Biomedical subjects

K L Knudsen

Publications and source records attributed to K L Knudsen.

17 recordsLinked to original sources

PubMed2004

Genetic basis of variation in morphological and life-history traits of a wild population of pink salmon.

Understanding the genetic basis of phenotypic variation is essential for predicting the direction and rate of phenotypic evolution. We estimated heritabilities and genetic correlations of morphological (fork length, pectoral and pelvic fin ray counts, and gill arch raker counts) and life-history (egg number and individual egg weight) traits of pink salmon (Oncorhynchus gorbuscha) from Likes Creek, Alaska, in order to characterize the genetic basis of phenotypic variation in this species. Families were created from wild-caught adults, raised to the fry stage in the lab, released into the wild, and caught as returning adults and assigned to families using microsatellite loci and a growth hormone locus. Morphological traits were all moderately to highly heritable, but egg number and egg weight were not heritable, suggesting that past selection has eliminated additive genetic variation in egg number and egg weight or that there is high environmental variance in these traits. Genetic correlations were similar for nonadjacent morphological traits and adjacent traits. Genetic correlations predicted phenotypic correlations fairly accurately, but some pairs of traits with low genetic correlations had high phenotypic correlations, and vice versa, emphasizing the need to use caution when using phenotypic correlations as indices of genetic correlations. This is one of only a handful of studies to estimate heritabilities and genetic correlations for a wild population.

Animals↗

PubMed2004

Linkage disequilibrium between the pseudoautosomal PEPB-1 locus and the sex-determining region of chinook salmon.

Allele frequency differences between sexes and an excess of heterozygotes in males had suggested that the PEPB-1 locus is sex-linked in chinook salmon (Oncorhynchus tshawytscha). We here estimate less than 1% recombination between PEPB-1 and a growth hormone pseudogene known to be in the sex-determining region (SEX) in 374 progeny from eight experimental matings. We present modified maximum likelihood methods for estimating haplotype frequencies from population samples at a sex-linked locus in which functional alleles occur on both the X and Y chromosomes (pseudoautosomal loci). We find nearly complete linkage disequilibrium between PEPB-1 and SEX in 20 population samples from the Puget Sound region of Washington and southern British Columbia. However, allele frequencies at PEPB-1 were similar in males and females in 35 population samples from the coast of Washington and the Columbia River basin. Pseudoautosomal regions have been described in a broad taxonomic array of vertebrates and invertebrates, and they are likely candidate regions to find genes associated with differences in life history, morphology, or behavior between males and females.

Animals↗

PubMed2000

Gene-centromere mapping of 312 loci in pink salmon by half-tetrad analysis.

We estimated recombination rates between 312 loci and their centromeres in gynogenetic diploid pink salmon (Oncorhynchus gorbuscha) that we produced by initiating development with irradiated sperm and blocking the maternal second meiotic division. Amplified fragment length polymorphisms (AFLPs) were significantly more centromeric than loci identified by three other techniques (allozymes, microsatellites, and PCR using primer sequences from interspersed nuclear elements). The near absence of AFLPs in distal regions could limit their utility in constructing linkage maps. A large proportion of loci had frequency of second division segregation (y) values approaching 1.0, indicating near complete crossover interference on many chromosome arms. As predicted from models of chromosomal evolution in salmonids based upon results with allozyme loci, all duplicated microsatellite loci that shared alleles (isoloci) had y values of nearly 1.0.

Animals↗

PubMed1994

One of two growth hormone genes in coho salmon is sex-linked.

Salmonid fishes have two growth hormone genes resulting from their polyploid ancestry. We used the polymerase chain reaction to examine genetic variation in the third intron (C) of both of these genes in coho salmon (Oncorhynchus kisutch). A polymorphism in the length of intron C in GH-1 is due to a variable number of copies of a 31-nt repeat that is absent from GH-1 of the closely related chinook salmon (Oncorhynchus tshawytscha) and rainbow trout (Oncorhynchus mykiss). Thus, this tandem repeat sequence has become established in the genome of coho salmon since the separation of this species from its closest relatives. All male coho salmon examined have an allele at the second growth hormone gene, GH-2, that is not found in females. GH-2 is thus on the sex chromosome and there is no recombination between GH-2 and the sex-determining locus (SEX). Sequences of intron C indicate much greater divergence between the X chromosome-specific allele and the Y chromosome-specific allele within coho salmon than between the X chromosome-specific alleles of coho and the closely related chinook salmon. Thus, absence of recombination between GH-2 and SEX apparently predates separation of these two species.

Alleles↗

PubMed1992

Sulfone-aromatic ligands for thiophilic adsorption chromatography: purification of human and mouse immunoglobulins.

