Recent studies suggest that IGF-I is a crucial regulatory factor in follicular growth during early post-partum period. The aim of the present study was to determine in detail the changing profiles of metabolic and reproductive hormones in relation to ovulation of the dominant follicle (DF) of the first follicular wave post-partum in high-producing dairy cows. Plasma concentrations of related hormones in 22 multiparous Holstein cows were measured from 4 weeks pre-partum to 3 weeks post-partum, and the development of DF was observed with colour Doppler ultrasound. Thirteen cows showed ovulation by 15.2 days post-partum. Anovulatory cows showed higher GH and lower IGF-I levels than those in ovulatory cows during the peri-partum period. Each DF developed similarly, and a clear blood flow in the follicle wall was observed despite ovulation or anovulation. In addition, detailed endocrine profiles were analyzed in 9 out of the 22 cows. Five cows showed an increase in plasma oestradiol-17b (E2) with follicular growth followed by E2 peak, LH surge and ovulation. In these cows, plasma IGF-I concentrations remained high until 10 days post-partum followed by a gradual decrease. Subsequently, the insulin level increased together with the E2 peak towards ovulation. These profiles were not observed in anovulatory cows. In conclusion, our data strongly support the concept that IGF-I and insulin represent 'metabolic signals' of the resumption of ovarian function post-partum in high-producing dairy cows. Moreover, we provide the first visual evidence that both ovulatory and anovulatory DFs of the first follicular wave post-partum are similarly supplied with active blood flow.

This study was conducted to estimate the effects of kisspeptin-10 on blood concentrations of LH and GH in prepubertal dairy heifers. Heifers received a single injection of 1 mg kisspeptin-10 (nZ5) or saline (nZ5) intravenously, and serial blood samples were collected at 15-min intervals to analyze the response curves of both LH and GH after injection. Peakshaped responses were observed for concentrations of LH and GH, and the peaks were observed at 27G3 and 75G9 min, respectively, after injection, only in heifers injected with kisspeptin-10.

ABSTRACT. The relationship among nutritional status, systemic insulin-like growth factor-I (IGF-I) and ovarian function early postpartum were investigated. A total of 27 Holstein-Friesian cows, 10 that cycled normally within 20 days postpartum, 5 diagnosed with follicular cysts, 8 with persistent corpus luteum (CL) after the first ovulation postpartum and 4 with inactive ovaries were used for the study. Blood samples were collected 1-3 times per week, for 60 days pre-and postpartum, for IGF-I, progesterone, estradiol, free fatty acids (FFA), blood urea nitrogen (BUN), and aspartate aminotransferase (AST) determination. Inactive ovary and cystic cows had a higher body condition score before calving and lost more condition than normal or persistent CL cows. Immediately postpartum, IGF-I levels were higher and rose sharply in cows that cycled normally than in cystic, inactive ovary or persistent CL cows. At calving and early postpartum, FFA was higher in inactive ovary and cystic than in normal and persistent CL cows. There was a significant strong positive relationship between IGF-I and BUN, and strong negative relationships between IGF-I and FFA and AST in all groups. There was a positive relationship between serum IGF-I and estradiol in normal cystic and inactive ovary cows. This study found that overconditioned cows during the dry period or at calving, lost more body condition postpartum. These cows also had a deeper and longer period of negative energy balance (NEB), poor liver function and low circulating IGF-I concentrations early postpartum. Such cows were likely to have poor reproductive function as seen in development of cystic ovaries, persistent CL and inactive ovary. Changes in serum IGF-I early postpartum may help predict both nutritional and reproductive status in dairy cattle.

