It is estimated that approximately 30% of men have reduced fertility and 2% are totally infertile. Despite these large numbers relatively little is know about the molecular bases of male infertility. On the flip side of male infertility is the need for male contraception. Currently there are no reversible, convenient male contraceptives available. In order to develop male contraceptives and acquire a greater understanding of male fertility there is a need to develop animal models to study the molecular basis and pathways that regulate and control male fertility. Vanderbilt researchers have developed a model mouse system to study male fertility. There research focuses on the epididymus, which is the area that spermatozoa acquire the ability to move and fertilize. For this region to be functional tissue and cell specific gene regulation must occur. These investigators have discovered one such gene regulated within this area, mEP17. These researchers can fuse either mouse or human EP17 or just the regulatory regions of either EP17 to reporter genes and the resulting fusion can be used to screen for substances that regulate this gene and affect male fertility. This system becomes a powerful tool to identify drugs which affect this gene and be potential male contraceptives. In addition polypeptides generated to this gene may be used as vaccines for male contraceptives.
We produced a plasmid containing the Fc portion of mouse IgGl (Fc) coupled to human fibroblast growth factor 1 (FGF-1). The plasmid was transformed into E. coli to express the fusion protein. The fusion protein was purified on a heparin sepharose column which has high affinity for the FGF portion of the fusion protein. The purpose of making this protein was to be able to identify cells that express receptors for FGF using flow cytometry.There are multiple fluorochrome labeled antibodies to mouse IgGl. When the fusion protein is bound to FGF receptors on cells, the Fc portion is on the surface of the cells and can be detected by fluorochrome labeled antibodies to mouse IgGl. Therefore, cells that express FGF receptors and bind the fusion protein can be detected by flow cytometry or immunofluorescence.
The invention is a cell line (Human embryonic kidney 293) stably expressing a recombinant human chloride channel (hClC-4). The cells enable high throughput screening of compounds that modulate chloride channel activity.
The invention is a cell line (Human embryonic kidney 293) stably expressing a recombinant human skeletal muscle voltage-gated sodium channel (hSkMl). The cells enable high throughput screening of compounds that modulate sodium channel activity.
The invention is a cell line (Human embryonic kidney 293) stably expressing a recombinant human skeletal muscle chloride channel (hClC-1). The cells enable high throughput screening of compounds that modulate chloride channel activity.
A method to assess oxidative stress in vivo includes the steps of measuring an amount of neuroprostanes in a biological sample before the ex vivo development of neuroprostanes in a sample, comparing the measured amount of neuroprostanes with a control and assessing oxidative stress in vivo based on this comparison. There is also provided a marker for oxidated stress by an increase of neuroprostanes in a biological sample compared to a control sample. A diagnostic tool for determining the presence of a neurodegenerative disease provides for determining an increased amount of neuroprostanes in a biological sample compared to that of a control sample.
This technology describes a novel methodology wherein human-induced pluripotent stem cells can be differentiated into neural precuror cells using DMH1 (a dorsomorphin analog). Neural progenitor cells can then be further differentiated into tyrosine hydroxylase expressing neurons.