Stimulation of Tubulin Synthesis by Lactate in Isolated Spermatogenic Cells: tubulin/spermatocytes/spermatids/rat

Masahisa Nakamura, Masayuki Komukai, Ryoichi Matsuda, Shoichi Okinaga, Kiyoshi Arai

Research output: Contribution to journalArticle

Abstract

The effect of lactate on synthesis of new proteins in isolated spermatids and spermatocytes of rats was examined. Lactate stimulated[35S]methionine ([35S]met) incorporation into both spermatids and spermatocytes. The rate of protein synthesis was positively correlated with the intracellular level of ATP. The [35S]met‐labeled proteins in the two types of cells were compared by one and two dimensional polyacrylamide gel electrophoresis (1D and 2D‐PAGE) and autoradiography. The syntheses of several stagespecific and non‐specific proteins were observed. When spermatids and spermatocytes were cultured in medium without lactate, two major proteins of molecular weight (Mr) 43 kD and 55 kD were detected in the water‐soluble fraction (105,000 g supernatant), and one major protein of Mr 24 kD was observed in the membrane‐rich fraction. Addition of lactate to the incubation medium dramatically increased the synthesis of six proteins (Mr 14 kD, 16 kD, 43 kD, 55 kD, 84 kD and 135 kD) in the water‐soluble fractions of spermatids and spermatocytes, but did not stimulate the synthesis of the Mr 24 kD protein in the membrane‐rich fraction. In addition, after 1D and 2D‐PAGE and electrophoretic transfer to nitrocellulose, two proteins of Mr 43 kD and 55 kD were identified as actin and tubulin, respectively, on the basis of their reactivities with specific antisera. Tubulin was also produced by in vitro translation using a spermatid lysate. These results suggest that lactate may play an important role in changing the cell structure and shape during spermatogenesis by regulating the syntheses of actin and tubulin.

Original languageEnglish
Pages (from-to)179-188
Number of pages10
JournalDevelopment Growth and Differentiation
Volume31
Issue number2
DOIs
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

Spermatocytes
Spermatids
Tubulin
Lactic Acid
Proteins
Actins
Collodion
Cell Shape
Electrophoresis, Gel, Two-Dimensional
Spermatogenesis
Autoradiography
Methionine
Immune Sera
Adenosine Triphosphate
Molecular Weight

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Stimulation of Tubulin Synthesis by Lactate in Isolated Spermatogenic Cells : tubulin/spermatocytes/spermatids/rat. / Nakamura, Masahisa; Komukai, Masayuki; Matsuda, Ryoichi; Okinaga, Shoichi; Arai, Kiyoshi.

In: Development Growth and Differentiation, Vol. 31, No. 2, 1989, p. 179-188.

Research output: Contribution to journalArticle

Nakamura, Masahisa ; Komukai, Masayuki ; Matsuda, Ryoichi ; Okinaga, Shoichi ; Arai, Kiyoshi. / Stimulation of Tubulin Synthesis by Lactate in Isolated Spermatogenic Cells : tubulin/spermatocytes/spermatids/rat. In: Development Growth and Differentiation. 1989 ; Vol. 31, No. 2. pp. 179-188.
@article{64ec8b170922444a89578a9d7559adb1,
title = "Stimulation of Tubulin Synthesis by Lactate in Isolated Spermatogenic Cells: tubulin/spermatocytes/spermatids/rat",
abstract = "The effect of lactate on synthesis of new proteins in isolated spermatids and spermatocytes of rats was examined. Lactate stimulated[35S]methionine ([35S]met) incorporation into both spermatids and spermatocytes. The rate of protein synthesis was positively correlated with the intracellular level of ATP. The [35S]met‐labeled proteins in the two types of cells were compared by one and two dimensional polyacrylamide gel electrophoresis (1D and 2D‐PAGE) and autoradiography. The syntheses of several stagespecific and non‐specific proteins were observed. When spermatids and spermatocytes were cultured in medium without lactate, two major proteins of molecular weight (Mr) 43 kD and 55 kD were detected in the water‐soluble fraction (105,000 g supernatant), and one major protein of Mr 24 kD was observed in the membrane‐rich fraction. Addition of lactate to the incubation medium dramatically increased the synthesis of six proteins (Mr 14 kD, 16 kD, 43 kD, 55 kD, 84 kD and 135 kD) in the water‐soluble fractions of spermatids and spermatocytes, but did not stimulate the synthesis of the Mr 24 kD protein in the membrane‐rich fraction. In addition, after 1D and 2D‐PAGE and electrophoretic transfer to nitrocellulose, two proteins of Mr 43 kD and 55 kD were identified as actin and tubulin, respectively, on the basis of their reactivities with specific antisera. Tubulin was also produced by in vitro translation using a spermatid lysate. These results suggest that lactate may play an important role in changing the cell structure and shape during spermatogenesis by regulating the syntheses of actin and tubulin.",
author = "Masahisa Nakamura and Masayuki Komukai and Ryoichi Matsuda and Shoichi Okinaga and Kiyoshi Arai",
year = "1989",
doi = "10.1111/j.1440-169X.1989.00179.x",
language = "English",
volume = "31",
pages = "179--188",
journal = "Development Growth and Differentiation",
issn = "0012-1592",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Stimulation of Tubulin Synthesis by Lactate in Isolated Spermatogenic Cells

