Electrophoretic Characterization of Bovine Lipoprotein Subfractions Isolated by Agarose Gel Chromatography

Ferreri, L. F.; Gleockler, D H

doi:10.3168/jds.s0022-0302(79)83464-5

Cited by 20 publications

(9 citation statements)

References 17 publications

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“…An HDL was identified in bovine OF. The density range at which this HDL was isolated corresponds to that reported by Ferreri and Gleockler (1979) and Stead and Welch (1975) for HDL from bovine plasma. After ultracentrifugation, slightly different flotation characteristics were observed for HDL from OF, bovine serum HDL, and rabbit plasma HDL.…”

Section: Discussionsupporting

confidence: 76%

“…Differences in physical and chemical characteristics between HDL from plasma and OF may be related to the origin of the HDL particle. OF is partially derived by selective transudation of plasma components (Leese, 1988), and HDL is the major class of lipoprotein found in bovine plasma, making up 83% of the total lipoproteins (Ferreri and Gleockler, 1979). HDL is the only class of lipoprotein present in bovine follicular fluid (Brantmeier et al, 1987) that is also partially derived from plasma.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Bovine oviductal fluid components and their potential role in sperm cholesterol efflux

Ehrenwald¹,

Foote²,

Parks³

1990

Molecular Reproduction Devel

127

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Bovine oviductal fluid (OF) was collected and analyzed throughout the estrous cycle, and the capacity of the protein and lipoprotein components to support cholesterol efflux from bovine sperm was evaluated. Blood was collected and assayed for progesterone (P4) to monitor the estrous cycle. Protein and lipoprotein separation was achieved by density gradient centrifugation. Two major bands were identified. The first (1.056 less than delta 20 less than 1.140 g/ml) corresponded to bovine and rabbit plasma high-density lipoprotein (HDL) based on distribution in the density gradient and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The second band (1.235 less than delta 20 less than 1.243 g/ml) consisted predominantly of oviductal fluid albumin (OFA). Oviductal fluid protein concentration increased as serum P4 decreased around the time of estrus. Mean OF protein concentration was 21.3 mg/ml when serum P4 was lower than 0.5 ng/ml and 6.9 mg/ml when serum P4 was greater than 0.5 ng/ml. An inverse log relationship was found between HDL protein concentration and serum P4. Unesterified cholesterol (UC), cholesteryl ester, and phospholipid (PL) content of HDL for HDL protein concentrations of 3-56.1 micrograms/ml were 1.35-46.2 micrograms/ml, 1.91-44.48 micrograms/ml, and 1.69-59.8 micrograms/ml, respectively. Phosphatidylcholine and -ethanolamine were the major PLs present in the HDL fraction and their molar ratio (4:1 mol/mol) was relatively constant through the estrous cycle. The OFA fraction of the same samples accounted for more than 90% of total protein and for most of the variation in OF protein. To determine the ability of OF components to serve as sperm cholesterol acceptors, OF samples were incubated 1:1 (v/v) with and without 4 X 10(8) bovine sperm in 1.0 ml of modified Tyrode's solution and OF for 2 hr at 39 degrees C. After incubation, HDL and OFA fractions were isolated and analyzed for changes in protein and lipid content. After OF, samples were incubated with sperm, an increase in UC was found in the HDL fractions. UC in HDL increased by 12.1 +/- 1.0 micrograms/ml (means +/- SE) when serum P4 was less than or equal to 0.5 ng/ml. For samples corresponding to higher serum P4, the increase in UC was 3.60 +/- 0.89 micrograms/ml. Values for UC in HDL were corrected for the contribution of UC from OFA of OF samples. Cholesterol efflux from sperm has been implicated in the process of sperm capacitation. These results indicate that HDL from OF is elevated during the follicular phase of the estrous cycle and can serve as an acceptor for bovine sperm cholesterol.

