Mouse Cervical Spinal Cord Research Articles

Progressive changes in neurofilament proteins and growth-associated protein-43 immunoreactivities at the site of cervical spinal cord compression in spinal hyperostotic mice.

Immunohistochemical examination of the expression and localization of neurofilament (NF) proteins and growth-associated protein (GAP)-43 in spinal hyperostotic (twy/twy) mice with progressive compression of the cervical spinal cord. To determine the biologic functions of NF proteins and GAP-43 in the mouse cervical spinal cord during chronic mechanical compression. The pathologic and repair process in the chronically compressed spinal cord are understood poorly. The present authors hypothesized that there existed an increased expression of NF proteins and GAP-43 in twy/twy mice during the lengthy period of spinal cord compression, which resembles compressive myelopathy. The cervical spinal cords of 54 twy mice (aged 8 weeks [n = 18], 14 weeks [n = 18], and 20 weeks [n = 18]) and 18 control animals were examined histologically. Using appropriate antibodies, sections were also stained immunohistochemically for NF proteins and GAP-43. Separation of the myelin sheath from the axon and axonal swelling with deformation were detected in the anterior and lateral funiculi of the spinalcords of 20-week-old twy/twy mice. No such changes were noted in 8-week-old twy mice. In twy/twy mice aged 8 and 14 weeks with mild-to-moderate compression, weak immunoreactivities (mainly in the white matter) for NF proteins and GAP-43 were noted; however, in 20-week-old twy/twy mice, these axons stained strongly positive and immunoreactive swollen axons were present. The relative area of GAP-43 immunoreactive axons gradually increased between 8 and 20 weeks in each column, particularly in the anterior and lateral funiculi in the contralateral side of compression. The results showed that the expression of NF proteins and GAP-43 in the white matter increased proportionally with the magnitude of spinal cord compression, and indicated the possible involvement of GAP-43 in both axonal degeneration and repair processes in the chronically compressed spinal cord.

Dorsal Spinal Cord Inhibits Oligodendrocyte Development

Oligodendrocytes are the myelinating cells of the mammalian central nervous system. In the mouse spinal cord, oligodendrocytes are generated from strictly restricted regions of the ventral ventricular zone. To investigate how they originate from these specific regions, we used an explant culture system of the E12 mouse cervical spinal cord and hindbrain. In this culture system O4+ cells were first detected along the ventral midline of the explant and were subsequently expanded to the dorsal region similar to in vivo. When we cultured the ventral and dorsal spinal cords separately, a robust increase in the number of O4+ cells was observed in the ventral fragment. The number of both progenitor cells and mature cells also increased in the ventral fragment. This phenomenon suggests the presence of inhibitory factor for oligodendrocyte development from dorsal spinal cord. BMP4, a strong candidate for this factor that is secreted from the dorsal spinal cord, did not affect oligodendrocyte development. Previous studies demonstrated that signals from the notochord and ventral spinal cord, such as sonic hedgehog and neuregulin, promote the ventral region-specific development of oligodendrocytes. Our present study demonstrates that the dorsal spinal cord negatively regulates oligodendrocyte development.

Changes in receptor levels for thyrotropin releasing hormone, serotonin, and substance P in cervical spinal cord of Wobbler mouse: a quantitative autoradiography study during early and late stages of the motoneuron disease

Receptor levels for thyrotropin releasing hormone (TRH) measured by quantitative autoradiography in the Wobbler mouse cervical spinal cord show receptor losses that may relate to the inherited loss of motoneurons, most pronounced late (at Stage 4) in the motoneuron disease. An age-related decrease of TRH and serotonin (5-HT) receptors can be seen in the ventral horn of the control specimens (normal phenotype littermate and wild-type alike). However, this pattern is missing for substance P (SP) receptors from the wild-type specimens. Therefore the age-related decrease of SP receptors detected in the Wobbler mouse strain may identify a strain-related defect in SP neuronal/receptor developmental patterns. A higher level of TRH receptors was measured in the Wobbler dorsal horn at an early stage (Stage 1) in the motoneuron disease compared with the control specimens. The data are discussed in relation to an aberrant neuronal sprouting that occurs around the degenerating motoneurons in the ventral horn during the course of the motoneuron disease.

Changes in receptor levels for thyrotropin releasing hormone, serotonin, and substance P in cervical spinal cord of Wobbler mouse: a quantitative autoradiography study during early and late stages of the motoneuron disease

Receptor levels for thyrotropin releasing hormone (TRH) measured by quantitative autoradiography in the Wobbler mouse cervical spinal cord show receptor losses that may relate to the inherited loss of motoneurons, most pronounced late (at Stage 4) in the motoneuron disease. An age-related decrease of TRH and serotonin (5-HT) receptors can be seen in the ventral horn of the control specimens (normal phenotype littermate and wild-type alike). However, this pattern is missing for substance P (SP) receptors from the wild-type specimens. Therefore the age-related decrease of SP receptors detected in the Wobbler mouse strain may identify a strain-related defect in SP neuronal/receptor developmental patterns. A higher level of TRH receptors was measured in the Wobbler dorsal horn at an early stage (Stage 1) in the motoneuron disease compared with the control specimens. The data are discussed in relation to an aberrant neuronal sprouting that occurs around the degenerating motoneurons in the ventral horn during the course of the motoneuron disease.

