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. 1988 Oct 1;107(4):1317–1323. doi: 10.1083/jcb.107.4.1317

Expression of a testis-specific hsp70 gene-related RNA in defined stages of rat seminiferous epithelium

PMCID: PMC2115240  PMID: 3170632

Abstract

Changes in the level of a testis-specific hsp70 gene-related transcript (hst70 RNA) and its cellular localization during the cycle of rat seminiferous epithelium have been investigated. Segments of seminiferous tubules at defined stages of the cycle were isolated in living condition by transillumination-assisted microdissection and the exact stages identified by phase-contrast microscopy of live cell squashes. The levels of the hst70 RNA were determined by Northern and slot blotting of whole cell lysates. High levels were found in stages XII-XIV and I to early VII of the cycle, and low levels were found in other stages, i.e., late VII (VIId) through VIII-XI of the cycle. The in situ hybridization revealed that the hst70 gene was activated in late pachytene primary spermatocytes during stage XII of the cycle, and that mRNA was then present in cells during differentiation through diakinesis, meiotic divisions, and early spermiogenesis (steps 1 through early 7). The activation of the gene coding for hst70 RNA shortly before meiotic divisions may indicate that the gene product is needed either during differentiation of late spermatocytes into spermatids or later during spermiogenesis, and that the mRNA may be stored in early spermatids.