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Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Bovine oviductal fluid components and their potential role in sperm cholesterol efflux

Ehrenwald¹,

Foote²,

Parks³

1990

Molecular Reproduction Devel

127

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“…Analytical and preparative ultracentrifugation, electrophoresis, precipitation by sulfated polysaccharides, and gel filtration are techniques which have been used to isolate and characterize bovine lipoproteins (1)(2)(3)(4)(5)(6). Preparative ultracentrifugation has been used extensively to isolate bovine lipoproteins on the basis of their hydrated densities; however, this method kas several disadvantages.…”

Section: Introductionmentioning

confidence: 99%

Comparison of ultracentrifugation and gel filtration for the isolation of bovine lipoproteins

Grummer

Davis

Hegarty

1983

Lipids

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Lipoproteins from the plasma of three nonlactating Holstein cows were isolated using either preparative ultracentrifugation or gel filtration chromatography. Lipoprotein classes obtained by ultracentrifugation were very low density plus chylomicra, less than 1.006 g/ml; low density, 1.007-1.039 g/ml; high density1, 1.040-1.063 g/ml; and high density, 1.064-1.22 g/ml. These lipoprotein classes were individually applied to an agarose gel column to determine at what volume they eluted in comparison to lipoproteins that were separated after applying total bovine lipoproteins to the column. Three major peaks corresponding to very low density lipoproteins plus chylomicra, low density, and high density lipoproteins resulted after gel filtration of total lipoproteins. Very low density lipoproteins plus chylomicra, obtained by ultracentrifugation, eluted as a single peak, as did low density and high density lipoproteins. However, high density1 lipoproteins eluted as two peaks. The first peak eluted at the same volume as low density lipoproteins, and the second peak eluted at a volume similar to that of the ascending slope of the high density lipoprotein peak. Results from disc polyacrylamide gel electrophoresis, immunoelectrophoresis and double immunodiffusion of lipoprotein fractions, and SDS polyacrylamide gel electrophoresis of their apoproteins, similarly indicated that the lipoproteins present in the 1.040-1.063 g/ml density interval are a mixture of low and high density lipoproteins rather than a unique class of lipoproteins.

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“…Grummer et al [9] reported that three major peaks (TRL, LDL, and HDL) were observed on the elution profiles of TLP in both lactating and non-lactating cows by gel permeation chromatography, however, only two peaks (TRL and HDL) were observed in lactating cows in this study. Ferreri and Gleockler [6] also detected two peaks in lactating cows and speculated that the lack of LDL peak was resulted from large HDL particles eluted broadly at the same position in LDL. No significant difference of apoB concentrations in LDL between lactating cows and non-lactating cows observed in this study supported that LDL peak was involved in HDL peak on the elution profile.…”

Section: Discussionmentioning

confidence: 99%

“…To investigate bovine lipoproteins and lipid metabolism, ultracentrifugation, electrophoresis, and gel permeation chromatography techniques have been commonly used [6,15,16]. Among these techniques, gel permeation chromatography is useful for both the analysis and preparation of bovine lipoproteins.…”

Section: Discussionmentioning

confidence: 99%

Utilization of Intestinal Triglyceride-rich Lipoproteins in Mammary Gland of Cows

Uchide

Onda

Bonkobara

et al. 1999

The Journal of Veterinary Medical Science

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ABSTRACT. Elution profiles of total lipoproteins, apolipoprotein B (apoB) concentrations in lipoproteins, and plasma triglyceride (TG) levels were examined in early-, late-, and non-lactating cows. Additionally, arteriovenous (A-V) differences were also measured to elucidate the uptake of TG and apoB-containing lipoproteins in mammary gland. Non-lactating cows showed three major peaks corresponding to triglyceride-rich lipoprotein (TRL), low density lipoprotein (LDL), and high density lipoprotein (HDL) fraction, whereas both early-and late-lactating cows revealed two peaks corresponding to TRL and HDL. The peak area of TRL in early-and late-lactating cows were significantly (p<0.05) smaller than that in non-lactating cows. The plasma TG levels and apoB-48 concentrations of TRL in early-and late-lactating cows were also significantly (p<0.01) lower. Furthermore, early lactating cows showed significantly (p<0.05) larger A-V differences in both plasma TG and apoB-48 concentration of TRL than those in late-and non-lactating cows. These results suggested that TG in exogenous (intestinal) TRL was utilized for milk fat synthesis in lactating mammary gland of cows by the receptormediated uptake.-KEY WORDS: apolipoprotein B, cattle, lipoprotein profile, mammary gland, triglyceride-rich lipoprotein.

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Electrophoretic Characterization of Bovine Lipoprotein Subfractions Isolated by Agarose Gel Chromatography

Cited by 20 publications

References 17 publications

Bovine oviductal fluid components and their potential role in sperm cholesterol efflux

Bovine oviductal fluid components and their potential role in sperm cholesterol efflux

Comparison of ultracentrifugation and gel filtration for the isolation of bovine lipoproteins

Utilization of Intestinal Triglyceride-rich Lipoproteins in Mammary Gland of Cows

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