Quantitative changes in mitochondria of spinal ganglion neurons in aged rabbits

Within the context of our research on the age-related structural changes in spinal ganglia, we studied the mitochondria of the neuronal perikaryon in the spinal ganglia of 12-, 42-, and 79-month-old rabbits. Both the volume of the perikaryon and the total mitochondrial mass within the perikaryon increased significantly passing from young adult to old animals. Hence, there is no net loss of mitochondria in these neurons with age. Since, however, the volume of the perikaryon increased by more than 63% while the total mitochondrial mass within the perikaryon increased by only 18%, the mean percentage of perikaryal volume occupied by mitochondria decreased with age. This decrease is only in very minor part a consequence of lipofuscin accumulation, so that the ratio between the total mitochondrial mass and the functionally active volume of cytoplasm decreased with age. Possible causes of this decrease are discussed briefly. Moreover, while the mitochondrial structure did not change, mitochondrial size increased with age. Finally, in each of the three age groups both the mean percentage volume of mitochondria and the mean mitochondrial size were very similar in large light and in small dark neurons. Copyrigh

A study of mitochondria in spinal ganglion neurons during life : quantitative changes from youth to extremely advanced age

In view of the central role that mitochondria are thought to play in the ageing process, we investigated changes in mitochondria of spinal ganglion neurons in rabbits aged 1, 3.6, 6.7, and 8.8 years (the latter extremely old). Mitochondrial size increased significantly with age, while mitochondrial structure did not change. The total volume of mitochondria within the perikaryon did not change significantly during life. This indicates that in these neurons mitochondrial degradation was completely compensated by the production of new mitochondria even in the extremely advanced age. We also found that the mean volume of neuronal perikaryon increased markedly with age, so that the mean percentage of perikaryal volume occupied by mitochondria decreased significantly with a difference of about 33% between the youngest and the oldest animals. This decrease is only in very small part due to lipofuscin accumulation, so that the ratio of the total volume of mitochondria to the volume of functionally active cytoplasm decreased with age. The mitochondria of the neurons studied seem therefore unable to adapt their total volume to the volume of functionally active cytoplasm, that increases with age. This result is consistent with the observation that the neurons of old animals have a reduced ability to respond to high energy demands

Age-related quantitative changes in mitochondria of satellite cell sheaths enveloping spinal ganglion neurons in the rabbit

We studied mitochondria in the satellite cell sheaths which envelope the spinal ganglion neurons of rabbits aged 12, 42, and 79 months. While the mean cytoplasmic volume of satellite cell sheaths did not change significantly with age, the mean percentage of cytoplasmic volume occupied by mitochondria decreased with age. This decrease is mainly due to a reduction in the total mitochondrial mass and only in minor part is a consequence of lipofuscin accumulation. Mitochondrial structure did not change, while mitochondrial size increased with age. Comparison between mitochondria in nerve cell bodies and those in satellite cell sheaths showed that: (1) the mean percentage of cytoplasmic volume occupied by mitochondria was greater in nerve cell bodies than satellite cell sheaths and the ratio between these two percentages remained constant with advancing age; (2) the total mitochondrial mass was much greater in nerve cell bodies than satellite cell sheaths and the ratio between these two values increased with age; (3) the extent of increase of mitochondrial size with age was similar in nerve cell bodies and satellite cell sheaths. The results of the present study suggest that: (1) the ability of satellite cell sheaths to produce energy decreases with age; (2) the decreased ability of sensory neurons in old animals to meet high energy demands may be partly due to the diminished contribution of their associated satellite cell sheaths

The perineuronal glial tissue of spinal ganglia : quantitative changes in the rabbit from youth to extremely advanced age

The volumes of the nerve cell bodies and those of the enveloping satellite cell sheaths from spinal ganglia were determined by morphometric methods applied to electron micrographs in young, adult, old and very old rabbits. The mean volume of the nerve cell bodies increased progressively with age; this is probably related to the increase with age of the body size of the rabbits studied. The mean volume of the satellite cell sheaths did not differ significantly in young, adult and old animals, but was significantly smaller in very old animals. It is extremely unlikely that this marked reduction in the volume of the satellite cell sheath is the result of a pathological process. The mean value of the volume ratio between the satellite cell sheaths and the related nerve cell bodies did not differ significantly in young and adult animals, but was significantly smaller in old and very old animals. This ratio was particularly low in very old animals. Our analysis showed that in each age group the volume of the satellite cell sheath is linearly related to the volume of the related nerve cell body. This result suggests that in rabbit spinal ganglia the quantitative relations between glial and nervous tissue are tightly controlled throughout life. It is suggested that ganglionic neurons release signals to influence and control the volume of their associated glial tissue. Since satellite cells have important support roles for the neurons they surround, it is likely that the marked reduction in the volume of perineuronal sheaths in the extremely advanced age is accompanied by a reduction of those roles, with negative consequences for neuronal activity

Increase in number of the gap junctions between satellite neuroglial cells during lifetime: An ultrastructural study in rabbit spinal ganglia from youth to extremely advanced age

