Ontogenetic changes in the regenerative ability of chick retinal ganglion cells as revealed by organ explants

Summary Whereas mature neurons in the central nervous system (CNS) of birds lack the capability of regenerating axons after injury, embryonic nerve cells are able to do so. The time course of this decline of regenerative ability was investigated in ganglion cells from embryonic chick retinae. Retinal strips from 7-to 19-day-old embryos (E7–E19) were explanted and cultured in vitro. The numbers of retinal ganglion cell (RGC) neurites that had extended during the first 22–23 h incubation, their elongation rates, and morphometric parameters of the growth cones were measured to characterize the regenerative behavior. We observed two periods of decline in neuritic growth: the first from E7 to E9, and another from E14 to E19. The first decrease may reflect a gradually disappearing portion of neurons which produced their axons de novo. The second decline coincides with the major period of synaptogenesis by ganglion cell axons in ovo. The time required for initiation of axonal outgrowth increased, accordingly, from less than 3 h in explants from younger retinae (E7–E16) to 10–12 h for E17 and E19 explants. Axonal elongation rates ranged between 36 m/h and 56 m/h (mean values) for E7–E13 explants, but were significantly lower for cells from E14–E19 retinae (13–21 m/h). Morphologically, neurites and growth cones for RGC explanted before E17 were characterized by their high variability. They possessed more filopodia than mature neurons (E17, E19), fasciculated to a higher degree and branched more frequently. In addition, older neurites appeared stiffer and were morphologically simpler. We propose that, during the embryogenesis of the chick retinofugal system, the changes that render mature ganglion cells incapable of axonal regeneration are mainly triggered in the period of establishing synaptic contacts in the optic tectum.     

Phytochrome control of skotodormancy release in Grand Rapids lettuce achenes

Light-requiring Grand Rapids lettuce ( Lactuca sativa L.) achenes develop skotodormancy when imbibed in darkness for 7 days at 25°C. Redried skotodormant achenes maintain this type of dormancy upon subsequent rehydration. At 25°C full germination of skotodormant achenes can be induced by continuous and intermittent red light illumination as well as by several brief red irradiations given daily. One brief (10 min) red light irradiation can partly break skotodormancy at 20°C, while at lower temperatures the same treatment results in full induction of germination. Phytochrome control of the release from skotodormancy is proven by a) the dependence of the germination response on the relative sequence of red and far-red light in cyclic irradiations, and b) the reversion of red action by subsequent far-red irradiation. The time course of germination of skotodormant achenes treated with intermittent red light depends upon the length of dark interval between the light pulses. Germination is considerably delayed compared to that of non-skotodormant ones, induced by a single brief red light treatment. This fact in combination with the requirement, over a long period of time, of P_fr action for full manifestation of germination, indicates that skotodormancy is a deeper form of dormancy. It is concluded that the germination of lettuce achenes may always be subjected to phytochrome control.     

Coactivation of pre- and postsynaptic signaling mechanisms determines cell-specific spike-timing-dependent plasticity

Synapses may undergo long-term increases or decreases in synaptic strength dependent on critical differences in the timing between pre-and postsynaptic activity. Such spike-timing-dependent plasticity (STDP) follows rules that govern how patterns of neural activity induce changes in synaptic strength. Synaptic plasticity in the dorsal cochlear nucleus (DCN) follows Hebbian and anti-Hebbian patterns in a cell-specific manner. Here we show that these opposing responses to synaptic activity result from differential expression of two signaling pathways. Ca2+/calmodulin-dependent protein kinase II (CaMKII) signaling underlies Hebbian postsynaptic LTP in principal cells. By contrast, in interneurons, a temporally precise anti-Hebbian synaptic spike-timing rule results from the combined effects of postsynaptic CaMKII-dependent LTP and endocannabinoid-dependent presynaptic LTD. Cell specificity in the circuit arises from selective targeting of presynaptic CB1 receptors in different axonal terminals. Hence, pre- and postsynaptic sites of expression determine both the sign and timing requirements of long-term plasticity in interneurons.     