New thiophilic matrices and new procedures were used for the purification of immunoglobulins both from human serum and from hybridoma cell cultures containing fetal calf serum. A range of aromatic and heteroaromatic ligands containing hydroxyl or amino groups have been coupled to divinyl sulfone-activated agarose. The resulting affinity matrices have the general formula M-O-CH2-CH2-SO2-CH2-CH2-X-Y, where M is the agarose matrix, X is oxygen or nitrogen, and Y is an aromatic or heteroaromatic compound. Contrary to earlier expectations these matrices showed pronounced thiophilic binding patterns when tested for the selective binding of immunoglobulins from human serum. The binding is influenced by the structure of the aromatic part of the ligand, the ligand concentration, and the concentration and type of lyotropic salt. 2-Hydroxypyridine coupled to divinyl sulfone-activated agarose was used to purify murine monoclonal antibodies (IgG1 and IgM) from hybridoma cell cultures containing fetal calf serum. Compared to previous methods, significantly increased binding capacity (300-1500%) was obtained by using 1.0-1.2 M ammonium sulfate. Purity of the monoclonal antibody may be optimized for each individual clone by washing the column with either a low concentration of ammonium sulfate or polyethylene glycol before elution.

Animals↗

PubMed1992

The autocrine role of tumor necrosis factor in the proliferation and functional differentiation of human lymphokine-activated T killer cells (T-LAK) in vitro.

The autocrine role of tumor necrosis factor alpha (TNF) in the proliferation and functional differentiation of human lymphokine-activated T-killer cells (T-LAK) in vitro was investigated. Human peripheral blood lymphocytes initially stimulated with IL-2 and phytohemagglutinin-P (PHA) for 48 h will proliferate for long periods in vitro in the presence of IL-2. These T-LAK cells have been shown to be 95% CD3 positive. Employing ELISA techniques, greater than 500 pg/ml of TNF was found to be released in the supernatants of these cells during the first 5 days of culture. However, the levels dropped to 100-200 pg/ml by days 7-10. T-LAK cells grown from days 7 to 10 in the presence of IL-2 and rabbit anti-TNF were significantly growth inhibited (up to 23%). The cytolytic activity of T-LAK cells grown from days 0 to 7 in the presence of anti-TNF was also decreased (up to 75%). Phenotypic analysis of these anti-TNF treated T-LAK cells revealed a decrease in CD8 expression (up to 12%) and increase in CD4 expression (up to 27%) when compared with control cells. The data suggest that TNF has a regulatory role in the growth and functional differentiation of these human T-LAK cells.

CD4 Antigens↗

PubMed1990

Temperature regulation and basal metabolic rate in the spotted skunk, Spilogale putorius.

1. Metabolic rate, body temperature, and evaporative water loss of six spotted skunks were measured at air temperatures between 8 and 40 degrees C. 2. The mean metabolic rate of spotted skunks at thermoneutral air temperatures was 30.5% below that predicted by body mass. 3. Thermal conductance, body temperature, and rates of evaporative water loss were like those of similar sized mammals. 4. The non-elongate body form, omnivorous diet, and low level of activity of spotted skunks distinguish them from other mustelids and may account for their lower-than-expected basal metabolism.

Animals↗

PubMed1988

Developmental rate and viability of rainbow trout with a null allele at a lactate dehydrogenase locus.

We show that a previously described isozyme polymorphism in rainbow trout (Salmo gairdneri) is the result of an enzymatically inactive (i.e., null) allele (n). Ldh3 null homozygotes (n/n) and heterozygotes (100/n) have reductions of about 20 and 12% in total lactate dehydrogenase (LDH) activity at hatching, respectively. As juveniles, (100/n) fish have reductions in LDH activity of 15, 37, and 21% in brain, heart, and white muscle, respectively. Embryos with different Ldh3 phenotypes from 11 families do not differ significantly in either survival or hatching time. However, a second measure of developmental rate, the amount of malate dehydrogenase (MDH) and phosphoglucomutase (PGM) activity in 33-day-old embryos, suggests that (100/n) embryos develop more slowly than (100/100) embryos. In three of four families examined, (100/n) embryos have significantly lower amounts of total MDH activity (8-10%). In one of these, (100/n) embryos also have significantly lower total PGM activity (15%). These data suggest that the reduction in total LDH activity is associated with small but detectable delays in developmental rate but nondetectable differences in survival to hatching.

Aging↗

PubMed1985

Heterozygosity and developmental stability in gynogenetic diploid and triploid rainbow trout.