This study investigated quantitative aspects of the gap junctions between satellite neuroglial cells that envelope the spinal ganglion neurons in rabbits aged 1 year (young), 3.6 years (adult), 6.7 years (old), and 8.8 years (very old). Both the total number of gap junctions present in 30,000 mu m(2) of surface area occupied by perineuronal satellite cells, and the density of these junctions increased throughout life, including the extremely advanced age. By contrast, the mean length of individual gap junctions did not change with age. Thus, the junctional system which provides morphological support for the metabolic cooperation between satellite cells in rabbit spinal ganglia becomes more extensive as the age of the animal increases. These results support the hypothesis that the gap junctions between perineuronal satellite cells are involved in the spatial buffering of extracellular K(+) and in neuroprotection

Caspase-2 is upregulated after sciatic nerve transection and its inhibition protects dorsal root ganglion neurons from Apoptosis after serum withdrawal

Sciatic nerve (SN) transection-induced apoptosis of dorsal root ganglion neurons (DRGN) is one factor determining the efficacy of peripheral axonal regeneration and the return of sensation. Here, we tested the hypothesis that caspase-2(CASP2) orchestrates apoptosis of axotomised DRGN both in vivo and in vitro by disrupting the local neurotrophic supply to DRGN. We observed significantly elevated levels of cleaved CASP2 (C-CASP2), compared to cleaved caspase-3 (C-CASP3), within TUNEL+DRGN and DRG glia (satellite and Schwann cells) after SN transection. A serum withdrawal cell culture model, which induced 40% apoptotic death in DRGN and 60% in glia, was used to model DRGN loss after neurotrophic factor withdrawal. Elevated C-CASP2 and TUNEL were observed in both DRGN and DRG glia, with C-CASP2 localisation shifting from the cytosol to the nucleus, a required step for induction of direct CASP2-mediated apoptosis. Furthermore, siRNAmediated downregulation of CASP2 protected 50% of DRGN from apoptosis after serum withdrawal, while downregulation of CASP3 had no effect on DRGN or DRG glia survival. We conclude that CASP2 orchestrates the death of SN-axotomised DRGN directly and also indirectly through loss of DRG glia and their local neurotrophic factor support. Accordingly, inhibiting CASP2 expression is a potential therapy for improving both the SN regeneration response and peripheral sensory recovery

Molecular Characterization of the Gastrula in the Turtle Emys orbicularis: An Evolutionary Perspective on Gastrulation

Due to the presence of a blastopore as in amphibians, the turtle has been suggested to exemplify a transition form from an amphibian- to an avian-type gastrulation pattern. In order to test this hypothesis and gain insight into the emergence of the unique characteristics of amniotes during gastrulation, we have performed the first molecular characterization of the gastrula in a reptile, the turtle Emys orbicularis. The study of Brachyury, Lim1, Otx2 and Otx5 expression patterns points to a highly conserved dynamic of expression with amniote model organisms and makes it possible to identify the site of mesoderm internalization, which is a long-standing issue in reptiles. Analysis of Brachyury expression also highlights the presence of two distinct phases, less easily recognizable in model organisms and respectively characterized by an early ring-shaped and a later bilateral symmetrical territory. Systematic comparisons with tetrapod model organisms lead to new insights into the relationships of the blastopore/blastoporal plate system shared by all reptiles, with the blastopore of amphibians and the primitive streak of birds and mammals. The biphasic Brachyury expression pattern is also consistent with recent models of emergence of bilateral symmetry, which raises the question of its evolutionary significance

Quantum information processes in protein microtubules of brain neurons

We study biologically ‘orchestrated’ coherent quantum processes in collections of protein microtubules of brain neurons, which correlate with, and regulate, neuronal synaptic and membrane activity. In this situation the continuous Schrodinger evolution of each such process terminates in accordance with the specific Diosi-Penrose (DP) scheme of ‘objective reduction’ (‘OR’) of the quantum state. This orchestrated OR activity (‘Orch OR’) is taken to result in moments of conscious awareness and/or choice. We analyze Orch OR in light of advances and developments in quantum physics, computational neuroscience and quantum biology. Much attention is also devoted to the ‘beat frequencies’ of faster microtubule vibrations as a possible source of the observed electroencephalographic (‘EEG’) correlates of consciousness

Primary Postnatal Dorsal Root Ganglion Culture from Conventionally Slaughtered Calves

Neurological disorders in ruminants have an important impact on veterinary health, but very few host-specific in vitro models have been established to study diseases affecting the nervous system. Here we describe a primary neuronal dorsal root ganglia (DRG) culture derived from calves after being conventionally slaughtered for food consumption. The study focuses on the in vitro characterization of bovine DRG cell populations by immunofluorescence analysis. The effects of various growth factors on neuron viability, neurite outgrowth and arborisation were evaluated by morphological analysis. Bovine DRG neurons are able to survive for more than 4 weeks in culture. GF supplementation is not required for neuronal survival and neurite outgrowth. However, exogenously added growth factors promote neurite outgrowth. DRG cultures from regularly slaughtered calves represent a promising and sustainable host specific model for the investigation of pain and neurological diseases in bovines