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Biomedical subjects

U Kuhnlein

Publications and source records attributed to U Kuhnlein.

At least 19 recordsLinked to original sources
PubMed2006

Development of a real-time (Q) PCR assay to measure variation in expression of prolactin receptor mRNA in the hypothalamus and pituitary gland during late embryogenesis in turkeys and chickens.

Changes in levels of PRLR mRNA in the pituitary gland and hypothalamus of chickens and turkeys from embryonic day (ED) 15 and ED21 to 1 day post-hatch, respectively, were measured by real-time PCR. In both species, PRLR mRNA increased from low levels during the last week of ED to reach maxima at the peri-hatch period. Similarly, circulating levels of PRL also increased during this interval and were highly correlated with levels of the PRLR mRNA in both the pituitary gland and hypothalamus. This suggests that PRL was up-regulating its receptor. In support of this, stimulation of the turkey pituitary gland with VIP on ED24 resulted in a 4- and 3-fold increase in PRL and PRLR, respectively. Since VIP had no direct effect on the levels of PRLR transcript in the hypothalamus, it is likely that VIP is acting indirectly through increased PRL to up-regulate the number of receptors.

Animals↗
PubMed2004

Ornithine decarboxylase: haplotype structure and trait associations in White Leghorn chickens.

Sequence analysis of 4,230 bp of the 3' end of the ornithine decarboxylase gene of 20 chickens from a noninbred White Leghorn strain revealed a total of 62 polymorphisms. Of these 61% were transitions, 24% were transversions, and 16% were deletions or insertions. Despite the high number of polymorphisms, only 3 haplotypes were present among the 40 alleles analyzed. Based on the genetic distances between haplotypes they segregated early during domestication of the chicken. Ornithine decarboxylase is a pivotal enzyme in regulating the synthesis of polyamines, cations that are important regulators of cell division, differentiation, and apoptosis. Variants of ornithine decarboxylase are, therefore, expected to affect many different traits. Association analyses between genotypes and the major egg production traits in female chickens of the same strain revealed significant effects on the onset of sexual maturity, BW at sexual maturity, eggshell thickness (a measure of calcium deposition), and residual feed consumption (a measure of the metabolic rate). Further, comparisons of the genotypes indicated that the 3 haplotypes differ in their phenotypic properties. Our results show that variations in a gene that is ubiquitously expressed in all cells of an organism may nevertheless contribute to distinct phenotypic properties of the organism as a whole.

Animals↗
PubMed2003

Alleles of cytosolic phosphoenolpyruvate carboxykinase (PEPCK): trait association and interaction with mitochondrial PEPCK in a strain of White Leghorn chickens.

White Leghorn chickens from a nonselected closed population were typed for two RFLP located in the 3' end of the gene coding for cytosolic phosphoenol-pyruvate carboxykinase (PEPCK-C), a major control gene of gluconeogenesis. The two RFLP gave rise to three alleles (or haplotype classes), thus defining six genotypes. Feed efficiency (FE) and residual feed consumption (RFC) varied significantly among the genotypes and indicated that all three haplotypes differed from each other. FE is the ratio between feed consumption and egg mass produced, whereas RFC is the feed consumption after correcting for BW and egg production. There was significant interaction between PEPCK-C genotypes and mitochondrial PEPCK (PEPCK-M) genotypes defined by a single RFLP. The latter enzyme catalyzes the same reaction but is located in the matrix of the mitochondria and is encoded by a different nuclear gene. Interaction was evident from an analysis of the egg weight and egg specific gravity in the early phase of egg laying. It was such that the effect of the variation in one gene depended entirely on the genotype of the second gene. In addition, significant genotypic disequilibria were observed between two of the three alleles of PEPCK-C and between one of these alleles and the two RFLP alleles of PEPCK-M. This finding indicates variations of genes in the gluconeogenesis pathway may affect feed utilization and egg production traits, as well as reproductive fitness.

Alleles↗
PubMed2003

The dynamics of the genotype-phenotype association.

