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Biomedical subjects

M Carlson

Publications and source records attributed to M Carlson.

At least 217 records · Page 12Linked to original sources

Effects of monocular exposure to oriented lines on monkey striate cortex.

This study examines the extent to which the restriction of visual experience to lines of a single orientation influences the organization of the striate cortex in infant monkeys (Macaca mulatta). Previous studies of kittens raised with monocular exposure to a single line orientation have consistently shown the response preference of cells driven by that eye to be biased towards the experienced orientation. Studies of binocular exposure to restricted orientations have been equivocal. In the infant monkey cortex responses to oriented lines have virtually all the specificity of responses seen in the adult animal. In an effort to clarify the phenomenon and the mechanism by which orientation bias might be obtained, we examined the effects of monocular exposure to a restricted orientation in infant macaques. Three monkeys were used. Each monkey was raised with one open eye exposed to lines of a single orientation and one eye occluded by lid suture. As in other cases of monocular deprivation in either cat or monkey, few binocularly driven cells were recorded and the majority of cells were dominated by the open eye. Cells driven by the open eye had normal representation of all orientation preferences and there was no overall increase in the number of cells preferring the orientation to which the eye had been exposed. The cells dominated by the occluded eye, however, showed a lack of cells responding to orientations to which the open eye had been exposed. These findings suggest that a competitive mechanism operates between the two eyes to provide an orientation selective advantage to the open eye.

Animals↗

Suppressors of SNF2 mutations restore invertase derepression and cause temperature-sensitive lethality in yeast.

Mutations in the SNF2 gene of Saccharomyces cerevisiae prevent derepression of the SUC2 (invertase) gene, and other glucose-repressible genes, in response to glucose deprivation. We have isolated 25 partial phenotypic revertants of a snf2 mutant that are able to derepress secreted invertase. These revertants all carried suppressor mutations at a single locus, designated SSN20 (suppressor of snf2). Alleles with dominant, partially dominant and recessive suppressor phenotypes were recovered, but all were only partial suppressors of snf2, reversing the defect in invertase synthesis but not other defects. All alleles also caused recessive, temperature-sensitive lethality and a recessive defect in galactose utilization, regardless of the SNF2 genotype. No significant effect on SUC2 expression was detected in a wild-type (SNF2) genetic background. The ssn20 mutations also suppressed the defects in invertase derepression caused by snf5 and snf6 mutations, and selection for invertase-producing revertants of snf5 mutants yielded only additional ssn20 alleles. These findings suggest that the roles of the SNF2, SNF5 and SNF6 genes in regulation of SUC2 are functionally related and that SSN20 plays a role in expression of a variety of yeast genes.

Enzyme Repression↗

Null mutations in the SNF3 gene of Saccharomyces cerevisiae cause a different phenotype than do previously isolated missense mutations.

Missense mutations in the SNF3 gene of Saccharomyces cerevisiae were previously found to cause defects in both glucose repression and derepression of the SUC2 (invertase) gene. In addition, the growth properties of snf3 mutants suggested that they were defective in uptake of glucose and fructose. We have cloned the SNF3 gene by complementation and demonstrated linkage of the cloned DNA to the chromosomal SNF3 locus. The gene encodes a 3-kilobase poly(A)-containing RNA, which was fivefold more abundant in cells deprived of glucose. The SNF3 gene was disrupted at its chromosomal locus by several methods to create null mutations. Disruption resulted in growth phenotypes consistent with a defect in glucose uptake. Surprisingly, gene disruption did not cause aberrant regulation of SUC2 expression. We discuss possible mechanisms by which abnormal SNF3 gene products encoded by missense alleles could perturb regulatory functions.

Cloning, Molecular↗

Molecular analysis of SNF2 and SNF5, genes required for expression of glucose-repressible genes in Saccharomyces cerevisiae.

