Identification and expression analysis of connexin-45 and connexin-60 as major connexins in porcine oocytes.
During mammalian oogenesis, intercellular communication between oocytes and the surrounding follicle cells through gap junction channels is crucial for oocyte development and maturation. The channel properties of gap junctions may be affected by the composition or combination of connexins, the expression of which is regulated by gonadotropins and other factors. Thus, identification and expression analysis of connexin genes in oocytes and follicle cells will help us to better understand how oogenesis and folliculogenesis are regulated in a species-specific manner in mammals. We previously reported the spatiotemporal expression of multiple connexin genes in porcine follicle cells. Here, we searched for connexin genes specifically expressed in porcine oocytes that may be involved in the formation of gap junctions between oocytes and follicle cells. To achieve this, we constructed an oocyte-specific cDNA library to identify which connexin genes are expressed in these cells and found that gap junction protein, alpha 10, which encodes connexin-60, and a porcine ortholog of mouse gap junction protein, gamma 1 encoding connexin-45, are the major connexins expressed in porcine oocytes during folliculogenesis. Immunostaining and in situ hybridization of sectioned porcine ovaries confirmed oocyte expression of these genes at 3 different stages of ovary development. Furthermore, their gap junction channel activity was assessed using a heterologous cell system. However, gap junction protein, alpha 4, which encodes connexin-37 and is expressed in the oocytes of several other mammals, was undetectable. We demonstrate that there is diversity in the connexin genes expressed in mammalian oocytes, and hence in the gap junctions connecting oocytes and cumulus cells.
Nitta
M
M
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Yogo
K
K
Ohashi
M
M
Akiyama
M
M
Kunitomo
Y
Y
Ogawa
T
T
Ishida-Kitagawa
N
N
Miyoshi
J
J
Sato
E
E
Takeya
T
T
D000818:Animals; D003001:Cloning, Molecular; D017630:Connexins / Q000032:analysis* / Q000096:biosynthesis / Q000502:physiology; D005260:Female; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D020869:Gene Expression Profiling / Q000662:veterinary; D015723:Gene Library; D017403:In Situ Hybridization / Q000662:veterinary; D021141:Nucleic Acid Amplification Techniques / Q000662:veterinary; D009865:Oocytes / Q000737:chemistry* / Q000502:physiology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D013552:Swine / Q000502:physiology; D000097006:Gap Junction alpha-4 Protein
D017630:Connexins; D000097006:Gap Junction alpha-4 Protein; C082920:connexin 45