The integrity of an organism is maintained by networks of interacting genes. Such networks predict that genetic variants affect phenotypes in a nonadditive fashion. That is, the effect of an allelic variation in one gene is dependent on the variations in other genes. We summarize the analyses of a series of genes in a White Leghorn strain that support the existence of such gene networks: 1) genes are pleiotropic, 2) genes affect trait correlations, 3) genes affect trait distributions in a nonadditive fashion, 4) genes interact with each other, and 5) genes are at linkage disequilibrium, even when located on different chromosomes. The latter observation indicated that certain gene combinations lead to reduced reproductive fitness. Each candidate genes we analyzed segregated for multiple alleles that affected production traits. This finding was surprising, even for a strain with a large effective population size. The shapes of trait distributions appear to be a better descriptor of gene effects than measures of central tendency. Despite this complexity, it is feasible to conduct DNA-based selection, starting from any of several different genes that affect a trait. Gene networks may be altered in many different ways to improve a particular phenotype, but networks may differ in their effects on other traits.

Animal Husbandry↗
PubMed2002

Genetic variability of the cytosolic phosphoenolpyruvate carboxykinase gene in white leghorn chickens.

Phosphoenolpyruvate carboxykinase (PEPCK) is a key regulatory enzyme of gluconeogenesis. Genetic variations in this gene may therefore affect a wide variety of traits, including tumor growth that is heavily dependent on glucose metabolism. We have previously shown the gene coding for the mitochondrial form of PEPCK (PEPCK-M) segregates for markers that are collected with resistance to Marek's disease. In this communication we analyze the genetic variability of PEPCK-C, the gene which codes for the cytosolic form of PEPCK. A 3,792-bp segment of 5'-region of the PEPCK-C gene (position -1723 to 2069) was sequenced in four individuals from eight different strains of White Leghorn chickens (a total of 64 genomes). A total of 19 single nucleotide polymorphisms (SNP) were identified. Neither deletions nor insertions were present. The most frequent SNP were transitions (79%), and in most cases the ancestral allele coincided with CpG dinucleotides (10-fold excess after correcting for dinucleotide frequencies). A gene tree was constructed assuming maximal parsimony. It led to the delineation of 6 haplotypes (combination of alleles). Two of the SNP coincided with RFLP detectable by the restriction enzymes AciI and BstEII, respectively. Based on this analysis we can now identify individuals with the evolutionary most distant PEPCK-C haplotypes, establish strains of these haplotypes, and analyze trait associations and epistasis with other genes.

Animals↗
PubMed1999

Cytogenetic mapping of 31 functional genes on chicken chromosomes by direct R-banding FISH.

Using direct R-banding fluorescence in situ hybridization, we determined the location of 31 functional genes on chicken chromosomes. Replication R-banded chromosomes were obtained by synchronizing splenocyte cultures with excessive thymidine, followed by BrdU treatment. Thirty-one functional genes were directly localized to banded chicken chromosomes using genomic DNA and cDNA fragments as probes. The possibility of conserved linkage homology between chicken and human chromosomes was demonstrated for seven chicken chromosome regions (1p, 1q, 2q, 4p, 4q, and 5q).

Animals↗
PubMed1998

Copulatory behaviour and paternity in the American kestrel: the adaptive significance of frequent copulations.

The adaptive significance of repeated withinpair copulations is not well understood. We analysed the copulatory behaviour of 16 pairs of solitary-nesting American kestrels, Falco sparverius, in southern Quebec (Canada), and the achieved reproductive success (paternity) of 21 kestrel families determined by DNA fingerprinting, in terms of four hypotheses. (1) The paternity assurance hypothesis, which suggests that males copulate frequently to avoid being cuckolded, was rejected because there were few extrapair copulation attempts (<1% of all copulations observed), withinpair copulations were not timed during the fertile period and mate attendance did not increase as the fertile period approached. (2) The immediate material benefits hypothesis, which suggests that females trade copulations for food, was refuted because copulations most often occurred without food transfers, especially outside the fertile period. (3) The female mate guarding of males hypothesis, which suggests that females distract their mates from other mating opportunities by copulating frequently, was rejected because extrapair copulation attempts were infrequent, male and female solicitation frequencies were similar and females did not differ in the timing or frequency of solicitations. (4) The mate assessment hypothesis, which suggests that assessment of mate quality is mediated via copulation, most closely predicted the behaviour observed since withinpair copulations were frequent outside the fertile period and at pair formation, males and females solicited similar numbers of copulations and pairs did not differ significantly in solicitation or copulation frequency. In line with this hypothesis we found that only one brood was extrapair, probably the result of mate replacement.Copyright 1998 The Association for the Study of Animal Behaviour

Journal Article↗
PubMed1998

Mitochondrial PEPCK: a highly polymorphic gene with alleles co-selected with Marek's disease resistance in chickens.