The SNF2 and SNF5 genes are required for derepression of SUC2 and other glucose-repressible genes of Saccharomyces cerevisiae in response to glucose deprivation. Previous genetic evidence suggested that SNF2 and SNF5 have functionally related roles. We cloned both genes by complementation and showed that the cloned DNA was tightly linked to the corresponding chromosomal locus. Both genes in multiple copy complemented only the cognate snf mutation. The SNF2 gene encodes a 5.7-kilobase RNA, and the SNF5 gene encodes a 3-kilobase RNA. Both RNAs contained poly(A) and were present in low abundance. Neither was regulated by glucose repression, and the level of SNF2 RNA was not dependent on SNF5 function or vice versa. Disruption of either gene at its chromosomal locus still allowed low-level derepression of secreted invertase activity, suggesting that these genes are required for high-level expression but are not directly involved in regulation. Further evidence was the finding that snf2 and snf5 mutants failed to derepress acid phosphatase, which is not regulated by glucose repression. The SNF2 and SNF5 functions were required for derepression of SUC2 mRNA.

Cloning, Molecular↗

Short repeated elements in the upstream regulatory region of the SUC2 gene of Saccharomyces cerevisiae.

Expression of secreted invertase from the SUC2 gene is regulated by carbon catabolite repression. Previously, an upstream regulatory region that is required for derepression of secreted invertase was identified and shown to confer glucose-repressible expression to the heterologous promoter of a LEU2-lacZ fusion. In this paper we show that tandem copies of a 32-base pair (bp) sequence from the upstream regulatory region activate expression of the same LEU2-lacZ fusion. The level of expression increased with the number of copies of the element, but was independent of their orientation; the expression from constructions containing four copies of the sequence was only twofold lower than that when the entire SUC2 upstream regulatory region was present. This activation was not significantly glucose repressible. The 32-bp sequence includes a 7-bp motif with the consensus sequence (A/C)(A/G)GAAAT that is repeated at five sites within the upstream regulatory region. Genetic evidence supporting the functional significance of this repeated motif was obtained by pseudoreversion of a SUC2 deletion mutant lacking part of the upstream region, including two copies of the 7-bp element. In three of five pseudorevertants, the mutations that restored high-level SUC2 expression altered one of the remaining copies of the 7-bp element.

Base Sequence↗

Fecal alpha 1-antitrypsin and hemoglobin excretion in healthy human milk-, formula-, or cow's milk-fed infants.

There is concern that whole cow's milk feedings may be associated with intestinal abnormalities in infants. We studied this issue by measuring random fecal samples for alpha 1-antitrypsin (FA1AT) and hemoglobin (FH) concentrations in 820 healthy infants up to 12 months of age. Subjects were fed either human milk, formula, or fresh whole cow's milk. Solid foods were given ad libitum. Fecal samples were also tested for occult blood with Hematest reagent tablets. None of the infants younger than 6 months of age were receiving fresh whole cow's milk. We found small but statistically significant differences in mean FA1AT between the three feeding groups (P less than .0001): human milk (n = 354) greater than formula (n = 320) greater than cow's milk (n = 146). The younger subjects fed either formula or human milk tended to have higher FA1AT concentrations than did the age-matched subjects who were not consuming solid foods (P less than or equal to .005). Daily FA1AT excretion, FA1AT concentration, and daily stool output were subsequently determined on a separate group of 40 infants 8 to 12 months of age to ascertain whether differences in total daily FA1AT excretion occur in children fed different types of milk. Total daily FA1AT excretion was similar in the three milk feeding groups. An inverse correlation between FA1AT concentration and daily stool output was also found (P less than .001). The overall rate of detectable FH in 792 stool smears was 2.1% and unrelated to type of milk feeding. Of 705 stool smears, 3.5% had positive Hematest reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Comparison of two yeast invertase genes: conservation of the upstream regulatory region.

The yeast genome contains a dispersed family of invertase structural genes (SUC1-SUC5, SUC7). Five of these genes are located very close to telomeres and are flanked by large regions of homologous sequence; recombination between telomeres could account for the dispersal of these SUC genes to different chromosomes. The SUC2 locus, in contrast, is not near a telomere and does not share large regions of flanking homology with the other loci. We examine here the relationship between SUC2 and one of the telomeric genes, SUC7. Sequence comparison revealed homology extending from about position -624 to +1791, which is close to the end of the mRNA. The 5' noncoding sequence includes two highly conserved regions: the region between -140 and +1, which contains the TATA box and presumably other promoter elements, and a second region extending from -508 to -400, which corresponds to the upstream regulatory region.

