In vitro studies on adult cardiac myocytes

Attachment and biosynthesis of collagen type IV and laminin

E. Lundgren, D. Gullberg, K. Rubin, T. K. Borg, M. J. Terracio, Louis Terracio

Research output: Contribution to journalArticle

Abstract

The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when alminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.

Original languageEnglish (US)
Pages (from-to)43-53
Number of pages11
JournalJournal of Cellular Physiology
Volume136
Issue number1
StatePublished - 1988

Fingerprint

Collagen Type IV
Biosynthesis
Laminin
Cardiac Myocytes
Muscle Cells
Staining and Labeling
Fluorescent Antibody Technique
In Vitro Techniques
Pseudopodia
Antibody Affinity
Antibodies
Pepsin A
Collagen Type I
Immunoprecipitation
Basement Membrane
Labeling
Rats
Collagen

ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Physiology

Cite this

In vitro studies on adult cardiac myocytes : Attachment and biosynthesis of collagen type IV and laminin. / Lundgren, E.; Gullberg, D.; Rubin, K.; Borg, T. K.; Terracio, M. J.; Terracio, Louis.

In: Journal of Cellular Physiology, Vol. 136, No. 1, 1988, p. 43-53.

Research output: Contribution to journalArticle

Lundgren, E, Gullberg, D, Rubin, K, Borg, TK, Terracio, MJ & Terracio, L 1988, 'In vitro studies on adult cardiac myocytes: Attachment and biosynthesis of collagen type IV and laminin', Journal of Cellular Physiology, vol. 136, no. 1, pp. 43-53.
Lundgren, E. ; Gullberg, D. ; Rubin, K. ; Borg, T. K. ; Terracio, M. J. ; Terracio, Louis. / In vitro studies on adult cardiac myocytes : Attachment and biosynthesis of collagen type IV and laminin. In: Journal of Cellular Physiology. 1988 ; Vol. 136, No. 1. pp. 43-53.
@article{50cf9c98b17841d2b81eff61d6f56d0e,
title = "In vitro studies on adult cardiac myocytes: Attachment and biosynthesis of collagen type IV and laminin",
abstract = "The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when alminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.",
author = "E. Lundgren and D. Gullberg and K. Rubin and Borg, {T. K.} and Terracio, {M. J.} and Louis Terracio",
year = "1988",
language = "English (US)",
volume = "136",
pages = "43--53",
journal = "Journal of Cellular Physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "1",

}

TY - JOUR

T1 - In vitro studies on adult cardiac myocytes

T2 - Attachment and biosynthesis of collagen type IV and laminin

AU - Lundgren, E.

AU - Gullberg, D.

AU - Rubin, K.

AU - Borg, T. K.

AU - Terracio, M. J.

AU - Terracio, Louis

PY - 1988

Y1 - 1988

N2 - The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when alminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.

AB - The interactions between adult rat cardiac myocytes and the basement membrane components collagen type IV and laminin were investigated in attachment experiments and biosynthesis studies and by immunofluorescence staining. Adult myocytes attached equally well to native collagen type IV and laminin but did not attach to collagen type IV solubilized with pepsin (P-CIV) or to collagen type I. However, when alminin was used to coat P-CIV, attachment was enhanced. Affinity-purified antibodies against laminin inhibited the attachment of myocytes to dishes coated with native collagen type IV, indicating that cell surface-bound laminin mediated attachment of the cells to this substrate. Immunofluorescence staining of freshly isolated myocytes, using antibodies against laminin or collagen type IV, revealed the presence of laminin but not of collagen type IV on the surface of freshly isolated cells, indicating that during the isolation procedure collagen IV was removed from the cell surface. Metabolic labeling followed by immunoprecipitation demonstrated synthesis of both laminin and collagen type IV in cardiac myocytes as they progressed into culture over a 14-day period. This synthesis was accompanied by the deposition of the collagen type IV and laminin into distinctly different patterns as revealed by immunofluorescence staining. As the cells progressed into culture, newly synthesized laminin formed a network radiating from the center of the reorganizing cell into the pseudopods. The laminin was redistributed and remodeled with time in culture to form a dense layer beneath the cell. Collagen type IV was also synthesized with time in culture, but the pattern was a much finer network as opposed to the denser pattern of laminin staining. These studies demonstrate that adult cardiac myocytes synthesize and remodel the basement membrane as they adapt to the culture environment.

UR - http://www.scopus.com/inward/record.url?scp=0023678642&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023678642&partnerID=8YFLogxK

M3 - Article

VL - 136

SP - 43

EP - 53

JO - Journal of Cellular Physiology

JF - Journal of Cellular Physiology

SN - 0021-9541

IS - 1

ER -