It has been well established that axotomized retinal ganglion cells (RGCs) of adult mammals can survive and regenerate their axons when the peripheral nerve is autografted [1]. However, the number of RGCs with regenerated axons is still limited. One major reason for poor regeneration is that many RGCs degenerate shortly after optic nerve (ON) transection. For example, half of the rat RGCs were lost within 1 week after intraorbital ON transection, and only 10% survived 2 weeks after axotomy [2]. The rescue of axotomized RGCs from such early retrograde degeneration is a prerequisite for the functional recovery of vision against ON damage.

Optic nerve (ON) injury in adult mammals causes retinal ganglion cell (RGC) death and subsequent visual loss. Recovery of vision requires both rescuing axotomized RGCs and inducing their axonal regeneration. Axotomized RGCs are significantly rescued by overexpression of bcl-2, an anti-apoptotic gene. However, whether bcl-2 affects axonal regeneration is controversial. In neonatal bcl-2 transgenic mice (bcl-2 mice), optic tract regeneration after tectal lesion was promoted (Chen et al., 1997), whereas ON regeneration after ON crush was not (Lodovichi et al., 2001). These conflicting results may be attributable to different environments between tectum and ON. We tested here whether bcl-2 overexpression enhances in vivo RGC axonal regeneration in adult mice through a permissive environment in the peripheral nerve (PN) graft. Four weeks after PN transplantation to the proximal ON stump, we assessed the number of surviving and regenerating RGCs by retrograde labeling. Although the survival rate in bcl-2 mice was significantly enhanced compared with that in wild-type (wt) mice, the regeneration rate was not enhanced. In both bcl-2 and wt mice, RT97 immunostaining of the PN-grafted retinas revealed some RGC axons regrowing intraretinally but repulsed at the optic disk. To circumvent this repulsive barrier, we directly transplanted the PN graft to the partially injured retina and compared regeneration rates between these mice. Here again the regeneration rate in bcl-2 mice did not exceed that in wt mice. These findings indicate that bcl-2 overexpression enhances survival but not axonal regeneration of adult RGCs even within a permissive environment.

Purpose: Retinal ganglion cells (RGCs) of adult mammals can regenerate their axons along a segment of the peripheral nerve (PN) that is transplanted to the cut optic nerve. There have been many trials of PN transplantation to induce axonal regeneration of RGCs in adult rodents, cats and ferrets. However, because of the technical difficulty in transplant operation, PN transplantation in adult mice has not been carried out in spite of the availability of many kinds of gene-manipulated animals. Here we report the procedures for successful PN transplantation in this species. Methods: We made intraretinal (IR) and retrobulbar (RB) approaches for PN transplantation. Four weeks after PN transplantation, RGCs with regenerated axons were identified by retrograde labeling with rhodamine or horseradish peroxidase applied into the PN segment. Results: A quantitative survey showed that the mean regeneration ratio was 1.0 % (n = 8) in IR transplantation, whereas it was only 0.1 % in RB transplantation (n = 11). As previously shown in other species, the regenerated RGCs were predominantly larger-bodied cells in com-parison to intact cells. Conclusion: Possible reasons for the difference in regeneration ratio between the two transplant approaches and the feature of soma size of regenerated RGCs are discussed.

A visual prepulse was applied just before a burst of white noise with various lead intervals ranging from 50 to 400 ms in hamsters. After habituation each hamster received 120 trials of the auditory noise with or without the prepulse. Amplitude of startle response increased significantly at 100 ms and 200 ms (n = 10). This enhancement disappeared in blind controls (n = 6). The amplitude of startle response decreased during exploratory behavior, grooming, sniffing or other movements. However, the facilitatory effects of the visual prepulse were consistent, regardless of the animal's behavioral states, except sniffing. The authors conclude that visual prepulse task is useful for studying the ability to detect visual signals in hamsters, and that further study is necessary to elucidate the factors affecting the enhancement of startle response in hamsters.

Covalent organic framework TpBD was grafted on stainless steel wire with polydopamine as a linker. The fabricated TpBD bonded stainless steel wire was used as the solid-phase microextraction fiber to extract sixteen polycyclic aromatic hydrocarbons (PAHs) for subsequent GC-MS/MS determination in grilled meat samples. The developed method gave the limits of detection (S/N = 3) from 0.02 (pyrene)-1.66 (naphthalene) ng L−1 and enhancement factors from 1069 (naphthalene)-10879 (benz(a)anthracene). The relative standard deviations (RSDs) for intra-day and inter-day study are in the range of 2.6%–8.5% and 4.5%–9.4%, respectively. The fiber-to-fiber RSDs for three parallel prepared fibers were 5.3%–10.0%. One TpBD bonded fiber can stand at least 200 cycles without significant loss of extraction efficiency. The developed method was successfully applied for the determination of trace PAHs in grilled meat samples with recoveries from 85.1% to 102.8%.