Amino Acid Sequence↗

Rearrangement of the genetic map of chromosome VII of Saccharomyces cerevisiae.

The genetic map of the right arm of chromosome VII of Saccharomyces cerevisiae includes markers on a distal segment for which meiotic linkage to the centromere-proximal marker cly8 has not previously been demonstrated. According to the currently accepted map, SUF4 is the most distal marker on the right arm. We have shown by tetrad analysis that SUF4 is linked to cly8 and ade6. The genetic distance between SUF4 and cly8 is 29 cM. These data indicate that the genetic map of the right arm of chromosome VII should be revised by inverting the orientation of the distal segment so that SUF4 is located near cly8, and SUC1 and MAL1 are the most distal markers. With this revision, all of the polymeric fermentation markers that have been mapped are located at the ends of chromosomes.

Chromosome Mapping↗

Upstream region of the SUC2 gene confers regulated expression to a heterologous gene in Saccharomyces cerevisiae.

The SUC2 gene produces two differently regulated mRNAs that encode two forms of invertase. The 1.9-kilobase mRNA encoding secreted invertase is regulated by glucose (carbon catabolite) repression, and the 1.8-kilobase mRNA encoding intracellular invertase is synthesized constitutively. Previous work has shown that the 5' noncoding region between -650 and -418 is required for derepression of secreted invertase in response to glucose deprivation. We show here that this upstream region can confer glucose-repressible expression to a heterologous gene, a LEU2-lacZ gene fusion, that is not normally regulated by glucose repression. This expression was found to respond appropriately to mutations in trans-acting genes that affect regulation of SUC2 expression. Mutations in the SNF1 through SNF6 loci reduced derepression of beta-galactosidase, and a mutation at the SSN6 locus caused constitutive expression. These findings indicate that the SUC2 upstream region mediates the regulatory effects of these genes and suggest that regulation occurs at the level of transcription. In addition, the upstream region was partially active in the inverted orientation.

Chromosome Inversion↗

Evolution of the dispersed SUC gene family of Saccharomyces by rearrangements of chromosome telomeres.

The SUC gene family of Saccharomyces contains six structural genes for invertase (SUC1 through SUC5 and SUC7) which are located on different chromosomes. Most yeast strains do not carry all six SUC genes and instead carry natural negative (suc0) alleles at some or all SUC loci. We determined the physical structures of SUC and suc0 loci. Except for SUC2, which is an unusual member of the family, all of the SUC genes are located very close to telomeres and are flanked by homologous sequences. On the centromere-proximal side of the gene, the conserved region contains X sequences, which are sequences found adjacent to telomeres (C. S. M. Chan and B.-K. Tye, Cell 33:563-573, 1983). On the other side of the gene, the homology includes about 4 kilobases of flanking sequence and then extends into a Y' element, which is an element often found distal to the X sequence at telomeres (Chan and Tye, Cell 33:563-573, 1983). Thus, these SUC genes and flanking sequences are embedded in telomere-adjacent sequences. Chromosomes carrying suc0 alleles (except suc20) lack SUC structural genes and portions of the conserved flanking sequences. The results indicate that the dispersal of SUC genes to different chromosomes occurred by rearrangements of chromosome telomeres.

Base Sequence↗

Myoglobin in rat hind limb muscles after denervation and during reinnervation.

Radioimmunoassay of myoglobin (Mb) was performed in rat hind limb muscles after surgical denervation and during reinnervation following cryolesion of the sciatic nerve. Muscles of the contralateral leg served as controls. After resection of the sciatic nerve, decreased Mb concentrations were noted on the fourth day in the tibialis anterior, peroneus longus, and extensor digitorum longus (EDL) muscles. Thereafter, the levels increased up to the last observation on day 32. The increases in Mb levels in the tibialis anterior and EDL muscles were considerably more pronounced (305% and 324%, respectively) than in the peroneus longus and soleus muscles (148% and 137%, respectively). After cryolesion of the sciatic nerve, the Mb concentrations in the tibialis anterior, peroneus longus, and EDL muscles increased, reaching maximal values on days 16-21. The levels then decreased and normal values were observed 2 months postoperatively. The normalization of the Mb levels during reinnervation corresponded fairly well in time with the clinical recovery and neurophysiological findings observed in a previous study.