We have previously reported that rainbow trout (Salmo gairdneri) more heterozygous at enzyme loci generally show increased developmental stability, as measured by reduced fluctuating asymmetry. We experimentally produced gynogenetic diploid and triploid individuals to test the effect of extreme heterozygosities on developmental stability. Gynogenetic diploids are identical by descent at an estimated 34 per cent of all loci and show a 50 per cent increase in the mean proportion of traits asymmetric per individual compared to normal rainbow trout. Triploids from two different strains have an estimated 30 per cent increase in genomic heterozygosity and a 14 per cent decrease in the mean proportion of traits asymmetric per individual compared to normal diploids. These changes in asymmetry induced by gynogenesis and triploidy are not as great as we expected based on the association between heterozygosity and fluctuating asymmetry in random mating populations. The narrow range of mean asymmetry in rainbow trout may result from an upper limit restricted by directional selection for developmental stability and a lower limit determined by inherent randomness in the developmental process.

Animals↗

PubMed1984

Major morphological effects of a regulatory gene: Pgm1-t in rainbow trout.

We have investigated the morphological effects of a genetic locus, Pgm1-t, that affects the expression of a phosphoglucomutase locus (Pgm1) in liver of rainbow trout (Salmo gairdneri). We have previously shown that embryos with liver Pgm1 expression hatch earlier than those without liver Pgm1 expression. We predicted that this difference in developmental rate should cause a reduction in meristic counts in the more rapidly developing fish with liver Pgm1 expression. Eight meristic (countable) characters in nine full-sib groups segregating for the presence or absence of liver Pgm1 expression are in agreement with this prediction. In eight of the nine families, there is a significant difference in the multivariate distribution of the eight meristic counts between full sibs with and without liver Pgm1 expression. This separation in multivariate space is based on a tendency for lower meristic counts in fish with liver Pgm1 expression. The magnitude of these morphological differences is similar to that between two subspecies of cutthroat trout (Salmo clarki) that show substantial genetic divergence at structural loci encoding enzymes (Nei's D = 0.34). These data support the view that small changes in the developmental process caused by genetic differences at regulatory genes can have large effects on morphology.

Animals↗

PubMed1983

Developmental stability and enzyme heterozygosity in rainbow trout.

The developmental pathways of organisms are genetically adjusted to produce the characteristic morphology of the species regardless of variations in internal and external conditions during development. This 'developmental buffering', however, is not always precise. Bilateral characters of an organism are often asymmetric, that is, different in size, shape or number. Fluctuating asymmetry occurs when the difference between a character on the left and right sides of individuals is normally distributed about a mean of zero. This type of asymmetry results from the inability of an organism to develop precisely along determined paths and can be used as a measure of developmental stability. Increased developmental stability would be reflected by reduced amounts of fluctuating asymmetry. We have now examined the relationship between the amount of fluctuating asymmetry for five bilateral characters and heterozygosity at 13 polymorphic loci in a population of rainbow trout (Salmo gairdneri). Our results indicate a significant negative correlation between the proportion of heterozygous loci and the proportion of asymmetric characters. These data provide evidence that individuals with greater heterozygosity within a population have increased developmental stability.

Animals↗

PubMed1983

Adaptive significance of differences in the tissue-specific expression of a phosphoglucomutase gene in rainbow trout.

We have investigated the phenotypic effects of a mutant allele that results in the expression of a phosphoglucomutase locus (Pgm1) in the liver of rainbow trout. Embryos with liver Pgm1 expression hatch earlier than embryos without liver Pgm1 expression. These differences apparently result from increased flux through glycolysis in embryos with liver PGM1 activity while they are dependent on the yolk for energy. Fish with liver PGM1 activity are also more developmentally buffered, as indicated by less fluctuating asymmetry of five bilateral meristic traits. The more rapidly developing individuals begin exogenous feeding earlier and achieve a size advantage that is maintained until sexual maturity. This size advantage produces a significant tendency for earlier age of sexual maturity. These results show that different genotypes at this regulatory gene result in important phenotypic differences that are likely to be of important adaptive significance.

Age Factors↗

PubMed1983

Structural and regulatory variation of phosphoglucomutase in rainbow trout.