The gene coding for the mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M), a pivotal component in gluconeogenesis from lactate via the Cori cycle, was highly polymorphic in strains of egg-type chickens (White Leghorn) of different origins. Based on MspI restriction fragment polymorphisms a total of seven alleles could be distinguished. The allele frequencies were determined in six pairs of strains derived from different genetic base populations. Each pair consisted of two strains which differed in their susceptibility to Marek's disease (MD), a virus-induced neoplastic disease. The frequency of the most common haplotype (M2) was consistently higher in the susceptible strains than in the corresponding resistant strains (P < 0.05, Wilcoxon signed-ranks test), indicating that the observed differences were not due to random genetic drift. This result suggests that PEPCK-M may be a candidate gene which contains genetic variants affecting MD susceptibility. Variations in gluconeogenesis may affect the interplay between proliferation of neoplasia and host metabolism.

Alleles↗
PubMed1998

Association of endogenous viral genes with quantitative traits in chickens selected for high egg production and susceptibility or resistance to Marek's disease.

1. The association of endogenous viral (ev) genes with quantitative traits in 2 genetically distinct sets of White Leghorn strains were investigated. Strain S had been selected for susceptibility to Marek's disease (MD) whereas strain K had been selected for resistance to MD and high egg production and egg weight. 2. In all, 8 ev genes were typed. Ev10, ev19 and 'newB' occurred exclusively in strain S, 'newA' occurred only in strain K, and ev1, ev3, ev6 and ev8 occurred in both strains. 3. Whereas ev6 and ev8 were associated with reduction in egg production rate in strain S, in strain K, the presence of ev3 was associated with increased group specific antigen. 4. The genetic background of the chicken strain may play a role in the way certain ev genes affect traits. 5. It was thought that the position of ev genes on the chromosome may be important and their association with traits of economic importance make them potential genetic markers for uncovering quantitative trait loci.

Animals↗
PubMed1998

Genetic variability in white leghorns revealed by chicken liver expressed sequence tags.

A total of 92 expressed sequence tags from chicken liver (CLEST) were searched for homology with known genes. Among the CLEST, 29% had no sequence similarities with known genes, 34% showed sequence similarity to rRNA, 9% to mitochondrial genes, 23% to known nuclear genes, and 5% to human expressed sequence tags. Among the nuclear CLEST (excluding rRNA), clones with sequence similarity to aldolase B were represented four times, whereas all the other clones represented unique genes. The presence of MspI and TaqI restriction fragment length polymorphisms (RFLP) associated with CLEST were analyzed by bulk Southern blotting in 16 strains of White Leghorn chickens derived from five different genetic bases. No RFLP were observed with rRNA CLEST and a single MspI RFLP was observed with mitochondrial CLEST. The nuclear CLEST with sequence similarity to known nuclear genes were grouped into two classes on the basis of their involvement in intermediary metabolism. Among the nine genes coding for metabolic enzymes, all but one were polymorphic at MspI and/or TaqI sites in at least one of the strains, whereas among the other genes six of nine were polymorphic. The average frequency of clones revealing RFLP per cDNA clone and restriction enzyme for the two classes were 0.7 and 0.3, respectively. The analysis indicated that in White Leghorns, RFLP markers in the vicinity of nuclear CLEST are relatively frequent. Further, RFLP in the vicinity of genes coding for metabolic enzymes were significantly more frequent than near genes coding for other proteins.

Animals↗
PubMed1997

Changes in expression of the prolactin and growth hormone gene during different reproductive stages in the pituitary gland of turkeys.

The changes in levels of prolactin (PRL) and growth hormone (GH) in plasma and the pituitary gland and their transcripts were measured in turkey hens at different physiological stages by radioimmunoassay and dot blot hybridization analysis, respectively. The levels of tPRL mRNA in the pituitary gland increased from those of the immature group to the egg-laying group, reaching a maximum during the incubation and a minimum during the moulting stages. Changes in pituitary levels of PRL and PRL mRNA followed a similar trend and consequently were highly correlated (r2 = 0.83), whereas a significant but lower correlation was observed between circulating and pituitary levels (r2 = 0.62). Less significant changes were measured for tGH mRNA, with maximum and minimum levels measured in the pituitaries of egg-laying and non-laying hens, respectively. These data suggest that although changes in concentration of PRL are correlated with the reproductive stage of the turkey hen, coordinate changes in levels of GH are not.