Animals↗

Development of tactile discrimination capacity in Macaca mulatta. I. Normal infants.

Infant macaques between the ages of 7 and 25 weeks of age were trained on a series of manual tactile discrimination tasks. Tactile discrimination capacity, as measured by the most difficult level of size and texture discrimination tasks mastered, was the same for all ages of infants and did not differ from that of adults. Infants as young as 10 weeks of age were found to have a discrimination capacity similar to that of adult macaques, although an adult level of manual motor control had not been achieved by this early age. During the acquisition of size tasks, older animals made fewer errors than did younger animals, suggesting an improved efficiency in size discrimination capacity over the first 6 months of life. By contrast, the efficiency with which the younger animals mastered texture discrimination was superior to that of the older infants. The possible contributions of sensory experience or manual motor control to the maturation of sensory capacity were examined by applying 16 weeks of sensory restriction in one infant and a unilateral motor cortex lesion in another infant, respectively. Only transient impairment was found in either case suggesting that neither tactile experience nor motor control contribute significantly to the maturation of tactile discrimination capacity in infant macaques.

Aging↗

Development of tactile discrimination capacity in Macaca mulatta. II. Effects of partial removal of primary somatic sensory cortex (SmI) in infants and juveniles.

Four infant macaques between the ages of 2.7 and 6.7 weeks and 5 juvenile macaques between the ages of 78.6 and 108.3 weeks received partial, unilateral lesions of the hand projection in primary somatic sensory cortex (SmI) (either Brodmann's area 3 or areas 1 and 2 combined). Following these partial SmI lesions, both infants and juveniles showed some initial impairment on acquisition and slightly inferior performance on the size-ALL task. Both infants and juveniles were able to discriminate smaller size differences with the contralateral hand than partial SmI-lesioned adult macaques with similar SmI lesions in previous studies. After partial SmI lesions, infants and juveniles were markedly retarded in the acquisition of texture discriminations indicating that, as in adult macaques, these separate SmI areas contribute jointly to texture discrimination capacity. However, in contrast to adult animals, all infants and most juveniles were able to master all levels of texture tasks after 2-4 months of training. On the texture-ALL task, the performance of partial SmI-lesioned infants was slightly inferior to that of normal infants with the contralateral hand but significantly better than normal infants on the ipsilateral hand. Lesioned juveniles showed impairment on both contralateral and ipsilateral hands. Recovery of near-normal tactile function in both partial SmI-lesioned infants and juveniles is seen although mature levels of function are typical of normal infants over this same age range.

Aging↗

Development of tactile discrimination capacity in Macaca mulatta. III. Effects of total removal of primary somatic sensory cortex (SmI) in infants and juveniles.

Four infant macaques between the ages of 3.0 and 5.1 weeks and three juvenile macaques between the ages of 79.9 and 109.3 weeks received unilateral lesions of all cytoarchitectural fields (Brodmann's areas 3, 1 and 2) in the hand area of the postcentral gyrus. These total SmI-lesioned infants acquired the size and texture tasks within the same time period and with the same efficiency as normal infants. On some size- and texture-ALL tasks they actually performed significantly better than partial SmI-lesioned or normal infants. The normal acquisition and ALL performance of the infants with total SmI lesions contrasts with that of the juveniles with comparable lesions. On size acquisition and ALL tasks, with the contralateral hand, total SmI-lesioned juveniles were significantly inferior to normal and total SmI-lesioned infants and to partial SmI-lesioned juveniles. During texture acquisition with the contralateral hand, the total SmI-lesioned juveniles made significantly more errors than normal and total SmI-lesioned infants. The capacity to recover from partial SmI lesions extends into the second year of life and is a gradual process which may be mediated by remaining SmI subdivisions. By contrast, the capacity to recover from total SmI lesions is restricted to infants and is a rapid process which must depend on other areas within the damaged, or possibly the intact, hemisphere.

Aging↗

A suppressor of SNF1 mutations causes constitutive high-level invertase synthesis in yeast.

