Synthesis and characterization of biologically active recombinant elk and horse FSH.

This research investigation focuses on the cloning and expression of the alpha-subunit and FSH beta-subunit cDNAs from elk and horse, and the production of recombinant FSH from these species in vitro. The study also explores the impact of these recombinant products on ovarian growth when injected into immature female Wistar rats.

Methodology

• The researchers extracted RNAs from the pituitary glands of elk and horse.
• These RNAs were then reverse-transcribed and amplified by a technique known as a polymerase chain reaction.
• The cDNAs or complementary DNAs corresponding to both subunits of FSH (Follicle Stimulating Hormone) in elk and horse were cloned.
• These cDNAs were then inserted into the expression vector pBudCE4.1 and transfected into CRL-9096 cells, a type of cell line.

Analysis of Gene Expression

• The expression of these genes was determined in the transfected cells through Northern and Western blot analysis, techniques used for detection of specific molecules in a sample.

Characterization of Recombinant FSH

• The recombinant FSH secreted in the culture media from these cells was then characterized using an in vitro bioassay and RIA (radioimmunoassay), a method used to measure concentrations of antibodies.
• The study found that the recombinant horse FSH was 5.6 times more potent than the elk FSH when assessed by their activity over mass.

Impact on Ovarian Growth

• These recombinant products were injected into immature female Wistar rats to observe the effect on ovarian growth.
• The injections stimulated ovarian growth in these rats, suggesting these products are, indeed, recombinant versions of the native elk and horse FSH.

Significance of the Study

• This study indicates that these recombinant products may be used to improve methods of ovarian stimulation in cervids, equids, and other species, contributing to more predictable and efficient techniques in the field of reproductive biology.