Highly purified spermatozoal RNA obtained by a novel method indicates an unusual 28S/18S rRNA ratio and suggests impaired ribosome assembly.
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Highly purified spermatozoal RNA obtained by a novel method indicates an unusual 28S/18S rRNA ratio and suggests impaired ribosome assembly. / Capallo-Obermann, Heike; Schulze, Wolfgang; Jastrow, Holger; Baukloh, Vera; Spiess, Andrej-Nikolai.
In: MOL HUM REPROD, Vol. 17, No. 11, 11, 2011, p. 669-678.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Highly purified spermatozoal RNA obtained by a novel method indicates an unusual 28S/18S rRNA ratio and suggests impaired ribosome assembly.
AU - Capallo-Obermann, Heike
AU - Schulze, Wolfgang
AU - Jastrow, Holger
AU - Baukloh, Vera
AU - Spiess, Andrej-Nikolai
PY - 2011
Y1 - 2011
N2 - Human spermatozoal RNA features special characteristics such as a significantly reduced quantity within spermatozoa compared with somatic cells is described as being devoid of ribosomal RNAs and is difficult to isolate due to a massive excess of genomic DNA in the lysates. Using a novel two-round column-based protocol for human ejaculates delivering highly purified spermatozoal RNA, we uncovered a heterogeneous, but specific banding pattern in microelectrophoresis with 28S ribosomal RNA being indicative for the amount of round cell contamination. Ejaculates with different round cell quantities and density-purified spermatozoa revealed that 18S rRNA but not 28S rRNA is inherent to a pure spermatozoal fraction. Transmission electron microscopy showed monoribosomes and polyribosomes in spermatozoal cytoplasm, while immunohistochemical results suggest the presence of proteins from small and large ribosomal subunits in retained spermatozoal cytoplasm irrespective of 28S rRNA absence.
AB - Human spermatozoal RNA features special characteristics such as a significantly reduced quantity within spermatozoa compared with somatic cells is described as being devoid of ribosomal RNAs and is difficult to isolate due to a massive excess of genomic DNA in the lysates. Using a novel two-round column-based protocol for human ejaculates delivering highly purified spermatozoal RNA, we uncovered a heterogeneous, but specific banding pattern in microelectrophoresis with 28S ribosomal RNA being indicative for the amount of round cell contamination. Ejaculates with different round cell quantities and density-purified spermatozoa revealed that 18S rRNA but not 28S rRNA is inherent to a pure spermatozoal fraction. Transmission electron microscopy showed monoribosomes and polyribosomes in spermatozoal cytoplasm, while immunohistochemical results suggest the presence of proteins from small and large ribosomal subunits in retained spermatozoal cytoplasm irrespective of 28S rRNA absence.
KW - Humans
KW - Male
KW - Immunohistochemistry
KW - Microscopy, Electron, Transmission
KW - Polymerase Chain Reaction
KW - Electrophoresis, Polyacrylamide Gel
KW - RNA, Ribosomal, 18S/chemistry/genetics
KW - RNA, Ribosomal, 28S/chemistry/genetics
KW - Ribosomes/metabolism
KW - Spermatozoa/chemistry
KW - Humans
KW - Male
KW - Immunohistochemistry
KW - Microscopy, Electron, Transmission
KW - Polymerase Chain Reaction
KW - Electrophoresis, Polyacrylamide Gel
KW - RNA, Ribosomal, 18S/chemistry/genetics
KW - RNA, Ribosomal, 28S/chemistry/genetics
KW - Ribosomes/metabolism
KW - Spermatozoa/chemistry
M3 - SCORING: Journal article
VL - 17
SP - 669
EP - 678
JO - MOL HUM REPROD
JF - MOL HUM REPROD
SN - 1360-9947
IS - 11
M1 - 11
ER -