In a previous study, we showed that the visual ON response recorded from the superior colliculus is absent in the metabotropic glutamate receptor subtype 6 (mGluR6) knockout mice. However, these mutant mice can respond to some visual inputs and are able to learn a light-dark discrimination task. Here, we systematically investigated the possibility that some ON responses could remain in the mutant mice. Recording light evoked field potentials from the superior colliculus of mGluR6-deficient mice at scotopic and mesopic backgrounds, we found a characteristic ON response that differs from the ON response of wild-type mice. The ON responses recorded from the mutant mice appeared with a latency (200–300 ms) longer than that of the wild-type mice (50–100 ms) and amplitudes of this late ON response decreased upon increase of the light intensity in most cases examined. The late ON response may contribute to the apparent normal behavior and some physiological responses to light stimuli in mGluR6-deficient mice.

Primary Sjögren’s syndrome (pSS) is an autoimmune disease that targets salivary and lachrymal glands, characterized by anti-cholinergic autoantibodies directed against the M3 muscarinic acetylcholine receptor (mAChR). The aim of this work was to evaluate if cholinergic autoantibodies contained in IgG purified from Sjögren sera could trigger apoptosis of A253 cell line. We also determined if caspase-3 and matrix metalloproteinase-3 (MMP-3) are involved in the induction of A253 cell death. Our results demonstrated that anti-cholinergic autoantibodies stimulate apoptosis and inositol phosphate (InsP) accumulation accompanied by caspase-3 activation and MMP-3 production. All of these effects were blunted by atropine and J104794, indicating that M3 mAChRs are impacted by the anti-cholinergic autoantibodies. The intracellular pathway leading to autoantibody-induced biological effects involves phospholipase C (PLC), calcium/calmodulin (CaM) and extracellular calcium as demonstrated by treatment with U-73122, W-7, verapamil, BAPTA and BAPTA-AM, all of which blocked the effects of the anti-cholinergic autoantibodies. In conclusion, anti-cholinergic autoantibodies in IgG purified from pSS patient’s sera mediates apoptosis of the A253 cell line in an InsP, caspase-3 and MMP-3 dependent manner.

The auditory brainstem response and histopathology of the cochlea were investigated in an accelerated senescence-prone strain, SAM-P/1 mice and a senescence-resistant strain, SAM-R/1 mice. Each strain displayed an age-related auditory loss expressed as elevated thresholds similar to human hearing loss in that high-frequency losses occurred earlier than middle- or low-frequency losses. SAM-P/1 showed a more rapid decline of hearing with age than did SAM-R/1. Interpeak intervals I–III and I–IV were prolonged with age in both strains, especially at high frequency. The prolongation was more marked in SAM-P/1 than in SAM-R/1. The decrease in amplitude of wave I observed in both strains was greater in SAM-P/1 than in SAM-R/1. The auditory function assessed by thresholds, interpeak intervals and amplitudes of wave I in SAM-P/1 at 12 months of age corresponded roughly to that in SAM-R/1 at 20 months of age. In morphological studies, there was an age-related decrease in the cell density as well as in the size of spiral ganglion neurons in both strains, but these changes were more pronounced in SAM-P/1 than in SAM-R/1. These results reveal that age-related hearing impairment associated with morphological changes in the cochlea is manifested earlier and progresses more rapidly in SAM-P/1 than in SAM-R/1. Thus, the SAM-P/1 strain should prove useful as a model of presbycusis.

In adult golden hamsters (Mesocricetus auratus), the optic nerve was bilaterally sectioned with one side grafted with the sciatic nerve to make a bridge to the ipsilateral superior colliculus. Shuttle-box avoidance was tested using light as the conditioned stimulus. Three of the five transplanted animals revealed statistically significant increase in percentages of avoidance. A significant increase in the avoidance scores was also observed in 15 normal hamsters, but none of 13 blind hamsters showed such an increase. Intertrial responses, which represent spontaneous responses, did not show significant changes. We conclude that some of the transplanted animals can discriminate between light and dark, probably through their restored visual pathway.

We studied the morphology of regenerated retinal ganglion cells and their axons in adult rodents after axotomy and autologous transplantation of the sciatic nerve. Regenerated ganglion cells, backlabeled with rhodamine dextran, were of similar size to or larger than those of intact cells in control animals. Dendrites and occasionally axons as well showed abnormal morphologies in most cells, though some cells appeared quite normal. Cross-sections of the regenerated axons, observed by electron microscopy, were always attached to either the Schwann cell cytoplasm or the basal lamina. The immunoreactive structures to anti-laminin antibody were quite irregular in the cross-sectioned graft and, compared with those of the intact sciatic nerve, they were generally smaller. Their appearance closely resembled that of the basal lamina in the graft observed by electron microscopy. These observations, taken together, suggest that the laminin-rich basal laminae of Schwann cells are essentially important for the regeneration of retinal axons in adult rodents.