Merrill K. Wolf

Hypomyelinated mutant mice: Description of jpmsd and comparison with jp and qk on their present genetic backgrounds

Hypomyelinated mutant mice are valuable natural animal systems for analysis of CNS myelin development, chemistry and diseases. One mutant, jpmsd, has never received thorough morphological description. We here describe the detailed morphology of jpmsd, compare it with well-studied jp and qk on their present genetic backgrounds, and discuss the genetic histories of all 3 mutants. Region for region, jpmsd has twice as much myelin as jp, but 1/2--1/5 as much as qk. Both jp and jpmsd have scarce oligodendrocytes, rare nodes of Ranvier, clustering of myelin segments, abnormal lipid-filled cells, frequent degenerating cells, and rare distorted myelin profiles. In contrast, qk has abnormally numerous oligodendrocytes, frequent nodes of Ranvier, no obvious myelin clustering, no lipid-filled cells, rare degenerating cells, and frequent abnormal or distorted myelin profiles. jp and jpmsd are quantitatively different, but qualitatively similar. Since its origin, the jpmsd disease has inadvertently been ameliorated by transferring the mutation to a different background. Persistent differences in the remainder of the genome might account for all remaining apparent differences between jp and jpmsd. In contrast, qk is totally dissimilar in morphology and presumably in pathophysiology.

Shiverer jimpy double mutant mice. IV. Five combinations of allelic mutations produce three morphological phenotypes.

Mice which simultaneously express mutant genes at both the shi and jp loci (double mutant mice) have phenotypes much more complex than simple addition of individual mutant characteristics. Morphological study of shi jpmsd, shimld jp, and shi/shimld jp, and comparison with previously studied shi jp and shimld jpmsd, shows that 3 classes of central nervous system (CNS) white matter morphology are produced. (1) shi jp shows suppression of most jp characteristics: it is like shi except for more major dense line, and possibly more myelin, than shi alone. No other combination has as much myelin or any major dense line at all. (2) shi jpmsd has qualitative and quantitative characteristics intermediate between the two single mutants. (3) All combinations studied involving shimld have much less myelin than either single mutant. Qualitatively they express most jp locus features but suppress all shi locus features except abnormalities of myelin compaction. The difference between shi and shimld has more influence on the double mutant morphology than the difference between jp and jpmsd. In the 3-mutant combination, shi/shimld jp, the influence of shimld completely overrides that of shi. These morphological phenotypes resist assignment to any hierarchy of normalcy, and their specific features have no simple explanation in presently known molecular biology of the shi and jp locus mutations. They suggest the possibility of multiple copies and multiple primary functions of the messages at these loci.

Myelin deficient (shimld) mutant allele: Morphological comparison with shiverer (shi) allele on a B6C3 mouse stock

A new B6C3 stock of shimld mutant mice is compared in terms of behavior and CNS morphology with both a B6C3 shi stock and reports on other shimld animals. Defects of B6C3 shimld myelination seen at postnatal day 21 (P-21) are comparable to those in B6C3 shi with respect to % axons myelinated, sheath thickness, errors in the wrapping and targeting of myelin and abnormal oligodendrocyte shape. The two mutations are similarly expressed in cerebellar organotypic cultures. However, the major dense line (MDL) is present in a few shimld myelin sheaths at P-21 and a few sheaths show myelin basic protein by immunocytochemistry, while neither phenomenon is seen in shi at this age in the same CNS regions. Shimld mice survive their disease significantly better than shi. The shimld stock currently under study elsewhere differs from this B6C3 stock in that MDL was reported only in older animals, and behavior and survival were severely compromised.

Hypomyelinated mutant mice. V. Relationship between jp and jpmsd re-examined on identical genetic backgrounds

jp and jpmsd, two allelic mutations in the mouse that sharply reduce the amount of CNS myelin, produce diseases that can be distinguished morphologically only by their severity. This has raised the question of whether the two mutations are truly distinguishable. Since the two mutations have never been maintained on the same genetic background, correct quantitative and morphological comparison have not been possible. We have prepared a B6C3H stock of jp on the same genetic background as the available stock of jpmsd. In this jp stock, behavioral abnormalities, relative proportion of myelinated axons, and major morphological characteristics of the disease in situ are unchanged from the previous jp stock. The jp disease continues to be more severe than that of jpmsd. However, tissue from the new B6C3H stock myelinates better in organotypic culture than previous jp stocks. The increase in myelination is advantageous, not only for accurate comparison of the two alleles but for all culture studies of jp. Strictly comparable strains or stocks should be utilized in any comparative studies of closely related mutations such as jp and jpmsd.

Shiverer*jimpy double mutant mice. II. Morphological evidence supports reciprocal intergenic suppression

Mice which carry both the shiverer (shi) and the jimpy (jp) mutations have a morphological phenotype with features of each single mutation by itself but in milder form: the number of myelin sheaths is increased relative to jp, the thickness of sheaths and amount of major dense line is increased relative to shi, and the abnormal, lipid-filled cells characteristic of jp are not seen. However, the abnormal bundles of oligodendrocyte microprocesses and errors in the targeting of myelination which characterize shi are not altered by the presence of the jp mutation. This morphological evidence suggests partial reciprocal intergenic suppression in shiverer jimpy double mutant mice and therefore agrees with conclusions based on biochemical data presented by Kerner and Carson (Brain Research, 374 (1986) 45-53).

