Expression of porcine FSHbeta subunit promoter-driven herpes simplex virus thymidine kinase gene in transgenic rats.
A transgenic rat was established using the construct of porcine FSHbeta subunit promoter, the -852/+10 bp region, fused to a Herpes simplex virus thymidine kinase (HSV-TK) gene. Integration of the transgene was confirmed by PCR of tail DNA. RT-PCR of total RNAs of the pituitary, gonad, cerebellum, liver, kidney, adrenal gland, prostate, and uterus revealed that FSHbeta was only expressed in the pituitary. Analysis of the expression of reporter gene, HSV-TK, using two specific primer sets revealed that different transcripts were present in the pituitary and testis. The transcript initiated at the transcription initiation site of the porcine FSHbeta gene was detected in the pituitary, and another within the TK gene was found in the testis, indicating ectopic testis-specific expression. Immunohistochemistry of the pituitary glands of the transgenic rats for FSH and HSV-TK demonstrated that the FSH-producing cells also produced HSV-TK. The results indicated that the -852/+10 bp region of the FSHbeta promoter contains an element(s) that determines the tissue-specific expression. We succeeded in producing FSHbeta promoter-driven HSV-TK transgenic rats and were the first time to do so using an animal other than the mouse. The transgenic rats show male infertility that involves abnormal spermatogenesis. We also observed a decrease in the weight of the testis and epididymis, and both motile and living spermatozoa were absent in the epididymis. Consequently, the FSHbeta-HSV-TK transgenic rat will provide a useful model for studies on FSH function and male infertility.
Cai
Li-yi
LY
Division of Life Science, Graduate School of Agriculture, Meiji University, Tama, Kawasaki, Japan.
Kato
Takako
T
Ito
Kazumi
K
Nakayama
Michie
M
Susa
Takao
T
Aikawa
Satoko
S
Maeda
Kei-Ichiro
K
Tsukamura
Hiroko
H
Ohta
Akihiko
A
Izumi
Shun-ichiro
S
Kato
Yukio
Y
D000818:Animals; D030801:Animals, Genetically Modified; D005260:Female; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006561:Herpes Simplex / Q000235:genetics*; D007248:Infertility, Male / Q000235:genetics; D008297:Male; D009929:Organ Size / Q000235:genetics; D010902:Pituitary Gland / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D013081:Sperm Motility; D013091:Spermatogenesis / Q000235:genetics; D013094:Spermatozoa / Q000473:pathology; D013552:Swine / Q000235:genetics; D013737:Testis / Q000473:pathology / Q000502:physiology; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism
D037201:Follicle Stimulating Hormone, beta Subunit; D013937:Thymidine Kinase