Comparison of expression of contractile proteins and myosin heavy chain isoforms (MHCI) in ovine aorta (Ao) and bladder from early fetal through late neonatal periods

Y. H J M Areas, C. R. Rosenfeld, W. Jabbar, K. E. Kamm

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Abstract

Smooth muscle (SM1 and SM2) and nonmuscle (MHC-B) MHCI in Ao and bladder are developmentally regulated. However, their patterns of expression are not fully described, and it is unclear if changes occur with hormonal or functional alterations prior to or at birth. Thus we quantified MHCI and contractile proteins over a prolonged developmental period using Ao and bladder from female fetal (n= 19, 72 to 140d; term- 143d) and newborn (n=25; 1 to 120d) sheep. Tissues were analyzed for total and soluble protein contents (μg/mg wet weight), actin, MHC and MHCf contents (μg/mg wet weight) by SDS-PAOE, using 3-20% gradient and 4% gels, and 200 kDa MHCI by Western Analysis with SM2 and MHC-B specific antisera. Ao proteins gradually rose between ≤ 100d gestation (n=6) and ≥3 mos after birth (n=9): total protein from 65±6 [SEM] to 92±8 (R=0.40, p=0.007), soluble protein 37±3 to 58±5 (R=0.48, p=0.001), actin 3.9±0.9 to 16±2 (R=0.62, p<0.001), and MHC 2.1±0.2 to 4.5±0.5 (R=0.52, p<0.001). Actin/MHC ratios also rose 2.4-fold(R=0.62, p<0.001). In contrast, bladder proteins generally increased (p<0.001, ANOVA) before birth and remained unchanged thereafter: total protein from 46±7 (≤100d gestation, n=6) to 78±4 (130-140d gestation, n=8) to 81±6 (≥3 mos after birth, n=9); soluble protein 38±5 to 58±5 to 69±5, and actin 4.4±0.3 to 12±2 to 19±1; MHC, however, gradually rose from 2.6±0.2 to 6.5±0.6 (R=0.62, p<0.001). 204 kDa MHCI content in Ao and bladder increased 4.3-fold (R=0.68) and 1.5-fold (R=0.58; p<0.001), respectively, by ≥3 mos. Although 200 kDa MHCI content in Ao was unchanged, averaging 1.4±0.1 throughout development, expression of SM2 increased 80% while MHC-B fell proportionally. In contrast, bladder 200 kDa MHCI content rose throughout development, increasing 3-fold by ≥3 mos after birth (R=0.64, p<0.001); however, only SM2 was expressed. Whereas Ao proteins and MHCI gradually rise throughout development, demonstrating no association with known prenatal events, most bladder proteins increase significantly during the last 3 wks of gestation and the only 200 kDa MHCI expressed was SM2. These differences in the patterns of smooth muscle protein expression and ontogeny could be due to differing function of these two smooth muscle types during development.

Original languageEnglish (US)
JournalJournal of Investigative Medicine
Volume44
Issue number1
StatePublished - 1996

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Contractile Proteins
Myosin Heavy Chains
Aorta
Sheep
Protein Isoforms
Urinary Bladder
Proteins
Parturition
Actins
Smooth Muscle
Pregnancy
Muscle
Weights and Measures
Muscle Development
Muscle Proteins
Analysis of variance (ANOVA)
Immune Sera
Analysis of Variance
Gels
Tissue

