We hypothesized that the ovulatory response of one ovary to FSH would be related positively to the size of the primordial and growing pools of follicles in the other ovary. Nonlactating cows (n = 26) were unilaterally ovariectomized and 2 days later were superovulated. The superovulatory response was classified as Low (< 5 corpora lutea [CL]), Medium (5-14 CL), or High (> 14 CL). Surface follicles on the ovary removed before superovulation were classified as small (1-3 mm), medium (3-7 mm), or large ( > 7 mm), and the ovary was then fixed and serially sectioned. Follicles = 1 mm in diameter in 388 +/- 38 fields (2 x 2 mm) per cow were classified as primordial, primary, secondary, or tertiary. By classification, Suboptimal ovaries contained < 100 follicles = 1 mm and Optimal ovaries contained > 250 follicles = 1 mm. Number of CL was correlated positively with total number of primordial, tertiary, and medium surface follicles. Number of Empty fields (2 x 2-mm fields containing no follicles) was correlated negatively with superovulatory response and number of primordial follicles. Number of CL was related to number of tertiary follicles in a positive linear manner and the number of medium follicles in a positive quadratic manner (r 2 = 0.66). Numbers of primordial, tertiary, small surface follicles, medium surface follicles, and total surface follicles were lower (p = 0.06) in the Low superovulatory response group than in the Medium or High group. Suboptimal ovaries had fewer small surface follicles and fewer CL than Optimal ovaries (p < 0.05). We conclude that superovulatory response in cattle is related positively to the pools of primordial and growing follicles in the bovine ovary.
Previous research revealed that treatment with vitamin A approximately 5 d before ovulation may increase litter size in weaned sows and improve embryonal survival in gilts fed high-energy diets that reduced embryonal survival. For the current study, the hypothesis was that administration of vitamin A before ovulation would alter development of follicles and oocytes in a way favorable to enhanced embryonal survival. (Landrace x Large White) x (Duroc x Hampshire) gilts (n = 44) were fed 11.0 Mcal ME x gilt(-1) x d(-1) beginning 7 d after second estrus and given (i.m.) corn oil or 1 x 10(6) IU of vitamin A (retinyl palmitate) on d 15 after second estrus. Gilts were checked for estrus every 4 h, mated naturally at third estrus, and assigned randomly to undergo midventral laparotomy beginning at 24 to 28, 28 to 32, 32 to 36, or 36 to 40 h after onset of third estrus. At laparotomy, ovulated oocytes and early-stage embryos were recovered from oviducts, and ovaries were removed for aspiration of oocytes and granulosa cells from unovulated follicles. Oocytes and embryos were stained for assessment of stage of development. Granulosa cells were cultured to assess their ability to secrete progesterone. Follicular fluid was assayed for progesterone, estradiol-17beta, IGF-I, and PGF2alpha. Treatment with vitamin A altered development of oocytes and embryos by decreasing the percentage at the germinal vesicle stage and increasing the percentage at advanced stages. Mean stage of development was increased by vitamin A, but variation in stage was decreased. Among follicles matched by meiotic stage of oocyte, follicular fluid concentrations of progesterone, IGF-I, and PGF2alpha were greater in vitamin A-treated gilts than in controls, but treatment with vitamin A in vivo did not affect LH-stimulated or unstimulated secretion of progesterone by granulosa cells in vitro. These data provide evidence that vitamin A may influence embryonic development by advancing resumption of meiosis and altering follicular hormonal environment during follicle maturation.