Animals↗
PubMed1997

Trait association of genetic markers in the growth hormone and the growth hormone receptor gene in a White Leghorn strain.

Alleles of the growth hormone (GH) gene and GH receptor (GHR) gene were analyzed for association with juvenile body weight (HBWT), age at first egg (AFE), the hen-day rate of egg production (HDR), egg specific gravity (SPG), and egg weight (EWT) in a strain of White Leghorns. The particular strain segregated at near equal frequencies for two GH alleles defined by differences at three restriction fragment length polymorphisms (RFLP) and for two GHR alleles defined by a single RFLP. The GH genotype was significantly associated with AFE (P < or = 0.04) as well as HDR from 274 to 385 d (P < or = 0.04) and 386 to 497 d (P < or = 0.0003). The GHR genotype (haploid in female chickens) had trends for association with HBW (P < or = 0.06) and HDR from AFE to 273 d (P < or = 0.07). The effects on the egg quality traits SPG and EWT were not significant. Regression analysis revealed that HDR was associated negatively with AFE and positively with HBWT. The slope of the regression line of HDR on AFE varied with the GH genotype, with the effect that the differences in HDR between GH genotypes was relatively small in chickens with early AFE and large in chickens with late AFE. Similarly, the slope of the regression of HDR on HBWT varied between GHR genotypes, with the result that the effect of the GHR genotype on HDR in chickens with low HBWT was opposite to its effect in chickens with high HBWT. The complex relationship between genotypes and traits may reflect gene interaction and indicates that simple models based on additive gene effects may not be adequate for the dissection of the genetic architecture of quantitative traits.

Aging↗
PubMed1997

DNA polymorphisms in the chicken growth hormone gene: response to selection for disease resistance and association with egg production.

Analysis of the growth hormone (GH) gene in 12 strains of White Leghorn chickens revealed restriction fragment length polymorphisms (RFLPs) at three MspI sites and at a SacI site. Based on linkage disequilibrium analysis, they gave rise to eight different alleles (i.e. combinations of RFLPs), with five occurring at frequencies above 5% in at least one strain. Pairs of GH-RFLPs were at near maximal linkage disequilibrium, suggesting either a lack of recombination or the presence of selection pressure during evolution of the GH gene. Allele frequencies were determined in 12 non-inbred strains derived from three different genetic bases. These strains had been selected either for an array of egg production traits, resistance to Marek's disease or resistance to avian leukosis. Selection for disease resistance was consistently correlated with an increase in the frequency of one of the alleles. One strain segregated for only two alleles, which differed by three RFLPs. Analysis of variance in this strain indicated that the GH allele co-selected with resistance was associated with a delayed onset of ovulation but a higher persistency of ovulation as age progressed, resulting in an overall increase of egg production by 15% (age at first egg to 497 days). The resistance-associated GH allele was dominant for the onset of ovulation and recessive for the persistency of egg production. There was no significant effect of the GH genotype on juvenile body weight, egg weight or egg specific gravity.

Animals↗
PubMed1996

Molecular cloning, tissue distribution, and expression of the prolactin receptor during various reproductive states in Meleagris gallopavo.

The primary sequence of the prolactin receptor (PRL-R) in turkeys was deduced from a cDNA clone isolated from a kidney cDNA library and from a polymerase chain reaction (PCR) product. The open reading frame of the turkey PRL-R (tPRL-R) predicted an 831-amino acid protein composed of a leader peptide, an extracellular domain, a single transmembrane domain, and an intracellular domain. The extracellular domain contained two homologous repeat units with 63% amino acid sequence identity to each other. Each repeat unit contained all of the conserved cysteine pairs and a WSXWS motif found in mammalian PRL-Rs. A tPRL-R transcript with a molecular size of about 3000 nucleotides was identified by Northern blot analysis. The tPRL-R transcripts were detected in all 26 tissues examined using reverse transcriptase PCR (RT-PCR). The pituitary gland, hypothalamus, crop sac, duodenum, and gizzard were found to express the highest levels of tPRL-R among the 26 tissues. The expression levels of tPRL-R in 17 tissues were compared using semi-quantitative RT-PCR in nonphotostimulated, laying, out-of-lay, incubating, and maternal hens, and male birds. In most tissues examined there was no obvious relationship between blood levels of PRL, reproductive states, and estimated concentrations of the receptor mRNA. In the pituitary gland and hypothalamus, plasma levels of PRL and levels of tPRL-R transcript were inversely correlated. In the hypothalamus, increasing blood levels of PRL were associated with decreasing levels of the receptor transcript (p < or = 0.05), whereas the opposite was observed in the pituitary gland (p < or = 0.05). These findings support the hypothesis that PRL itself may participate in the neuroendocrine control of incubation behavior through actions on both the hypothalamus via a short-loop feedback mechanism and the pituitary gland via autocrine and/or paracrine effects.