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

@article{4fb32d2475a44a6683a9eff3756a1a4f,
title = "Comparison of expression of contractile proteins and myosin heavy chain isoforms (MHCI) in ovine aorta (Ao) and bladder from early fetal through late neonatal periods",
abstract = "Smooth muscle (SM1 and SM2) and nonmuscle (MHC-B) MHCI in Ao and bladder are developmentally regulated. However, their patterns of expression are not fully described, and it is unclear if changes occur with hormonal or functional alterations prior to or at birth. Thus we quantified MHCI and contractile proteins over a prolonged developmental period using Ao and bladder from female fetal (n= 19, 72 to 140d; term- 143d) and newborn (n=25; 1 to 120d) sheep. Tissues were analyzed for total and soluble protein contents (μg/mg wet weight), actin, MHC and MHCf contents (μg/mg wet weight) by SDS-PAOE, using 3-20{\%} gradient and 4{\%} gels, and 200 kDa MHCI by Western Analysis with SM2 and MHC-B specific antisera. Ao proteins gradually rose between ≤ 100d gestation (n=6) and ≥3 mos after birth (n=9): total protein from 65±6 [SEM] to 92±8 (R=0.40, p=0.007), soluble protein 37±3 to 58±5 (R=0.48, p=0.001), actin 3.9±0.9 to 16±2 (R=0.62, p<0.001), and MHC 2.1±0.2 to 4.5±0.5 (R=0.52, p<0.001). Actin/MHC ratios also rose 2.4-fold(R=0.62, p<0.001). In contrast, bladder proteins generally increased (p<0.001, ANOVA) before birth and remained unchanged thereafter: total protein from 46±7 (≤100d gestation, n=6) to 78±4 (130-140d gestation, n=8) to 81±6 (≥3 mos after birth, n=9); soluble protein 38±5 to 58±5 to 69±5, and actin 4.4±0.3 to 12±2 to 19±1; MHC, however, gradually rose from 2.6±0.2 to 6.5±0.6 (R=0.62, p<0.001). 204 kDa MHCI content in Ao and bladder increased 4.3-fold (R=0.68) and 1.5-fold (R=0.58; p<0.001), respectively, by ≥3 mos. Although 200 kDa MHCI content in Ao was unchanged, averaging 1.4±0.1 throughout development, expression of SM2 increased 80{\%} while MHC-B fell proportionally. In contrast, bladder 200 kDa MHCI content rose throughout development, increasing 3-fold by ≥3 mos after birth (R=0.64, p<0.001); however, only SM2 was expressed. Whereas Ao proteins and MHCI gradually rise throughout development, demonstrating no association with known prenatal events, most bladder proteins increase significantly during the last 3 wks of gestation and the only 200 kDa MHCI expressed was SM2. These differences in the patterns of smooth muscle protein expression and ontogeny could be due to differing function of these two smooth muscle types during development.",
author = "Areas, {Y. H J M} and Rosenfeld, {C. R.} and W. Jabbar and Kamm, {K. E.}",
year = "1996",
language = "English (US)",
volume = "44",
journal = "Journal of Investigative Medicine",
issn = "1081-5589",
publisher = "Lippincott Williams and Wilkins",
number = "1",

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TY - JOUR

T1 - Comparison of expression of contractile proteins and myosin heavy chain isoforms (MHCI) in ovine aorta (Ao) and bladder from early fetal through late neonatal periods

AU - Areas, Y. H J M

AU - Rosenfeld, C. R.

AU - Jabbar, W.

AU - Kamm, K. E.