We hypothesized that the number of microscopic follicles present in the ovaries of cattle selected for twin births (Twinner) would be greater than in the ovaries of contemporary Controls. Ovaries were collected from seven Control and seven Twinner cows at slaughter. The number of Small (1 to 3.9 mm), Medium (4 to 7.9), and Large (> 8 mm) surface follicles was counted and one ovary was fixed for histological evaluation. Fifty to sixty consecutive 6-microm slices were taken from a piece of cortical tissue, approximately 1 cm x 1 cm in area, located between the surface follicles. Microscopic follicles were classified as primordial (oocyte surrounded by a single layer of squamous pregranulosa cells), primary (oocyte surrounded by a single layer of one or more cuboidal granulosa cells), secondary (oocyte surrounded by two or more layers of granulosa cells), or tertiary (oocyte surrounded by multiple layers of granulosa cells with initiation of antrum formation to < or = 1 mm in diameter). The total number of follicles was counted in 200 fields (2 mm x 2 mm) per ovary. A field containing no follicles was classified as empty. There were significantly more secondary follicles in Twinner compared with Control ovaries (12.9 vs 6.3; P < .05). Twinners also tended to have more small surface follicles (35.4 vs 49.0; P < 0.1). We conclude that ovaries of Control and Twinner cows do not differ in the number of primordial follicles or in the number of follicles activated into the growing pool; however, Twinner cows are able to maintain more growing follicles at the secondary and subsequent stages of development.
The hypothesis was that administration of vitamin A before ovulation would improve embryo survival in gilts fed a high-energy diet intentionally to reduce embryo survival. Forty crossbred ([Landrace x Large White] x [Duroc x Hampshire]) gilts were fed control (5.5 Mcal ME/d) or high-energy (11.0 Mcal ME/d) diets from 7 d after second estrus until 11 to 12 d after third estrus. Gilts in each dietary group received (i.m.) corn oil or retinyl palmitate (1 x 10(6) IU, vitamin A) on d 15 after second estrus and were mated at third estrus. Blood for determination of progesterone and estradiol was collected twice daily. The uterus and ovaries were removed on d 11 or 12 after third estrus for assessment of number of CL, and number, size and aromatase activity of embryos. Neither diet nor vitamin treatment affected number of CL. The high-energy diet exerted a negative effect on number of embryos (P = .09) and embryo survival (P = .07), whereas vitamin A exerted a positive effect on number of embryos (P = .07) and embryo survival (P = .08). The high-energy diet increased variation in embryo diameter, whereas vitamin A reduced variation in diameter and increased average diameter. Neither diet nor vitamin treatment influenced aromatase activity of embryos. Diet and vitamin treatment interacted with day to influence serum progesterone, but not estradiol. Injecting vitamin A before estrus restored embryo survival to normal levels in gilts fed high-energy diets, and this may be attributable to decreased variation in size of embryos.
Expression and localization of mRNAs for vascular endothelial growth factor (VEGF), VEGF receptor 1 (Flt) and VEGF receptor 2 (KDR) (VEGFR-1 and VEGFR-2, respectively) were investigated in pig corpora lutea. Northern blot analysis of total RNA indicated hybridization of pig VEGF, VEGFR-1 and VEGFR-2 cDNA probes to mRNA transcripts of approximately 3.9, 7.0 and 5.0 kb, respectively. The expression of mRNAs for VEGF and its receptors during the luteal phase (days 4, 7, 10, 13 and 15 after the onset of oestrus) were assessed by northern blot analysis, and hybridization signals were normalized to expression of pig 18S rRNA. Relative hybridization signals of expression of VEGF mRNA appeared to be constant; however, expression of VEGFR-1 mRNA was low on day 4, increased on day 7, and was higher on days 10, 13 and 15 (P<0.05, compared with day 4). In contrast, no changes in expression of mRNA for VEGFR-2 were evident on days 4-13, but a decrease was detected (P<0.05) at day 15. In situ hybridization revealed that VEGF mRNA was localized predominantly in large luteal cells, whereas both VEGFR-1 and VEGFR-2 were localized to small cells. These data indicate that the VEGF system may be involved in the regulation of luteal vasculature throughout the lifespan of the corpus luteum. Although the expression of VEGF mRNA was unchanged during the luteal phase, variations in the expression of VEGFR-1 and VEGFR-2 mRNAs indicate that differential regulation of expression of the VEGF receptors may play a role in the control of VEGF-mediated vascular growth at different phases of development and maturation of the pig corpus luteum.
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