Jimpy-4J Mouse Has a Missense Mutation in Exon 2 of the Plp Gene

We previously showed that the jimpy-4J mouse mutation is located on the X chromosome, in or closely linked to the proteolipid protein (Plp) gene. The phenotype is characterized by the most severe hypomyelination of any of the naturally occurring myelin mutant mice, sharp reduction in oligodendrocyte number, and virtual absence of PLP protein. Affected animals show tremor, seizures, and die at about 24 postnatal days. We now report that sequencing of Plp genomic and cDNAs identifies a single nucleotide substitution in exon 2 that predicts an Ala38Ser substitutions in a hydrophilic region of PLP/DM20 protein close to a transmembrane domain. This mutation occurs in a very different region of the mouse Plp gene than that jimpy-msd mutations, yet all three produce qualitatively similar phenotypes.

CNS hypomyelinated mutant mice (jimpy, shiverer, quaking): in vitro evidence for primary oligodendrocyte defects.

Myelin is the protein and lipid sheath, consisting of consecutive, spirally wrapped, compacted segments of cell membrane, which surrounds each large axon in vertebrates and causes saltatory conduction of the action potentials. Nerve fiber sheaths with certain features of myelin are found in other animals, notably in some arthropods. True myelin, however, seems to be confined to true vertebrates, from the elasmobranchs up, and is strongly conserved in vertebrate evolution. Myelin has unique ultrastructural and biochemical components; it is formed by unique cells (in the CNS of warm-blooded vertebrates, by the oligodendrocytes); and it has a unique relationship to another cell, for normally it only forms around part of a neuron, almost always the axon. All this implies the existence of numerous DNA sequences which encode, not only for the structural components of adult myelin, but also for various steps of its development. Because myelin is strongly conserved, information about this DNA obtained from any mammal is likely to be directly applicable to other mammals including man.

CNS Hypomyelinated Mutant Mice: Morphological and Tissue Culture Studies

Publisher Summary This chapter discusses the morphological and tissue culture studies of central nervous system (CNS) hypomyelinated mutant mice. The neurological mutations of the mouse represent experiments of nature that provide powerful assistance in the studies of brain development. Among these mutations, a special opportunity is offered by the group with the hypomyelination of the central nervous system (CNS). The genetically normal mouse is born with almost no CNS myelin. The clinical syndrome produced by these mutations is not evident until the second postnatal week or even later and is similar in every case. All the hypomyelinated affected animals exhibit a bobbing or bouncing tremor, approximately 3 beats/s, most marked in the hindquarters. Most of the affected animals begin to exhibit tremor between 10 and 14 days of age, when their littermates are developing stable stance and gait. A few days after the onset of tremor, affected animals start having tonic convulsions of 20–30 s duration. Sometimes animals become paraplegic after convulsions, apparently as a result of dorsal midline spinal cord hemorrhage (the mouse corticospinal tract is in the dorsal columns). Eventually, they die during a convulsion.

Rumpshaker Behaves like Juvenile-Lethal Plp Mutations When Combined with Shiverer in Double Mutant Mice

The phenotypes of double mutant mice whose genomes are homozygous for an Mbp (myelin basic protein) mutation and hemizygous for a juvenile-lethal Plp (proteolipid protein) mutation were compared in earlier studies. The results suggested that the shiverer Mpb mutation might have some unexplained ability to partially rescue oligodendrocytes (OLs) from the ‘death sentence’ that is imposed by the Plp mutations. Conversely, they also indicated that the juvenile-lethal Plp mutations may normalize shiverer OL morphology by reducing the numbers of microprocesses. The Plp mutation rumpshaker produces a mild hypomyelination without reduction in OL numbers and a normal lifespan. This report describes double mutant mice combining two Mbp mutations with rumpshaker, utilizing a common B6C3F1 hybrid-based genetic background. Initial studies on B6C3F1 rumpshaker optic nerve and spinal cord white matter showed unanticipated signs of OL death, with morphologic criteria suggestive of an apoptotic mechanism. In shiverer*rumpshaker double mutant mice, this small class of dying cells could not be identified. White matter morphology was similar to that of mice expressing only the shiverer mutation, except that OL microprocesses were far less abundant. This evidence suggests that, despite their distinctive phenotypic differences, rumpshaker may share more characteristics with the juvenile-lethal Plp mutations than has previously been recognized.

Shiverer∗jimpy double mutant mice. III. Comparison of shimld∗jpmsd and shi∗jp phenotypes demonstrates dissimilar interactions of allelic mutations

Double mutant mice, which are of the genotype shimld/shimld*jpmsd/Y and therefore express both the shimld and jpmsd mutations, have a CNS myelin protein composition which resembles shimld/shimld alone but not jpmsd/Y alone. The double mutant CNS white matter morphology shows much less myelin and major dense line than either shimld/shimld or jpmsd/Y, but has other features which resemble jpmsd/Y but not shimld/shimld. In contrast, the parallel double mutant shi/shi*jp/Y, which expresses the alleles shi and jp rather than shimld and jpmsd, has already been shown to have biochemical and morphological phenotypes which are consistent with each other, both being intermediate between shi/shi and jp/Y and therefore suggesting partial reciprocal intergenic suppression (Brain Research, 374 (1986) 45-53 and 54-62). To assist in explaining the apparent inconsistencies between the biochemical and morphological phenotypes of the shimld/shimld*jpmsd/Y double mutant and between interactions of allelic mutations at the shi and jp loci, a hypothesis of multiple primary gene functions at these two loci is proposed.