Biomedical subjects

Tim King

Publications and source records attributed to Tim King.

9 recordsLinked to original sources

PubMed2006

Maintenance of pregnancy in pigs with limited viable embryos.

Interest is increasing in assisted reproductive technologies in the pig involving embryo cryopreservation, cloning, and genetic modification. Although inherently inefficient and variable in their outcome, the successful application of these techniques in this species is confounded by unique mechanisms for pregnancy recognition and maintenance that require a minimum number of viable embryos during a critical window of time early in gestation. These mechanisms require both local and systemic interactions between conceptuses and the maternal reproductive axis. Here, we describe a method wherein cotransfer of parthenogenetic embryos with the capacity for limited development can be used to sustain the pregnancy of fewer than four viable conceptuses to term.

Animals↗

PubMed2005

Cushing's syndrome due to adrenal carcinoma.

Cushing's syndrome results from chronic excessive endogenous or exogenous glucocorticosteroids. Extremely high concentrations cause an accelerated syndrome with rapid deterioration, often suggesting ectopic ACTH production. We present a case of adrenocortical carcinoma-causing corticotrophin-independent Cushing's syndrome.

Adrenal Gland Neoplasms↗

PubMed2005

Live lambs born from zona-pellucida denuded embryos.

Progress with techniques using zona-pellucida denuded embryos has resulted in the birth of live cattle, pigs, and mice. The application of zona-free methods in sheep has been restricted to in vitro studies. In this report, we demonstrate that live lambs can be produced from zona-free IVF embryos. We are pursuing this method as a prerequisite to developing viral vector co-culture delivery strategies.

Animals↗

PubMed2004

Mouse myotomes pairs exhibit left-right asymmetric expression of MLC3F and alpha-skeletal actin.

Most muscle originates from the myotomal compartment of the somites, paired structures flanking the neural tube. Whereas vertebrate embryos show molecular and morphological asymmetry about the left-right body axis, somitic myogenesis is thought to occur symmetrically. Here, we provide the first evidence that myotome pairs are transiently left-right asymmetric, with higher expression of alpha-skeletal actin and myosin light chain 3F (MLC3F) on the left side between embryonic day 9.5-10.25. In iv mutants with situs inversus, the asymmetric expression of alpha-skeletal actin and MLC3F was inverted, showing that this process is regulated by global left-right axis cues, initiated before gastrulation. However, although left-sided identity is later maintained by Pitx2 genes, we found that Pitx2c null embryos have normal left-biased expression of alpha-skeletal actin and MLC3F. Myotome asymmetry, therefore, is downstream of the iv mutation but upstream of, or unrelated to, the Pitx2c pathway.

Actins↗

PubMed2003

Application of reproductive biotechnology in animals: implications and potentials. Applications of reproductive cloning.

The development of new methods of nuclear transfer in mammals is creating many new opportunities in research, medicine and agriculture. The method of cloning is repeatable and has been established in many laboratories worldwide. However, the present procedure is inefficient with fewer than 4% of embryos becoming viable offspring. A considerable improvement in efficiency is required before wide scale use for livestock improvement. The opportunity to introduce precise genetic changes to livestock is available for the first time through the use of gene targeting procedures in cultured cells that are used as nuclear donors. This has potential application in the production of organs for transplantation to humans, studies of human genetic disease and basic research in to the control of gene expression and function.

Agriculture↗

PubMed2003

Telomerase-immortalized sheep fibroblasts can be reprogrammed by nuclear transfer to undergo early development.

Telomere shortening and lack of telomerase activity have been implicated in cellular senescence in human fibroblasts. Expression of the human telomerase catalytic reverse transcriptase subunit (hTERT) in these cells reconstitutes telomerase activity and immortalizes the cells without tumor transformation. In this report, we show that sheep fibroblasts are similar to human cells. They do not have detectable telomerase activity and undergo only a finite numbers of cell divisions before replicative senescence. Telomere lengths in sheep fibroblasts are similar to those reported for human cells and shorten at a rate of 50-200 base pairs (bp) each cell division. Expression of the human telomerase catalytic subunit restored the telomerase activity in the sheep cells and extended their proliferative life span. None of the telomerase positive sheep fibroblasts exhibited a transformed phenotype after 200 days of continuous culture, and the higher hTERT expressing cells maintained their telomere lengths and normal cell characteristics for more than 500 days in culture. In cloning experiments using one of these cell lines as a nuclear donor, the reconstructed karyoplasts were reprogrammed and developed to the blastocyst stage at a similar frequency to that observed with the parental, telomerase negative cell line. After embryo transfer the blastocysts exhibited a relatively high frequency of implantation, early fetal development, and organogenesis. No fetuses survived beyond 40 days of development, however, showing that although these cells could be substantially reprogrammed, they were not fully competent for nuclear transfer.

Animals↗

PubMed2003

In vitro and in vivo developmental competence of ovulated and in vitro matured porcine oocytes activated by electrical activation.

The objective of this study was to evaluate the in vitro and in vivo developmental competence of parthenogenetic (parthenote) pig embryos derived from ovulated and in vitro matured (IVM) oocytes. A total of four experiments were carried out. These demonstrated that the mean blastocyst rates from stimulated ovulated and IVM pig oocytes were not significantly different (61% vs. 46%, p > 0.05) following in vitro culture. Both ovulated and IVM pig parthenotes were able to develop in vivo for 30 days. Parthenote fetuses collected 21 and 30 days post estrus were morphologically normal but significantly smaller and lighter than fertilized controls (p < 0.01). IVM pig parthenotes stopped development around 31 days post estrus.

Animals↗

PubMed2002

Expression of Peg1 (Mest) in the developing mouse heart: involvement in trabeculation.

Peg1 (Mest) is an imprinted gene of unknown function widely expressed in the mouse embryo, predominantly in cells of the mesodermal lineage. We have revealed a restricted expression pattern within the developing heart. Initial uniform expression throughout the linear heart tube subsequently becomes restricted, primarily to the developing myocardial trabeculae of both the atria and ventricles, where it persists into late development. Expression in the atrial appendage myocardium precedes the emergence of trabeculae (pectinate muscles), and occurs earlier and to a greater extent on the right than on the left, reflecting the spatial and temporal pattern of trabeculation. Analysis of myocardial morphology in mice lacking the Peg1 gene, which are viable and appear grossly normal, reveals a subtle alteration in the pattern of trabeculation: an increase in thickness and reduction in density of the compact myocardium, similar to that seen in the human cardiomyopathy ventricular noncompaction.

Alleles↗

PubMed2002

Somatic cell nuclear transfer: recent progress and challenges.

Somatic cell nuclear transfer (NT) offers new and exciting opportunities in many areas of research and biotechnology. However, the field as a whole is still in its infancy, with continuing inefficiencies in the process proving many early expectations premature. The technical steps of NT are complex, and success is highly susceptible to minor variations. Furthermore, the biological process of reprogramming is not fully understood, making it difficult to optimize the protocols for providing ideal recipient oocytes and donor cells. In this paper, we describe recent advances and novel approaches, which resulted in progress during the last year, including the birth of cloned piglets and farm animals with precise genetic changes. Key problems hindering further progress are addressed.

Animals↗