PY - 1996

Y1 - 1996

N2 - Smooth muscle (SM1 and SM2) and nonmuscle (MHC-B) MHCI in Ao and bladder are developmentally regulated. However, their patterns of expression are not fully described, and it is unclear if changes occur with hormonal or functional alterations prior to or at birth. Thus we quantified MHCI and contractile proteins over a prolonged developmental period using Ao and bladder from female fetal (n= 19, 72 to 140d; term- 143d) and newborn (n=25; 1 to 120d) sheep. Tissues were analyzed for total and soluble protein contents (μg/mg wet weight), actin, MHC and MHCf contents (μg/mg wet weight) by SDS-PAOE, using 3-20% gradient and 4% gels, and 200 kDa MHCI by Western Analysis with SM2 and MHC-B specific antisera. Ao proteins gradually rose between ≤ 100d gestation (n=6) and ≥3 mos after birth (n=9): total protein from 65±6 [SEM] to 92±8 (R=0.40, p=0.007), soluble protein 37±3 to 58±5 (R=0.48, p=0.001), actin 3.9±0.9 to 16±2 (R=0.62, p<0.001), and MHC 2.1±0.2 to 4.5±0.5 (R=0.52, p<0.001). Actin/MHC ratios also rose 2.4-fold(R=0.62, p<0.001). In contrast, bladder proteins generally increased (p<0.001, ANOVA) before birth and remained unchanged thereafter: total protein from 46±7 (≤100d gestation, n=6) to 78±4 (130-140d gestation, n=8) to 81±6 (≥3 mos after birth, n=9); soluble protein 38±5 to 58±5 to 69±5, and actin 4.4±0.3 to 12±2 to 19±1; MHC, however, gradually rose from 2.6±0.2 to 6.5±0.6 (R=0.62, p<0.001). 204 kDa MHCI content in Ao and bladder increased 4.3-fold (R=0.68) and 1.5-fold (R=0.58; p<0.001), respectively, by ≥3 mos. Although 200 kDa MHCI content in Ao was unchanged, averaging 1.4±0.1 throughout development, expression of SM2 increased 80% while MHC-B fell proportionally. In contrast, bladder 200 kDa MHCI content rose throughout development, increasing 3-fold by ≥3 mos after birth (R=0.64, p<0.001); however, only SM2 was expressed. Whereas Ao proteins and MHCI gradually rise throughout development, demonstrating no association with known prenatal events, most bladder proteins increase significantly during the last 3 wks of gestation and the only 200 kDa MHCI expressed was SM2. These differences in the patterns of smooth muscle protein expression and ontogeny could be due to differing function of these two smooth muscle types during development.

AB - Smooth muscle (SM1 and SM2) and nonmuscle (MHC-B) MHCI in Ao and bladder are developmentally regulated. However, their patterns of expression are not fully described, and it is unclear if changes occur with hormonal or functional alterations prior to or at birth. Thus we quantified MHCI and contractile proteins over a prolonged developmental period using Ao and bladder from female fetal (n= 19, 72 to 140d; term- 143d) and newborn (n=25; 1 to 120d) sheep. Tissues were analyzed for total and soluble protein contents (μg/mg wet weight), actin, MHC and MHCf contents (μg/mg wet weight) by SDS-PAOE, using 3-20% gradient and 4% gels, and 200 kDa MHCI by Western Analysis with SM2 and MHC-B specific antisera. Ao proteins gradually rose between ≤ 100d gestation (n=6) and ≥3 mos after birth (n=9): total protein from 65±6 [SEM] to 92±8 (R=0.40, p=0.007), soluble protein 37±3 to 58±5 (R=0.48, p=0.001), actin 3.9±0.9 to 16±2 (R=0.62, p<0.001), and MHC 2.1±0.2 to 4.5±0.5 (R=0.52, p<0.001). Actin/MHC ratios also rose 2.4-fold(R=0.62, p<0.001). In contrast, bladder proteins generally increased (p<0.001, ANOVA) before birth and remained unchanged thereafter: total protein from 46±7 (≤100d gestation, n=6) to 78±4 (130-140d gestation, n=8) to 81±6 (≥3 mos after birth, n=9); soluble protein 38±5 to 58±5 to 69±5, and actin 4.4±0.3 to 12±2 to 19±1; MHC, however, gradually rose from 2.6±0.2 to 6.5±0.6 (R=0.62, p<0.001). 204 kDa MHCI content in Ao and bladder increased 4.3-fold (R=0.68) and 1.5-fold (R=0.58; p<0.001), respectively, by ≥3 mos. Although 200 kDa MHCI content in Ao was unchanged, averaging 1.4±0.1 throughout development, expression of SM2 increased 80% while MHC-B fell proportionally. In contrast, bladder 200 kDa MHCI content rose throughout development, increasing 3-fold by ≥3 mos after birth (R=0.64, p<0.001); however, only SM2 was expressed. Whereas Ao proteins and MHCI gradually rise throughout development, demonstrating no association with known prenatal events, most bladder proteins increase significantly during the last 3 wks of gestation and the only 200 kDa MHCI expressed was SM2. These differences in the patterns of smooth muscle protein expression and ontogeny could be due to differing function of these two smooth muscle types during development.

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