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HP1 knockdown is associated with abnormal condensation of almost all chromatin types in a grasshopper (Eyprepocnemis plorans)

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Abstract

Heterochromatin protein 1 (HP1) is a highly conserved family of eukaryotic proteins required for heterochromatic gene silencing and euchromatic gene transcription regulation. In addition, HP1 is involved in chromatin organization and protection of chromosome integrity during cell division. Here, we present a cytological and molecular analysis of the effects of HP1 knockdown in Eyprepocnemis plorans, a grasshopper species polymorphic for supernumerary heterochromatic chromosomes. Our results revealed contrasting effects of HP1 knockdown on gene activity. While the Bub1 gene decreased in expression level in HP1 knockdown animals, NOR activity, rRNA and, contrarily to previous reports in Drosophila, Hsp70 gene expression remained unchanged. Furthermore, HP1 knockdown resulted in abnormal chromatin condensation, chromosomal bridges, higher frequency of macrospermatids, loss of muscle mass and hemolymph amount as well as a low number of dividing cells and survival reduction. All these phenotypes are very likely due to the chromatin condensation disruption observed for almost all kinds of chromatin.

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Abbreviations

Bub1:

Budding uninhibited by benzimidazoles 1

cDNA:

Complementary deoxyribonucleic acid

Dffi:

Days from the first injection

dsRNA:

Double-stranded ribosomal ribonucleic acid

E. plorans :

Eyprepocnemis plorans

FISH:

Fluorescent in situ hibridization

gDNA:

Genomic deoxyribonucleic acid

HKG:

Housekeeping gene

HP1:

Heterochromatin protein 1

Hsp70:

70 kilodaltons heat shock protein

ITS2:

Internal transcribed spacer 2

NOR:

Nucleolus organizer region

qRT-PCR:

Quantitative reverse transcription-polymerase chain reaction

rDNA:

Ribosomal deoxyribonucleic acid

rRNA:

Ribosomal ribonucleic acid

RNAi:

Interference ribosomal ribonucleic acid

SAC:

Spindle assembly checkpoint

satDNA:

Satellite deoxyribonucleic acid

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Acknowledgements

We thank T. López for technical assistance and Karl Meunier for language revision. This study was supported by a grant from the Spanish Ministerio de Ciencia e Innovación (CGL2009-11917) and Plan Andaluz de Investigación (CVI-6649), and was partially performed by FEDER funds. M Ruiz-Estévez was supported by a FPU fellowship from the Spanish Ministerio de Ciencia e Innovación. M. Bakkali wishes to thank the Spanish Ministerio de Ciencia e Innovación both for the grant (BFU2010-16438) and for the Ramón y Cajal Fellowship.

Integrity of research

All experiments comply with the current Spanish laws. All institutional and national guidelines for the care and use of laboratory animals were followed.

Conflict of interest

Mercedes Ruíz-Estévez, Mohammed Bakkali, Josefa Cabrero, Juan Pedro M Camacho and María Dolores López-León declare that they have no conflict of interest.

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  1. Departamento de Genética, Facultad de Ciencias, Universidad de Granada, 18071, Granada, Spain

    Mercedes Ruiz-Estévez, Mohammed Bakkali, Josefa Cabrero, Juan Pedro M. Camacho & María Dolores López-León

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  1. Mercedes Ruiz-Estévez
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  2. Mohammed Bakkali
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  3. Josefa Cabrero
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  4. Juan Pedro M. Camacho
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Correspondence to María Dolores López-León.

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Ruiz-Estévez, M., Bakkali, M., Cabrero, J. et al. HP1 knockdown is associated with abnormal condensation of almost all chromatin types in a grasshopper (Eyprepocnemis plorans). Chromosome Res 22, 253–266 (2014). https://doi.org/10.1007/s10577-013-9399-z

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