Ovarian tissue excised from a Himalayan tahr (Hemitragus jemlahicus) served as the source from which the HJ1.Ov cell line was initiated. The Himalayan tahr is a relative of the wild goat native to the southern slopes of the Himalayas that has been introduced to New Zealand and other areas, where it tends to have a negative impact on native flora and fauna.

The HJ1.Ov line was established by The Naval Biosciences Laboratory (NBL) located in Oakland, California. The cells are epithelial in morphology and exhibit adherent growth to glass and polymer surfaces in monolayer culture. Similar to other epithelial cells, HJ1.Ov cells experience significant contact inhibition of migration.

Epithelial cells are a major class of cells that cover the body's surface and line its organs and cavities. In vivo, the cells are generally organized into contiguous sheets (epithelia) that primarily serve as diffusion barriers. The epithelial tissue that encases the ovaries is commonly called the germinal epithelium and is continuous with the peritoneum. The cells that comprise the germinal epithelium are capable of multiplying in significant numbers when needed, a characteristic that is imperative for species that undergo notable ovarian expansion during breeding season.

The HJ1.Ov epithelial cells that appear in the digital image above were resident in a cell culture that was stained with MitoTracker Red CMXRos, Alexa Fluor 488 conjugated to phalloidin, and Hoechst 33342, which target the mitochondrial network, filamentous actin, and DNA in the cell nucleus, respectively. Images were recorded in grayscale with a 12-bit digital camera coupled to either a Nikon E-600 or Eclipse 80i microscope equipped with bandpass emission fluorescence filter optical blocks. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.

Proximity of the Golgi Complex and the Nucleus in HJ1.Ov Cells - In a double immunofluorescence experiment, an adherent monolayer culture of tahr ovary epithelial cells was fixed, permeabilized, blocked with 10-percent normal goat serum, and then treated with a cocktail of mouse anti-NPCP (nuclear pore complex protein) and rabbit anti-giantin (Golgi complex) primary antibodies followed by goat anti-mouse and anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 568 and Alexa Fluor 488, respectively.

Tahr Ovary Cells Labeled for Keratin Intermediate Filaments - The log phase culture of HJ1.Ov cells illustrated in this section was treated for one hour with MitoTracker Red CMXRos in order to label the mitochondrial network, and the fixed cells were then incubated with mouse anti-cytokeratin primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Marina Blue. The nuclei were counterstained with SYTOX Green.

Visualizing Mitochondria, Filamentous Actin, and the Nucleus in Tahr Ovary Epithelial Cells - To target the mitochondrial network, cultured tahr ovary cells were treated with MitoTracker Red CMXRos in growth medium for one hour, washed, and fixed with 3.7-percent paraformaldehyde in medium containing serum. After washing and permeabilization, the cells were blocked with bovine serum albumen in PBS and labeled with BODIPY FL conjugated to phallacidin. The nuclei were subsequently counterstained with Hoechst 33258.

Distribution of Histone and Peroxisomal Membrane Proteins in HJ1.Ov Cell Cultures - The proximity of nuclei and peroxisomes in the tahr ovary cell culture illustrated in this section was probed in a double immunofluorescence experiment with mouse anti-histones (pan) and rabbit anti-PMP 70 (peroxisomal membrane protein) primary antibodies. The antibody targets were visualized with goat secondary antibodies conjugated to Alexa Fluor 568 and Alexa Fluor 488, respectively, while the actin cytoskeletal framework was labeled with Alexa Fluor 350 conjugated to phalloidin.

Triple Fluorophore Labeling of Ovarian Cells with MitoTracker Deep Red 633, Alexa Fluor 568, and YO-PRO-1 - The featured culture of tahr ovary epithelial cells was treated with MitoTracker Deep Red 633 and Alexa Fluor 568 conjugated to phalloidin, fluorescently labeling the mitochondrial network and F-actin, respectively. The nucleic acid stain YO-PRO-1 was utilized to counterstain cell nuclei.

Targeting Giantin and Nuclear Pore Complex Proteins in HJ1.Ov Cultures with Immunofluorescence - A culture of HJ1.OV epithelial cells was immunofluorescently labeled for the nuclear pore and Golgi complexes with a cocktail of mouse anti-NPCP (nuclear pore complex protein) and rabbit anti-giantin primary antibodies followed by goat anti-mouse and anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 488 and Alexa Fluor 568, respectively. The actin cytoskeleton was targeted with Alexa Fluor 350 conjugated to phalloidin.

Alexa Fluor Probes with Ultraviolet, Blue, and Green Excitation in Tahr Ovary Epithelial Cells - Employing a combination of ultraviolet, blue, and green excitation Alexa Fluor dyes, a monolayer culture of HJ1.Ov cells was triple-labeled using double immunofluorescence and a phallotoxin. Nuclei were visualized with mouse anti-NPCP (nuclear pore complex protein) primary antibodies, while the Golgi complex was stained with rabbit anti-giantin antibodies. Secondary antibodies were goat anti-mouse and anti-rabbit conjugated to Alexa Fluor 488 and 568. The filamentous actin network was counterstained with Alexa Fluor 350 conjugated to phalloidin.

Distribution of Golgi Bodies and Nuclear Pore Complex Proteins in Ovarian Cell Cultures - The HJ1.Ov cell culture featured here was immunofluorescently labeled with rabbit primary antibodies to giantin, a protein resident in the Golgi complex of mammalian cells. The culture was subsequently stained with a mixture of secondary antibodies conjugated to Alexa Fluor 568. In addition, nuclear pore complexes were immunofluorescently labeled with primary anti-NPCP mouse monoclonal antibodies followed by goat anti-mouse Fab heavy and light chain fragments conjugated to Alexa Fluor 488.

Localizing Filamentous Actin, Mitochondria, and the Nucleus in HJ1.Ov Cells - Triple fluorophore labeling was applied to the tahr ovary epithelial cells illustrated in this section. The relationship between the mitochondrial and filamentous actin networks was visualized with MitoTracker Red CMXRos and Oregon Green 488 conjugated to phalloidin, DNA in the cell nucleus was targeted with Hoechst 33258.

Tahr Ovarian Cell Culture Labeled for Histones and Peroxisomes with Immunofluorescence - Nuclear histone proteins were targeted in a culture of tahr ovary (HJ1.Ov) cells with mouse anti-histone (pan) monoclonal antibodies, which were imaged with goat anti-mouse Fab fragments conjugated to Alexa Fluor 568 (labeling the nucleus). The specimen was simultaneously labeled for peroxisomes with Alexa Fluor 488 conjugated to goat secondary antibodies that target rabbit anti-PMP 70.

The Vimentin Intermediate Filament and Filamentous Actin Networks in HJ1.Ov Cells - The proximity of intermediate filaments and the cytoskeletal filamentous actin network was visualized by treating the fixed and permeabilized culture of HJ1.Ov cells shown in the digital image above with mouse anti-vimentin primary antibodies followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 568. F-actin was subsequently labeled with Alexa Fluor 633 conjugated to phalloidin, and cell nuclei were counterstained with SYTOX Green.