T2 - tubulin/spermatocytes/spermatids/rat

AU - Nakamura, Masahisa

AU - Komukai, Masayuki

AU - Matsuda, Ryoichi

AU - Okinaga, Shoichi

AU - Arai, Kiyoshi

PY - 1989

Y1 - 1989

N2 - The effect of lactate on synthesis of new proteins in isolated spermatids and spermatocytes of rats was examined. Lactate stimulated[35S]methionine ([35S]met) incorporation into both spermatids and spermatocytes. The rate of protein synthesis was positively correlated with the intracellular level of ATP. The [35S]met‐labeled proteins in the two types of cells were compared by one and two dimensional polyacrylamide gel electrophoresis (1D and 2D‐PAGE) and autoradiography. The syntheses of several stagespecific and non‐specific proteins were observed. When spermatids and spermatocytes were cultured in medium without lactate, two major proteins of molecular weight (Mr) 43 kD and 55 kD were detected in the water‐soluble fraction (105,000 g supernatant), and one major protein of Mr 24 kD was observed in the membrane‐rich fraction. Addition of lactate to the incubation medium dramatically increased the synthesis of six proteins (Mr 14 kD, 16 kD, 43 kD, 55 kD, 84 kD and 135 kD) in the water‐soluble fractions of spermatids and spermatocytes, but did not stimulate the synthesis of the Mr 24 kD protein in the membrane‐rich fraction. In addition, after 1D and 2D‐PAGE and electrophoretic transfer to nitrocellulose, two proteins of Mr 43 kD and 55 kD were identified as actin and tubulin, respectively, on the basis of their reactivities with specific antisera. Tubulin was also produced by in vitro translation using a spermatid lysate. These results suggest that lactate may play an important role in changing the cell structure and shape during spermatogenesis by regulating the syntheses of actin and tubulin.

AB - The effect of lactate on synthesis of new proteins in isolated spermatids and spermatocytes of rats was examined. Lactate stimulated[35S]methionine ([35S]met) incorporation into both spermatids and spermatocytes. The rate of protein synthesis was positively correlated with the intracellular level of ATP. The [35S]met‐labeled proteins in the two types of cells were compared by one and two dimensional polyacrylamide gel electrophoresis (1D and 2D‐PAGE) and autoradiography. The syntheses of several stagespecific and non‐specific proteins were observed. When spermatids and spermatocytes were cultured in medium without lactate, two major proteins of molecular weight (Mr) 43 kD and 55 kD were detected in the water‐soluble fraction (105,000 g supernatant), and one major protein of Mr 24 kD was observed in the membrane‐rich fraction. Addition of lactate to the incubation medium dramatically increased the synthesis of six proteins (Mr 14 kD, 16 kD, 43 kD, 55 kD, 84 kD and 135 kD) in the water‐soluble fractions of spermatids and spermatocytes, but did not stimulate the synthesis of the Mr 24 kD protein in the membrane‐rich fraction. In addition, after 1D and 2D‐PAGE and electrophoretic transfer to nitrocellulose, two proteins of Mr 43 kD and 55 kD were identified as actin and tubulin, respectively, on the basis of their reactivities with specific antisera. Tubulin was also produced by in vitro translation using a spermatid lysate. These results suggest that lactate may play an important role in changing the cell structure and shape during spermatogenesis by regulating the syntheses of actin and tubulin.

UR - http://www.scopus.com/inward/record.url?scp=84985793226&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84985793226&partnerID=8YFLogxK

U2 - 10.1111/j.1440-169X.1989.00179.x

DO - 10.1111/j.1440-169X.1989.00179.x

M3 - Article

AN - SCOPUS:84985793226

VL - 31

SP - 179

EP - 188

JO - Development Growth and Differentiation

JF - Development Growth and Differentiation

SN - 0012-1592

IS - 2

ER -