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Selected References

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  1. Allen R. L., O'Brien D. A., Eddy E. M. A novel hsp70-like protein (P70) is present in mouse spermatogenic cells. Mol Cell Biol. 1988 Feb;8(2):828–832. doi: 10.1128/mcb.8.2.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arnheim N. Characterization of mouse ribosomal gene fragments purified by molecular cloning. Gene. 1979 Oct;7(2):83–96. doi: 10.1016/0378-1119(79)90025-8. [DOI] [PubMed] [Google Scholar]
  3. Bardwell J. C., Craig E. A. Major heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous. Proc Natl Acad Sci U S A. 1984 Feb;81(3):848–852. doi: 10.1073/pnas.81.3.848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bensaude O., Babinet C., Morange M., Jacob F. Heat shock proteins, first major products of zygotic gene activity in mouse embryo. Nature. 1983 Sep 22;305(5932):331–333. doi: 10.1038/305331a0. [DOI] [PubMed] [Google Scholar]
  5. Brahic M., Haase A. T. Detection of viral sequences of low reiteration frequency by in situ hybridization. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6125–6129. doi: 10.1073/pnas.75.12.6125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carper S. W., Duffy J. J., Gerner E. W. Heat shock proteins in thermotolerance and other cellular processes. Cancer Res. 1987 Oct 15;47(20):5249–5255. [PubMed] [Google Scholar]
  7. Carr B. I., Huang T. H., Buzin C. H., Itakura K. Induction of heat shock gene expression without heat shock by hepatocarcinogens and during hepatic regeneration in rat liver. Cancer Res. 1986 Oct;46(10):5106–5111. [PubMed] [Google Scholar]
  8. Craig E. A., Ingolia T. D., Manseau L. J. Expression of Drosophila heat-shock cognate genes during heat shock and development. Dev Biol. 1983 Oct;99(2):418–426. doi: 10.1016/0012-1606(83)90291-9. [DOI] [PubMed] [Google Scholar]
  9. Craig E. A. The heat shock response. CRC Crit Rev Biochem. 1985;18(3):239–280. doi: 10.3109/10409238509085135. [DOI] [PubMed] [Google Scholar]
  10. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  11. Gizang-Ginsberg E., Wolgemuth D. J. Expression of the proopiomelanocortin gene is developmentally regulated and affected by germ cells in the male mouse reproductive system. Proc Natl Acad Sci U S A. 1987 Mar;84(6):1600–1604. doi: 10.1073/pnas.84.6.1600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goldman D. S., Kiessling A. A., Millette C. F., Cooper G. M. Expression of c-mos RNA in germ cells of male and female mice. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4509–4513. doi: 10.1073/pnas.84.13.4509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hahnel A. C., Gifford D. J., Heikkila J. J., Schultz G. A. Expression of the major heat shock protein (hsp 70) family during early mouse embryo development. Teratog Carcinog Mutagen. 1986;6(6):493–510. doi: 10.1002/tcm.1770060603. [DOI] [PubMed] [Google Scholar]
  14. Hecht N. B., Bower P. A., Waters S. H., Yelick P. C., Distel R. J. Evidence for haploid expression of mouse testicular genes. Exp Cell Res. 1986 May;164(1):183–190. doi: 10.1016/0014-4827(86)90465-9. [DOI] [PubMed] [Google Scholar]
  15. Heidaran M. A., Kistler W. S. Transcriptional and translational control of the message for transition protein 1, a major chromosomal protein of mammalian spermatids. J Biol Chem. 1987 Sep 25;262(27):13309–13315. [PubMed] [Google Scholar]
  16. Heikkila J. J., Browder L. W., Gedamu L., Nickells R. W., Schultz G. A. Heat-shock gene expression in animal embryonic systems. Can J Genet Cytol. 1986 Dec;28(6):1093–1105. doi: 10.1139/g86-153. [DOI] [PubMed] [Google Scholar]
  17. Hunt C., Morimoto R. I. Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6455–6459. doi: 10.1073/pnas.82.19.6455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ingolia T. D., Slater M. R., Craig E. A. Saccharomyces cerevisiae contains a complex multigene family related to the major heat shock-inducible gene of Drosophila. Mol Cell Biol. 1982 Nov;2(11):1388–1398. doi: 10.1128/mcb.2.11.1388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kaczmarek L., Calabretta B., Kao H. T., Heintz N., Nevins J., Baserga R. Control of hsp70 RNA levels in human lymphocytes. J Cell Biol. 1987 Feb;104(2):183–187. doi: 10.1083/jcb.104.2.