Exercise effects on the size and metabolic properties of soleus fibers in hindlimb-suspended rats.

The soleus atrophies rapidly when a rat is subjected to hindlimb suspension (HS), probably as a result of a decrease in the force encountered by the muscle. To test this premise, adult female rats were HS and half the rats were exercised (HS-EX) on a treadmill for 1.5 h.d-1 at 20 m.min-1 and a 30% grade. After 4 weeks, the midbelly of the soleus was prepared for histochemical analysis. Fibers were typed as dark or light staining for myosin ATPase, alkaline preincubation. Fiber size and quantitative histochemical enzyme activities of succinate dehydrogenase (SDH) were determined using a computer enhanced image processing system. In comparison to age-matched controls, the soleus wet weight was 69 and 30% smaller in HS and HS-EX rats. The mean cross sectional area of the dark ATPase fibers was reduced by 46 and 18% and light ATPase fibers by 69 and 48% in the HS and HS-EX, respectively. The percent dark ATPase fibers increased from 10% in the control rats to 19 and 17% in the HS and HS-EX. In both suspended groups, SDH activities in light ATPase fibers were 40% higher than control. The SDH activity of the dark ATPase fibers of HS-EX was 20% higher than control, while the dark ATPase fibers of HS were similar to control. To determine the degree to which these increases in SDH could be related to reductions in fiber size rather than increases in the actual amount of protein, integrated activity (activity/min x area) was calculated per fiber.(ABSTRACT TRUNCATED AT 250 WORDS)

Fiber type and fiber size of cat ankle, knee, and hip extensors and flexors following low thoracic spinal cord transection at an early age.

Effects of low thoracic spinal transection on muscle weights, fiber type distributions, and fiber cross-sectional areas (CSAs) of selected cat hind limb mixed-fast flexors and extensors and slow extensors were studied. Cats were spinalized at T12 at 2 weeks of age and maintained for 6 to 12 months. In general spinalization resulted in a decrease in muscle weights of extensors, while the weights of those muscles that function as either flexors or as both flexors and extensors were maintained. The percentages of fast-twitch [fast glycolytic (FG) and fast oxidative-glycolytic (FOG)] fibers increased and slow oxidative (SO) fibers decreased as a result of spinalization. However, FG fibers had a smaller CSA after spinal transection in both extensors and flexors. The total relative CSA of FG fibers per whole muscle was similar in spinalized and control cats. The relative muscle CSA occupied by SO fibers decreased in extensors and flexors as a result of a lower proportion of SO fibers and/or smaller SO fibers in spinalized cats. These findings suggest that muscles become more "fast-like" histochemically while little change occurs in the oxidative staining properties in either extensors or flexors in 6- to 12-month-old cats transected at 2 weeks of age. Further, these data suggest that the amount of muscle atrophy that occurs as a result of spinal transection is not closely related to the percentage reduction in SO fibers.

A computer model of the interleukin-4/receptor complex.

Interleukin-4 is a member of the cytokine family, a group of related messenger proteins which collectively help to moderate and control the immune response. It is believed that the folding topology of the beta-sheets of the interleukin-4 receptor (IL4R) is the same as that seen in the crystal structure of CD4. Although the sequence identity is low, homology modeling techniques have been used to model the IL4R structure from CD4. Refinement by molecular dynamics leads to a suggested structure which has been docked to interleukin-4 (IL4). Several residues of apparent importance for binding are identified.

Mutation detection in exons 1-14 of the adenomatous polyposis coli gene: identification of an alternatively spliced transcript.

Mutations in the APC gene are responsible for the dominantly inherited colon cancer syndrome, familial adenomatous polyposis (FAP). We have designed PCR primers which allow amplification by RT-PCR of exons 1-14 of the APC gene in six overlapping segments. The amplicons have been screened for the presence of mutations in patients affected with FAP using heteroduplex analysis. One patient has been identified with an alternatively spliced transcript involving exon 14 and a single base insertion mutation within the same exon.

Multiple interacting gene products may influence susceptibility to malignant hyperthermia.

Malignant hyperthermia (MH) is a potentially lethal disorder triggered in susceptible individuals on exposure to common anaesthetic agents. Crises reflect the consequences of disturbed skeletal muscle calcium homeostasis. MH is an autosomal dominant, genetically heterogeneous trait. Defects in a single major gene have been assumed to determine susceptibility status in individual families. However, in some pedigrees phenotypic and genotypic data are discordant. One explanation, in contrast to the current genetic model, is that susceptibility is dependent upon the effects of more than one gene. Using the transmission disequilibrium test we assessed the involvement of 8 MH candidate loci (RYR1, CACNA1S, CACNA2D1, MHS4 at 3q13.1, MHS6 at 5p, LIPE, DM1, dystrophin) by analysis of data from 130 MH nuclear families. Results suggested that variations in more than one gene may influence MH susceptibility in single families.