Amino Acid Sequence↗
PubMed1995

Presence of an additional PstI fragment in intron 1 of the chicken growth hormone-encoding gene.

A previously unreported 196-bp PstI fragment was found in intron 1 of the gene encoding chicken growth hormone (cGH) when a PCR assay for an MspI restriction fragment length polymorphism was established. A pair of PCR primers was designed according to the published cGH sequence and used to amplify a fragment which contained two MspI sites, one polymorphic and another non-polymorphic. However, amplification of genomic DNA from two strains of meat-type chickens and three strains of White Leghorn chickens yielded a PCR product which was about 200 bp larger than expected. The fragment from one of the meat-type chickens was subcloned into the vector pCR-Script SK+, and sequenced. It revealed the presence of an extra fragment of 196 bp which was flanked by the PstI sites and occurred at nt +308 of the previously reported cGH sequence.

Animals↗
PubMed1994

Development and validation of a homologous radioimmunoassay using a biologically active recombinant turkey prolactin.

1. A new homologous radioimmunoassay has been developed for the measurement of turkey prolactin. 2. A 25000 kDa purified recombinant derived turkey prolactin (rtPRL), the biological activity of which was tested using a crop sac assay, was used as immunogen for the production of rabbit antiserum. In this biological test, the rtPRL was as active as the ovinePRL. 3. The radioligand (rtPRL) was labelled with 125I and the assay allowed the detection of standard doses of rtPRL ranging from 400 pg/tube to 50 ng/tube. 4. No cross reaction with chicken luteinising hormone and recombinant chicken growth hormone was detected. 5. The within and between assay coefficients of variability were 5.0 +/- 2.7% and 16.3%, respectively. The overall mean recovery ratio was 1.01. 6. The dose-response curves obtained with serial dilution of plasma and pituitary from turkey hens at different physiological stages and from male turkeys were parallel to those obtained with standard rtPRL. 7. The measured concentration of prolactin was 5 times higher in plasma from incubating than laying turkey hens, and the pituitaries from incubating hens contained 2 and 4 times more prolactin than those of laying and out of lay hens or males, respectively. 8. To further assess the validity of the assay, we measured changes in plasma concentration of prolactin in turkeys following stimulation with chicken vasointestinal peptide (cVIP). A single injection of 1 or 10 micrograms/kg body weight of cVIP to laying hens produced a large and rapid increase in plasma prolactin. 9. This new radioimmunoassay appears to be high for the measurement of turkey prolactin.

Analysis of Variance↗
PubMed1993

Synergism between the endogenous viral loci ev6 and ev9 in inducing immunological tolerance to avian leukosis virus.

1. The course of infection by exogenous avian leukosis virus was followed in a commercial strain of White Leghorn domestic fowls by measuring viral antigen in feather pulp and egg albumin. Ten days after hatching, 2 out of 360 birds tested positive and at 286 days of age about 60% of the birds had been antigen positive at least once. 2. Among the antigen positive birds, two groups could be distinguished: those which permanently and those which transiently expressed viral antigen. Permanent antigen expression was associated with low antibody titres, while transient antigen expression was associated with high antibody titres. 3. The strain segregated for the two endogenous viral genes ev6 and ev9, both of which express endogenous viral envelope protein, and have been implicated in affecting immune-responsiveness. The antibody titre in individuals positive for both ev6 and ev9, was significantly lower than in those which had none or only one of the two ev-genes. In addition, individuals positive for both ev-genes occurred more frequently in the group permanently positive for viral antigen than in the group transiently antigen positive. 4. The results indicate that there was a strong synergism between ev6 and ev9 in reducing the antibody response to exogenous avian leukosis virus infection, perhaps by inducing immune tolerance or interfering with antibody formation.

Animals↗