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kao H. T., Capasso O., Heintz N., Nevins J. R. Cell cycle control of the human HSP70 gene: implications for the role of a cellular E1A-like function. Mol Cell Biol. 1985 Apr;5(4):628–633. doi: 10.1128/mcb.5.4.628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kilpatrick D. L., Millette C. F. Expression of proenkephalin messenger RNA by mouse spermatogenic cells. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5015–5018. doi: 10.1073/pnas.83.14.5015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kim Y. J., Hwang I., Tres L. L., Kierszenbaum A. L., Chae C. B. Molecular cloning and differential expression of somatic and testis-specific H2B histone genes during rat spermatogenesis. Dev Biol. 1987 Nov;124(1):23–34. doi: 10.1016/0012-1606(87)90455-6. [DOI] [PubMed] [Google Scholar]
  23. Krawczyk Z., Szymik N., Wiśniewski J. Expression of hsp70-related gene in developing and degenerating rat testis. Mol Biol Rep. 1987;12(1):35–41. doi: 10.1007/BF00580648. [DOI] [PubMed] [Google Scholar]
  24. Krawczyk Z., Wiśniewski J., Biesiada E. A hsp70-related gene is constitutively highly expressed in testis of rat and mouse. Mol Biol Rep. 1987;12(1):27–34. doi: 10.1007/BF00580647. [DOI] [PubMed] [Google Scholar]
  25. Krawczyk Z., Wu C. Isolation of RNA for dot hybridization by heparin-DNase I treatment of whole cell lysate. Anal Biochem. 1987 Aug 15;165(1):20–27. doi: 10.1016/0003-2697(87)90195-3. [DOI] [PubMed] [Google Scholar]
  26. LEBLOND C. P., CLERMONT Y. Definition of the stages of the cycle of the seminiferous epithelium in the rat. Ann N Y Acad Sci. 1952 Nov 20;55(4):548–573. doi: 10.1111/j.1749-6632.1952.tb26576.x. [DOI] [PubMed] [Google Scholar]
  27. McCarrey J. R., Thomas K. Human testis-specific PGK gene lacks introns and possesses characteristics of a processed gene. Nature. 1987 Apr 2;326(6112):501–505. doi: 10.1038/326501a0. [DOI] [PubMed] [Google Scholar]
  28. Milarski K. L., Morimoto R. I. Expression of human HSP70 during the synthetic phase of the cell cycle. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9517–9521. doi: 10.1073/pnas.83.24.9517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Millan J. L., Driscoll C. E., LeVan K. M., Goldberg E. Epitopes of human testis-specific lactate dehydrogenase deduced from a cDNA sequence. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5311–5315. doi: 10.1073/pnas.84.15.5311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mues G. I., Munn T. Z., Raese J. D. A human gene family with sequence homology to Drosophila melanogaster Hsp70 heat shock genes. J Biol Chem. 1986 Jan 15;261(2):874–877. [PubMed] [Google Scholar]
  31. Munro S., Pelham H. R. An Hsp70-like protein in the ER: identity with the 78 kd glucose-regulated protein and immunoglobulin heavy chain binding protein. Cell. 1986 Jul 18;46(2):291–300. doi: 10.1016/0092-8674(86)90746-4. [DOI] [PubMed] [Google Scholar]
  32. Mutter G. L., Wolgemuth D. J. Distinct developmental patterns of c-mos protooncogene expression in female and male mouse germ cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5301–5305. doi: 10.1073/pnas.84.15.5301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. O'Malley K., Mauron A., Barchas J. D., Kedes L. Constitutively expressed rat mRNA encoding a 70-kilodalton heat-shock-like protein. Mol Cell Biol. 1985 Dec;5(12):3476–3483. doi: 10.1128/mcb.5.12.3476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Parvinen M., Vanha-Perttula T. Identification and enzyme quantitation of the stages of the seminiferous epithelial wave in the rat. Anat Rec. 1972 Dec;174(4):435–449. doi: 10.1002/ar.1091740404. [DOI] [PubMed] [Google Scholar]
  35. Pelham H. R. Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell. 1986 Sep 26;46(7):959–961. doi: 10.1016/0092-8674(86)90693-8. [DOI] [PubMed] [Google Scholar]
  36. Peschon J. J., Behringer R. R., Brinster R. L., Palmiter R. D. Spermatid-specific expression of protamine 1 in transgenic mice. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5316–5319. doi: 10.1073/pnas.84.15.5316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ponzetto C., Wolgemuth D. J. Haploid expression of a unique c-abl transcript in the mouse male germ line. Mol Cell Biol. 1985 Jul;5(7):1791–1794. doi: 10.1128/mcb.5.7.1791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rubin M. R., Toth L. E., Patel M. D., D'Eustachio P., Nguyen-Huu M. C. A mouse homeo box gene is expressed in spermatocytes and embryos. Science. 1986 Aug 8;233(4764):663–667. doi: 10.1126/science.3726554. [DOI] [PubMed] [Google Scholar]