In situ hybridization analysis of myelin gene transcripts in developing mouse spinal cord

We analyzed the location and abundance of transcripts for the 4 CNS myelin protein genes, myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG), and 2',3'-cyclic nucleotide phosphohydrolase (CNP), in the mouse cervical spinal cord from the time of rapid myelination until adulthood (8-45 d). In the white matter, maximal levels of transcripts were found for each of the myelin genes at the peak of myelination (8 d). Total MBP and PLP mRNAs stayed high until 20 d and showed a minor decrease thereafter. In contrast, MAG and the MBP exon 2 containing transcripts (coding for the 21.5 and 17 kDa MBP isoforms) decreased sharply between 8 and 20 d, suggesting that high levels of these transcripts are needed primarily during the initiation of myelination. CNP transcripts were less abundant, maintained high expression until 20 d, and then decreased sharply. PLP, MAG, and CNP transcripts were clustered in the oligodendrocyte cell body, while MBP mRNAs were scattered throughout the cell body and processes. In contrast to the white matter, all these myelin specific transcripts in the gray matter showed a marked increase from 8 to 20 d, as did the number of oligodendrocytes identified by CNP immunostaining. MAG transcripts were found in white matter and in satellite and other oligodendrocytes of the gray matter but not in neurons identified by their expression of neurofilament transcripts. The results of our quantitative in situ hybridization study are in good agreement with those of previous molecular studies and provide new information on the cellular and topographic distribution of myelin-specific mRNAs during myelination.

Axonal growth cones and developing axonal collaterals form synaptic junctions in embryonic mouse spinal cord.

Axonal growth cones and developing axonal collaterals have been studied in electron microscopic and Golgi preparations of embryonic mouse cervical spinal cord. These structures are first observed on embryonic day 11 (E11), and by E12 both axonal growth cones and developing collaterals are observed to be the presynaptic elements of developing synaptic junctions. These relatively undifferentiated portions of axons comprise nearly one-half of the presynaptic components of the synaptic contacts observed on E12, and they continue to constitute about this same proportion during the rest of the embryonic period examined (E13–16). These early synaptic interactions of developing axonal collaterals may be involved in establishing the basic ramification patterns of mature axons by determining which developing collaterals will continue to grow and differentiate. Coated vesicles are observed fused with axonal collateral plasma membranes at the perimeters of developing synaptic contacts. Occasionally thin extensions of postsynaptic surfaces protrude into these coated vesicles, and similar relationships are also observed between presynaptic surfaces and coated vesicles fused with the membranes of postsynaptic elements. Such coated vesicle/ synaptic surface relationships may represent an early stage in the endocytosis of synaptic membranes by both the pre- and postsynaptic elements of developing synaptic contacts. This might represent a mechanism for the exchange of ‘molecular information’ between the components of protosynaptic contacts, and the results of such an exchange could be involved in determining whether protosynaptic contacts will differentiate into synaptic junctions. Developing axonal collaterals exhibit a preferential growth toward the sources of potential postsynaptic elements, and this growth implies that the axons present in the embryonic marginal zone are actively searching for appropriate postsynaptic processes rather than passively awaiting their arrival. Many of the collaterals weave around numerous intervening processes in order to reach their postsynaptic destinations, and unusual subaxolemmal lamellae are commonly observed where collaterals make acute curvatures around neurites which block their direct access to postsynaptic processes. The location of these lamellae suggests that they could be involved in producing changes in the direction of growth as developing axonal collaterals search for synaptic partnerships. Main axonal shafts commonly exhibit synaptic contacts adjacent to where they give rise to developing collaterals, and this observation hints that the formation of synapses by axonal shafts might stimulate collateral sprouting. However, other observations (see text) indicate that the formation of synaptic contacts by axonal shafts is not an obligatory prelude to collateral development. Therefore synaptic interactions of main axonal shafts do not appear to be a requirement for the primary induction of collateral development, but they may facilitate collateral growth by providing a confirmation that axons are located in appropriate synaptogenic fields.

The fine structure of the cervical spinal cord, ventral root and brachial nerves in the wobbler (wr) mouse.

Much indirect evidence supports the prevailing concept that the wobbler mouse represents an inherited form of lower motor neuron degeneration, with resultant neurogenic muscular atrophy, due to an autosomal recessive gene (wr). This report, which emphasizes the fine structural changes in the wobbler mouse cervical spinal cord, ventral root and brachial nerves, both extends and modifies previously published data concerning the ultrastructural changes in this disorder. In addition to the more obvious neuronal cytoplasmic vacuolization known to occur, studies extended to a broader age range of wobbler mice have revealed neuronal alterations such as nonvacuolar dissolution of granular endoplasmic reticulum, proliferation of cytoplasmic microtubules and neurofilaments, proliferation of branched tubules resembling smooth endoplasmic reticulum, development of lipid droplets, and increased numbers of pleomorphic dense and lamellar cytoplasmic bodies. The spectrum of pathological changes in this murine form of motor neuron disease thus appears more complex and varied than has hitherto been appreciated, and may be relevant to the formulation of pathogenetic hypotheses.

The mammalian red nucleus.

The mammalian red nucleus.