The SNF1 gene product of Saccharomyces cerevisiae is required to derepress expression of many glucose-repressible genes, including the SUC2 structural gene for invertase. Strains carrying a recessive snf1 mutation are unable to ferment sucrose. We have isolated 30 partial phenotypic revertants of a snf1 mutant that were able to ferment sucrose. Genetic characterization of these revertants showed that the suppressor mutations were all recessive and defined eight complementation groups, designated ssn1 through ssn8 (suppressor of snf1 ). The revertants were assayed for secreted invertase activity, and although activity was detected in members of each complementation group, only the ssn6 strains contained wild-type levels. Synthesis of secreted invertase in ssn6 strains was found to be constitutive, that is, insensitive to glucose repression; moreover, the ssn6 mutations also conferred constitutivity in a wild-type ( SNF1 ) genetic background and are, therefore, not merely suppressors of snf1 . Pleiotropic defects were observed in ssn6 mutants. Genetic analysis suggested that the ssn6 mutations are allelic to the cyc8 mutation isolated by R. J. Rothstein and F. Sherman, which causes increased production of iso-2-cytochrome c. The data suggest a regulatory function for SSN6 .

Carbohydrate Metabolism↗

Genes affecting the regulation of SUC2 gene expression by glucose repression in Saccharomyces cerevisiae.

Mutants of Saccharomyces cerevisiae with defects in sucrose or raffinose fermentation were isolated. In addition to mutations in the SUC2 structural gene for invertase, we recovered 18 recessive mutations that affected the regulation of invertase synthesis by glucose repression. These mutations included five new snf1 (sucrose nonfermenting) alleles and also defined five new complementation groups, designated snf2, snf3, snf4, snf5, and snf6. The snf2, snf4, and snf5 mutants produced little or no secreted invertase under derepressing conditions and were pleiotropically defective in galactose and glycerol utilization, which are both regulated by glucose repression. The snf6 mutant produced low levels of secreted invertase under derepressing conditions, and no pleiotropy was detected. The snf3 mutants derepressed secreted invertase to 10-35% the wild-type level but grew less well on sucrose than expected from their invertase activity; in addition, snf3 mutants synthesized some invertase under glucose-repressing conditions.--We examined the interactions between the different snf mutations and ssn6, a mutation causing constitutive (glucose-insensitive) high-level invertase synthesis that was previously isolated as a suppressor of snf1. The ssn6 mutation completely suppressed the defects in derepression of invertase conferred by snf1, snf3, snf4 and snf6, and each double mutant showed the constitutivity for invertase typical of ssn6 single mutants. In contrast, snf2 ssn6 and snf5 ssn6 strains produced only moderate levels of invertase under derepressing conditions and very low levels under repressing conditions. These findings suggest roles for the SNF1 through SNF6 and SSN6 genes in the regulation of SUC2 gene expression by glucose repression.

Enzyme Repression↗

Cloning and genetic mapping of SNF1, a gene required for expression of glucose-repressible genes in Saccharomyces cerevisiae.

A functional SNF1 gene product is required to derepress expression of many glucose-repressible genes in Saccharomyces cerevisiae. Strains carrying a snf1 mutation are unable to grow on sucrose, galactose, maltose, melibiose, or nonfermentable carbon sources; utilization of these carbon sources is regulated by glucose repression. The inability of snf1 mutants to utilize sucrose results from failure to derepress expression of the structural gene for invertase at the RNA level. We isolated recombinant plasmids carrying the SNF1 gene by complementation of the snf1 defect in S. cerevisiae. A 3.5-kilobase region is common to the DNA segments cloned in five different plasmids. Transformation of S. cerevisiae with an integrating vector carrying a segment of the cloned DNA resulted in integration of the plasmid at the SNF1 locus. This result indicates that the cloned DNA is homologous to sequences at the SNF1 locus. By mapping a plasmid marker linked to SNF1 in this transformant, we showed that the SNF1 gene is located on chromosome IV. We then mapped snf1 to a position 5.6 centimorgans distal to rna3 on the right arm; snf1 is not extremely closely linked to any previously mapped mutation.

Chromosome Mapping↗