The results of inheritance experiments with allozymic variants indicate that two pairs of duplicate loci encode PGM in rainbow trout. Two of the loci (Pgm-3,4) share five electromorphs and are highly polymorphic. The large number of phenotypes and instability of these isozymes make them difficult to score in population studies. The other pair of duplicate loci (Pgm1 and Pgm2) have diverged both structurally and in their patterns of tissue-specific expression. We have detected four electromorphs at Pgm2; this locus is expressed approximately equally in all tissues examined. Two electromorphs and a null allele have been detected at Pgm1. PGM1 activity is greatest in skeletal muscle, heart, and brain; only weak activity, if any, is detectable in liver, eye, stomach, and kidney. Ten percent of the trout from the Arlee strain have a greater than 100-fold increase in the expression of Pgm1 in the liver but have normal expression of this locus in other tissues. Results of genetic crosses are consistent with a single regulatory gene (Pgm1-t) with additive inheritance being responsible for the differences in liver PGM1 activity. The allele responsible for the expression PGM1 in the liver is rare in rainbow trout and is apparently a recent mutation. The presence of PGM1 liver activity has a variety of phenotypic effects that are likely to be of adaptive significance. Embryos with liver PGM1 activity develop more quickly than their full-sibs lacking activity. This difference apparently results from increased flux through glycolysis in embryos with liver PGM1 activity while they are dependent on the yolk for energy. The more rapidly developing individuals begin exogenous feeding earlier and obtain a size advantage that is maintained until sexual maturity. This size advantage also produces a tendency for earlier age of first sexual maturity. Fish with liver PGM1 activity are also more developmentally buffered, as indicated by less fluctuating asymmetry of five meristic traits.

Alleles↗

PubMed1982

Identification of a gene regulating the tissue expression of a phosphoglucomutase locus in rainbow trout.

Nine percent of the rainbow trout (Salmo gairdneri) from a hatchery source have a greater than 100-fold increase in expression of a phosphoglucomutase (PGM) locus, Pgm1, in the liver but have normal expression of this locus in other tissues. The results of genetic crosses are consistent with a single regulatory gene with additive inheritance being responsible for the differences in the amount of PGM activity in the liver.--The allele responsible for the expression of Pgm1 in the liver is apparently a recent mutation. This is supported by its restricted distribution in rainbow trout and the absence of liver Pgm1 expression in closely related species. This genetic system is valuable for future analysis of the control of gene expression and in determining the relative evolutionary importance of genetic variation at structural and regulatory genes.

Animals↗

PubMedDate not supplied

Specific interchain cross-linking of antibodies using bismaleimides. Repression of ligand leakage in immunoaffinity chromatography.

The extensive use of antibody-containing affinity columns in the purification of biologically active compounds (e.g., genetically engineered proteins) is severely hampered by the leaching of antibody (or portions thereof) from the immunoaffinity resin during elution of the target antigen. One of the major problems in this context is the combined use of reducing (i.e., thiols) and chaotropic (e.g., detergents and denaturants) agents in the elution step, which causes the disassociation of heavy and/or light chains from the immobilized antibody, thereby contaminating the resultant product. In order to overcome this problem, we have cross-linked the four antibody chains at their sites of disulfide interlinkage, thus producing a single antibody chain. To accomplish this, interchain disulfide bonds were reduced, and the resultant thiol groups were cross-linked by using bifunctional SH-specific reagents (particularly bismaleimides). Cross-linking of up to 95% of the available SH groups produced was achieved with concomitant retention of antigen-binding activity. The cross-linked antibody was immobilized onto CNBr-activated Sepharose, and the resultant column was found to be substantially more stable to harsh elution conditions than similar columns which contain the un-cross-linked antibody.

Antibodies↗

PubMedDate not supplied

Improved method for preparing N-hydroxysuccinimide ester-containing polymers for affinity chromatography.

N,N,N',N'-Tetramethyl(succinimido) uronium tetrafluoroborate is proposed as a reagent of choice for the activation of carboxyl groups and formation of N-hydroxysuccinimide esters on polymers. Unlike conventional methods which generate unstable gels, the reaction is appropriate for hydroxy-containing resins like Sepharose, cellulose, and dextran. The yields of activation and subsequent coupling capacity for ligands and proteins are very high. The respective columns can be used for affinity chromatography and immobilization of proteins.

Chromatography, Affinity↗

PubMedDate not supplied

Inheritance of nuclear DNA markers in gynogenetic haploid pink salmon.

We describe the inheritance of 460 PCR-based loci in the polyploid-derived pink salmon (Oncorhynchus gorbuscha) genome using gynogenetic haploid embryos. We detected a length polymorphism in a growth hormone gene (GH-2) intron that is caused by an 81 bp insertion homologous to the 3' end of the salmonid short interspersed repetitive element (SINE) SmaI. Such insertion polymorphisms within species bring into question the use of SINEs as phylogenetic markers. We confirmed that a microsatellite locus encodes a PCR-null allele that is responsible for an apparent deficit of heterozygotes in a population sample from Prince William Sound. Another set of microsatellite primers amplified alleles of the same molecular weight from both loci of a duplicated pair. In our analysis of several PCR-based multilocus techniques, we failed to detect evidence of comigrating fragments produced by duplicated loci. Segregation analysis of PCR-based markers using gynogenetic haploid embryos ensures that the interpretation of molecular variation is not complicated by heterozygosity, diploidy, or gene duplication. We urge investigators to test the inheritance of polymorphisms in salmonids prior to using them to measure genetic variation.

Animals↗