Abstract. In cows, interferon-tau (IFNT) regulates maternal recognition around days 15-19 after artificial insemination (AI). The present study hypothesized that if key target genes of IFNT are clearly upregulated in earlier stages of pregnancy, these genes could be use as indices of future pregnancy in cows. Therefore, we determined the expression of these genes in peripheral blood mononuclear leukocytes (PBMCs) and polymorphonuclear granulocytes (PMNs) during the maternal recognition period (MRP). Twenty multiparous Holstein cows were subjected to AI on day 0 and categorized into the following groups: pregnancy (Preg, n = 9), embryonic death (ED, n = 5) and non-pregnancy (NP, n = 6). Progesterone levels in the Preg group were higher than those in the NP group on days 12-21. ISG15 and OAS-1 (IFN-stimulated genes: ISGs) mRNA in PBMCs on day 8 was higher in the Preg group than in the NP group, and these mRNAs in PMNs was higher in the Preg group on day 5 than in the NP and ED groups. Interleukin-10 (IL-10, Th2 cytokine) mRNA expression increased on day 8 in the PBMCs of pregnant cows. Tumor necrosis factor α (TNFα, Th1 cytokine) mRNA expression was stable in all groups. In an in vitro cell culture experiment, IFNT stimulated mRNA expression of ISGs in both PBMCs and PMNs. IFNT stimulated IL-10 mRNA expression in PBMCs, whereas IFNT increased TNFα mRNA levels in PBMCs in vitro. The results suggest that ISGs and IL-10 could be responsive to IFNT before the MRP in peripheral blood immune cells and may be useful target genes for reliable indices of pregnancy before the MRP. Key words: Cow, Immune cells, Interferon tau, Pregnancy (J. Reprod. Dev. 58: [84][85][86][87][88][89][90] 2012) D uring the past 5 decades, milk production per cow has dramatically increased because of improved management, nutrition, and genetic selection [1,2]. In contrast, the decreases in fertility and conception rates of the modern high-producing dairy cow are the major causes of economic loss for dairy producers [2,3]. After establishment of pregnancy in domestic ruminants, the conceptus secretes interferon-tau (IFNT) as a maternal recognition factor [4]. IFNT acts in the uterus around day 16 after insemination and prevents luteolysis by inhibiting prostaglandin F 2α release, resulting in the maintenance of corpus luteum function; therefore, this period is termed the maternal recognition period (MRP) [5]. IFNT induces the synthesis and secretion of IFN-stimulated genes (ISGs) such as ISG15, 2′,5′-oligoadenylate synthetase (OAS-1), IFN regulatory factor 1, Mx1 and Mx2 not only in the uterus but also in blood cells in ewes and cows [6][7][8][9][10]. In fact, in bovine peripheral blood leukocytes, ISG15 mRNA levels were higher in pregnant cows than in nonpregnant cows on days 18 and 20 after artificial insemination (AI) [7][8][9]. Technology for the early detection of pregnancy is needed to identify nonpregnant cows and to synchronize and artificially inseminate these cows prior to the next ovulation, and many studies have focused on identify...

Active angiogenesis and progesterone (P) synthesis occur in parallel during development of the corpus luteum (CL). Basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) are known to stimulate angiogenesis and P synthesis in vitro. The aim of the present study was to investigate the impact of bFGF or VEGF on the CL development in the cow by using a specific antibody against bFGF or VEGF. bFGF antibody, VEGF antibody, or saline as a control (n = 4 cows/treatment) were injected directly into the CL immediately after ovulation (Day 1), and the treatment was continued for 3 times/day over 7 days. Luteal biopsies were applied on Day 8 of the estrous cycle to determine the expression of genes associated with P synthesis and angiogenesis. Intraluteal injections with the bFGF antibody or the VEGF antibody markedly decreased the CL volume, plasma P concentration and StAR mRNA expression. bFGF antibody treatment decreased the mRNA expression of bFGF, FGF receptor-1, VEGF120, and angiopoietin (ANPT)-1, and increased ANPT-2/ANPT-1 ratio. However, VEGF antibody treatment decreased ANPT-2 mRNA expression and ANPT-2/ANPT-1 ratio. These results indicate that local neutralization of bFGF or VEGF changes genes regulating angiogenesis and P synthesis, and remarkably suppresses the CL size and P secretion during the development of CL in the cow, supporting the concept that bFGF and VEGF control the CL formation and function.