron of the nearest blood vessel.(ABSTRACT TRUNCATED AT 400 WORDS)