  39. Sandberg M., Vuorio E. Localization of types I, II, and III collagen mRNAs in developing human skeletal tissues by in situ hybridization. J Cell Biol. 1987 Apr;104(4):1077–1084. doi: 10.1083/jcb.104.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shackleford G. M., Varmus H. E. Expression of the proto-oncogene int-1 is restricted to postmeiotic male germ cells and the neural tube of mid-gestational embryos. Cell. 1987 Jul 3;50(1):89–95. doi: 10.1016/0092-8674(87)90665-9. [DOI] [PubMed] [Google Scholar]
  41. Sorger P. K., Pelham H. R. Cloning and expression of a gene encoding hsc73, the major hsp70-like protein in unstressed rat cells. EMBO J. 1987 Apr;6(4):993–998. doi: 10.1002/j.1460-2075.1987.tb04850.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stewart T. A., Bellvé A. R., Leder P. Transcription and promoter usage of the myc gene in normal somatic and spermatogenic cells. Science. 1984 Nov 9;226(4675):707–710. doi: 10.1126/science.6494906. [DOI] [PubMed] [Google Scholar]
  43. Sullivan K. F., Machlin P. S., Ratrie H., 3rd, Cleveland D. W. Sequence and expression of the chicken beta 3 tubulin gene. A vertebrate testis beta-tubulin isotype. J Biol Chem. 1986 Oct 5;261(28):13317–13322. [PubMed] [Google Scholar]
  44. Syrjänen S., Syrjänen K., Mäntyjärvi R., Parkkinen S., Väyrynen M., Saarikoski S., Castren O. Human papillomavirus (HPV) DNA sequences demonstrated by in situ DNA hybridization in serial paraffin-embedded cervical biopsies. Arch Gynecol. 1986;239(1):39–48. doi: 10.1007/BF02134287. [DOI] [PubMed] [Google Scholar]
  45. Söderström K. O., Parvinen M. RNA synthesis in different stages of rat seminiferous epithelial cycle. Mol Cell Endocrinol. 1976 Aug-Sep;5(3-4):181–199. doi: 10.1016/0303-7207(76)90082-4. [DOI] [PubMed] [Google Scholar]
  46. Tanaka S., Fujimoto H. A postmeiotically expressed clone encodes lactate dehydrogenase isozyme X. Biochem Biophys Res Commun. 1986 Apr 29;136(2):760–766. doi: 10.1016/0006-291x(86)90504-8. [DOI] [PubMed] [Google Scholar]
  47. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Toppari J., Parvinen M. In vitro differentiation of rat seminiferous tubular segments from defined stages of the epithelial cycle morphologic and immunolocalization analysis. J Androl. 1985 Nov-Dec;6(6):334–343. doi: 10.1002/j.1939-4640.1985.tb03289.x. [DOI] [PubMed] [Google Scholar]
  49. Villasante A., Wang D., Dobner P., Dolph P., Lewis S. A., Cowan N. J. Six mouse alpha-tubulin mRNAs encode five distinct isotypes: testis-specific expression of two sister genes. Mol Cell Biol. 1986 Jul;6(7):2409–2419. doi: 10.1128/mcb.6.7.2409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Voellmy R., Ahmed A., Schiller P., Bromley P., Rungger D. Isolation and functional analysis of a human 70,000-dalton heat shock protein gene segment. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4949–4953. doi: 10.1073/pnas.82.15.4949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Waters S. H., Distel R. J., Hecht N. B. Mouse testes contain two size classes of actin mRNA that are differentially expressed during spermatogenesis. Mol Cell Biol. 1985 Jul;5(7):1649–1654. doi: 10.1128/mcb.5.7.1649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wing T. Y., Christensen A. K. Morphometric studies on rat seminiferous tubules. Am J Anat. 1982 Sep;165(1):13–25. doi: 10.1002/aja.1001650103. [DOI] [PubMed] [Google Scholar]
  53. Wolgemuth D. J., Gizang-Ginsberg E., Engelmyer E., Gavin B. J., Ponzetto C. Separation of mouse testis cells on a Celsep (TM) apparatus and their usefulness as a source of high molecular weight DNA or RNA. Gamete Res. 1985;12:1–10. doi: 10.1002/mrd.1120120102. [DOI] [PubMed] [Google Scholar]
  54. Wolgemuth D. J., Viviano C. M., Gizang-Ginsberg E., Frohman M. A., Joyner A. L., Martin G. R. Differential expression of the mouse homeobox-containing gene Hox-1.4 during male germ cell differentiation and embryonic development. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5813–5817. doi: 10.1073/pnas.84.16.5813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zakeri Z. F., Wolgemuth D. J. Developmental-stage-specific expression of the hsp70 gene family during differentiation of the mammalian male germ line. Mol Cell Biol. 1987 May;7(5):1791–1796. doi: 10.1128/mcb.7.5.1791. [DOI] [PMC free article] [PubMed] [Google Scholar]

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