Topographic representation of the sciatic nerve motor neurons in the spinal cord of the adult rat correlates to region-specific activation patterns of microglia

The frequent use of the adult rat sciatic nerve as a model to study the neuronal responses to injury, nerve regeneration and in transplantation studies, requires a detailed knowledge of the projection pattern of motor neurons into this nerve. Thus, as a first goal we determined this topographical projection of motor neurons and labelled small contingents by applying the fluorescent dye DiI in localised incisions made in the dorsal, rostral, ventral or caudal quadrants of the nerve. As a second goal we analysed with immunohistochemical methods the response of microglial cells within the topographical area corresponding to the incision and within areas outside this location. Uptake of the dye occurred only within the area confined to the incision, thus allowing the identification of the corresponding motor neuron perikarya within the ventral horn, eight to ten days later. In serial transverse sections of the lumbosacral spinal cord the number of labelled cells, their position within the ventral horn, and their longitudinal extent have been determined. The data suggest that the gross projection of the lumbosacral motor neuron column at the mid-thigh level of the sciatic nerve is topographic. In accordance, microglial cells showed fast activation within the injured topographic area, and a less pronounced and delayed response within the non-injured areas of the ventral horn. The graded response of microglial cells suggests that these cells possess a potential of local activation by sensing whether neurons are axotomised or just irritated by axotomy of their neighbours. The topographic organisation proves to be useful in studies on local injuries to the sciatic nerve and when analysing retrograde responses within the lumbosacral spinal cord.     

53BP1, an activator of ATM in response to DNA damage

p53 Binding protein 1 (53BP1) belongs to a family of evolutionarily conserved DNA damage checkpoint proteins with C-terminal BRCT domains and is most likely the human ortholog of the budding yeast Rad9 protein, the first cell cycle checkpoint protein to be described. 53BP1 localizes rapidly to sites of DNA double strand breaks (DSBs) and its initial recruitment to these sites has not been shown to be dependent on any other protein. Initially, 53BP1 was thought to be a mediator of DNA DSB signaling, but now it has been shown to function upstream of ataxia-telangiectasia mutated (ATM), in one of at least two parallel pathways leading to ATM activation in response to DNA damage. Currently, only a single tudor and two BRCT domains are recognized in 53BP1; however, their precise functional role is not understood. Elucidating the function of 53BP1 will be critical to understanding how cells recognize DNA DSBs and how ATM is activated.     

Photoinhibition of Seed Germination in the Maritime Plant Matthiola tricuspidata

The three-horned stock, Matthiola tricuspidata (L.) R. Br. isa widespread annual plant of the Mediterranean sandy shores.Its seeds are dark germinating and negatively photosensitive,in accordance with our previous findings for a number of othermaritime plants. Full germination was obtained at a wide rangeof temperatures (5-25 °C) in the dark. Inhibition of germinationunder light of various spectral qualities could be generallycorrelated, negatively and positively, respectively, with phytochromephotostationary state (ø) and relative cycling rate ofphytochrome (H). The inhibition of germination by white (fluorescent),blue and far-red light, applied either continuously or intermittently,consistently showed a linear dependence upon the logarithm ofthe flux density of the irradiation. The resulting photoinhibitioncurves had parallel slopes and, compared to those of other maritimeplants, they were shifted to higher flux densities, Continuousblue or far-red irradiations, both establishing a similar øvalue (0·26), resulted in statistically similar regressioncurves, thus favouring the hypothesis that phytochrome is thesingle photoreceptor in the photoinhibition of seed germination.Copyright1994, 1999 Academic Press Matthiola tricuspidata (L.) R. Br., three-horned stock, seed germination, light, photoinhibition, phytochrome     

Characterization of polymorphic microsatellite markers for the endangered Mediterranean bath sponge Spongia officinalis L.

Ten polymorphic microsatellite markers are described for the Mediterranean bath sponge Spongia officinalis. Loci were isolated from a genomic library enriched for AC repeats. Microsatellite markers were evaluated on a Cretan population of 28 individuals; the allelic richness ranged from 5 to 34 with an average of 17, while expected and observed heterozygosities varied from 0.505 to 0.964 and 0.444 to 0.963, respectively. In a species whose populations in the eastern Mediterranean basin have been substantially declined due to recurring mass mortality incidents and intensive harvesting, these markers are expected to assist studies of genetic structure and differentiation between populations.