Plasma Concentration of Estrone Sulfate during Pregnancy in Different Breeds of Japanese Beef Cattle

Nakao

2005

Animal Science Journal

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The present study was undertaken to develop a novel, practical and simple procedure for enzyme-linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti-estrone antibody and horseradish peroxidase-labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra-assay and inter-assay coefficients of variation were 5.3-8.1% and 13.4-15.7%, respectively. The recovery rate was 78-102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.

Section: Discussionsupporting

confidence: 88%

Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

Nakao

2005

Animal Science Journal

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“…The enzyme immunoassay procedure for progesterone and estradiol-17β were performed as described previously [17,[19][20][21][22][23]. The antibody for progesterone was raised with rabbit [19] and that for estradiol-17β was purchased from Cosmo Bio (Tokyo, Japan).…”

Section: Enzyme Immunoassay For Estradiol-17β and Progesteronementioning

confidence: 99%

Deficient Proliferation and Apoptosis in the Granulosa and Theca Interna Cells of the Bovine Cystic Follicle

Journal of Reproduction and Development

Yoshimura

2007

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Abstract. The aim of the present study was to examine the frequencies of cell proliferation and death of granulosa and theca interna layers during development of cystic follicles in order to understand the mechanisms of cystic follicle formation. Paraffin sections of cystic follicles were immunostained with antibodies against proliferating cell nuclear antigen (PCNA) and cleaved caspase-3 in order to observe proliferating and apoptotic cells, respectively. The concentrations of estradiol-17β and progesterone in the follicular fluid of these follicles were measured by ELISA. The granulosa and theca interna layers contained both PCNA-and caspase-3-positive cells, although their numbers were limited. There was significant negative correlation between the estradiol-17β and progesterone concentrations in the follicular fluid. Regression analysis revealed no significant correlation, except for that between the PCNA-positive cells in the theca interna and the caspase-3-positive cells in the granulosa layer. These results indicate that the granulosa and theca interna cells of the cystic follicle show weak proliferative activity and low apoptotic frequency; this implies that the cystic follicle grows slowly and then maintains a static condition without degeneration, which leads to long-term persistence of the follicle.

“…The estimated age of SB was 5-9 years, determined by dental examination [27]. Blood samples were also collected from SB before slaughter for determination of plasma progesterone level [28][29][30] to estimate whether the animals were in the luteal phase (>1 ng/ml) or not. However, age and progesterone concentration of HF were not i n v e s t i g a t e d d u e t o r e s t r i c t i o n s i n t h e slaughterhouse.…”

Section: Collection and Preparation Of Tissuementioning

confidence: 99%

Immunolocalization of von Willebrand Factor and Vascular Endothelial Growth Factor during Follicular Atresia in the Swamp Buffalo Ovary

Feranil

Journal of Reproduction and Development

Nakao

2005

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Abstract. The aim of this study was to investigate the distribution pattern of von Willebrand factor (vWF) and vascular endothelial growth factor (VEGF) in the healthy antral and atretic follicles of Philippine swamp buffaloes (SB) in comparison with Holstein-Friesian cows (HF). Paraffin sections of healthy follicles and atretic follicles at various stages were immunostained with vWF antibody and VEGF antibody. The density of vWF-positive capillary vessels in the theca interna significantly increased as atresia progressed in SB, whereas the density significantly decreased in late atretic follicles compared with advanced ones in HF. On the other hand, the area of vWF-positive capillary vessels in the theca interna significantly increased as atresia progressed in both SB and HF. Immunoreactions of VEGF in the granulosa cells (in all follicle types) were observed in both SB and HF. In the granulosa layer, a reduction in the VEGF immunoreaction was noted as follicles progressed from healthy to advanced atretic follicles in both animals. Granulosa cells (in both SB and HF) showed a higher immunopositive staining than theca cells. In the theca interna, VEGF immunostaining diminished as follicles progressed to the late atretic follicles in both animals. These results indicate that during atresia, changes of vWF expression are the opposite of VEGF expression in SB. Both vWF and VEGF are suggested to be associated with follicular atresia in SB.