Dual regulation by cAMP of beta-hexosaminidase-induced mitogenesis in bovine tracheal myocytes.

D. B. Lew, C. Nebigil, Kafait Malik

Research output: Contribution to journalArticle

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Abstract

beta-Hexosaminidases, potent mitogens in bovine tracheal myocytes (BTM), stimulate a rapid and transient increase in intracellular cyclic adenosine monophosphate (cAMP) accumulation. The objective of this study was to elucidate the contribution of cAMP in hexosaminidase-induced airway muscle proliferation. Rate of DNA synthesis was measured by 3H-thymidine incorporation in quiescent cells prepared by a low-serum treatment (0.4%) for 48 h after reaching confluency in microtiter wells. cAMP accumulation was measured in acetylated cell extracts in the presence of isobutyl methylxanthine (100 microM) by radioimmunoassay using 125I-cAMP as tracer. Exposure of quiescent cells to purified human placental hexosaminidase B (5 micrograms/ml, 50 nM) caused a significant transient increase in cAMP accumulation (49 to 107 fmol/micrograms protein, or a 20- to 70-fold increase from basal level). Maximum increase occurred at 15 min followed by a rapid decline in cAMP accumulation within 30 min after exposure to hexosaminidase. Similar results were obtained in cells treated with neoglycoprotein mannose bovine serum albumin (100 to 500 nM). The increase in cAMP accumulation was inhibited by mannan (mannose receptor blocker, 0.1 mg/ml), as well as phenylisopropyladenosine (PIA; A1 receptor agonist that inhibits adenylyl cyclase, 0.1 to 1.0 microM). The increase in 3H-thymidine incorporation induced by hexosaminidase B was also inhibited by mannan and PIA. Exposure to 8-(4-chlorophenylthio)-cAMP (cpt-cAMP; a cell-permeable analog of cAMP, 100 microM) or forskolin (a direct activator of catalytic subunit of adenylyl cyclase, 24 microM) up to 6 h enhanced 3H-thymidine incorporation. In contrast, a prolonged exposure (18 to 30 h) to these agents inhibited 3H-thymidine incorporation.(ABSTRACT TRUNCATED AT 250 WORDS)

Original languageEnglish (US)
Pages (from-to)614-619
Number of pages6
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume7
Issue number6
DOIs
StatePublished - Jan 1 1992

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beta-N-Acetylhexosaminidases
Cyclic AMP
Muscle Cells
Thymidine
Hexosaminidase B
Mannans
Hexosaminidases
Adenylyl Cyclases
Phenylisopropyladenosine
Colforsin
Cell Extracts
Mitogens
Radioimmunoassay
Muscle
Catalytic Domain
Glycoproteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Dual regulation by cAMP of beta-hexosaminidase-induced mitogenesis in bovine tracheal myocytes. / Lew, D. B.; Nebigil, C.; Malik, Kafait.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 7, No. 6, 01.01.1992, p. 614-619.

Research output: Contribution to journalArticle

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