The G1021A substitution in the RYR1 gene does not cosegregate with malignant hyperthermia susceptibility in a British pedigree.

A single base change in the RYR1 gene encoding the skeletal muscle ryanodine receptor (calcium-sensitive calcium-release channel of the sarcoplasmic reticulum), resulting in the substitution of G1021 by A, has been proposed to underlie malignant-hyperthermia (MH) susceptibility in as many as 10% of cases in the European population. As part of our mutation-screening program in MH-susceptible (MHS) individuals, we have investigated this substitution in individuals from 151 unrelated British MHS families and have detected G1021A heterozygotes in 7 families. This mutation was not found in 156 unrelated MH-negative (MHN) individuals. We also examined eight families with central core disease (CCD): the mutation did not occur in any family members of any disease status (affected or unaffected for CCD, MHS, or MHN). In one large family, the G1021A mutation was found but did not show complete cosegregation with MH susceptibility: it occurred in only 7/12 MHS individuals in the kinship, and susceptibility was inherited from parents who were G1021 homozygotes, as well as from parents who were heterozygotes. On the basis of these findings, it is clearly unreliable at present to offer presymptomatic DNA testing for MH status, even in families in which a mutation has been detected.

Genetic heterogeneity and HOMOG analysis in British malignant hyperthermia families.

Malignant hyperthermia (MH) is an autosomal dominant genetic condition that presents in susceptible people undergoing general anaesthesia. The clinical disorder is a major cause of anaesthetic morbidity and mortality. The UK Malignant Hyperthermia Group has performed genetic linkage analysis on 20 large, well defined malignant hyperthermia families, using hypervariable markers on chromosome 19q13.1, including the candidate MH gene RYR1, the gene coding for the skeletal muscle ryanodine receptor protein. The results were analysed using LINKAGE to perform two point and multipoint lod scores, then HOMOG to calculate levels of heterogeneity. The results clearly showed genetic heterogeneity between MH families; nine of the families gave results entirely consistent with linkage to the region around RYR1 while the same region was clearly excluded in three families. In the remaining eight MHS families there were single recombinant events between RYR1 and MH susceptibility. HOMOG analysis was of little added benefit in determining the likelihood of linkage to RYR1 in these families. This confirmation of the presence of heterogeneity in the UK MH population, along with the possibility of the presence of two MH genes in some pedigrees, indicates that it would be premature and potentially dangerous to offer diagnosis of MH by DNA based methods at this time.

Producing exposed coat-free embryos.

Production of embryos that are free of tough outer coats facilitates studies that are not possible with embryos surrounded by impenetrable envelopes. This report describes a new procedure for preventing formation of fertilisation membranes in the sea urchin (Lytechinus pictus) model. This procedure involves treating unfertilised eggs with the enzyme alpha-amylase, which cleaves alpha-1,4 glucosidic bonds in the vitelline layer. A major advantage of this method is that it is very well defined and completely controllable with alpha-amylase inhibitor. The results suggest that intact alpha-1,4 glucosidic bonds are essential for vitelline layer integrity required for formation of the fertilisation membrane. Eggs treated with alpha-amylase possessed the same surface lectin receptors as untreated eggs and, as shown by light and transmission electron microscopy, produced healthy, cleaving embryos that were free of fertilisation envelopes.

Segregation of malignant hyperthermia, central core disease and chromosome 19 markers.

Malignant hyperthermia (MH) is an autosomal dominant disorder presenting under general anaesthesia. It is occasionally associated with a myopathy, central core disease (CCD), named after its predominant histochemical characteristic. The penetration of CCD is variable, but typically affected individuals show delayed motor milestones in infancy and remain physically compromised. It was thought until recently that individuals with CCD were always susceptible to MH. Individuals from eight CCD families were screened for the presence of 13 mutations in the skeletal muscle ryanodine receptor gene, reported previously to be associated with MH and/or CCD: none was detected. In seven of these families, where CCD and MH co-existed, we examined the segregation of CCD, MH susceptibility and chromosome 19q markers. In four families, there was complete co-segregation between MH, CCD and the chromosome 19 markers, but in one large pedigree there was a clear lack of segregation of CCD with either MH or chromosome 19 markers and there was no segregation between MH and these markers. This is unequivocal evidence that CCD, in common with MH, is genetically heterogeneous. In the two other families, CCD segregated with chromosome 19 markers but not all individuals with CCD were susceptible to MH. We recommend determination of MH susceptibility in all patients with CCD, irrespective of the MH status of their relatives with CCD.