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Microinjection manipulation resulted in the increased apoptosis of spermatocytes in testes from intracytoplasmic sperm injection (ICSI) derived mice.

Yu Y, Zhao C, Lv Z, Chen W, Tong M, Guo X, Wang L, Liu J, Zhou Z, Zhu H, Zhou Q, Sha J - PLoS ONE (2011)

Bottom Line: However, concerns remain regarding the safety of this method since it is a more invasive procedure than in vitro fertilization (IVF), since a spermatozoon is injected into the oocyte cytoplasm.And, decreased testis weight and marked damage of spermatogenic epithelia were found in aged ICSI mice.The changed expression of a series of proteins relating to apoptosis or proliferation might contribute to it.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, the Chinese Academy of Science, Beijing, China.

ABSTRACT
The invention of intracytoplasmic sperm injection (ICSI) has possibly been the most important development in reproductive medicine, one that has given hope to thousands of infertile couples worldwide. However, concerns remain regarding the safety of this method since it is a more invasive procedure than in vitro fertilization (IVF), since a spermatozoon is injected into the oocyte cytoplasm. Using mice derived from IVF technology as a control, we assessed the influence of invasive microinjection in the process of transferring sperm into oocyte cytoplasm in ICSI procedure on the development and physiologic function of resultant offspring. Our results demonstrated that mice produced from ICSI and IVF had no significant difference in phenotypic indices including body weight, forelimb physiology, and learning and memory ability. However, increased spermatocyte apoptosis was observed in the testis of adult ICSI mice, when compared with IVF mice. And, decreased testis weight and marked damage of spermatogenic epithelia were found in aged ICSI mice. Furthermore, proteomic analysis verified that most of the differentiated proteins in testes between adult ICSI and IVF mice were those involved in regulation of apoptosis pathways. Our results demonstrated that the microinjection manipulation used in the ICSI procedure might pose potential risks to the fertility of male offspring. The changed expression of a series of proteins relating to apoptosis or proliferation might contribute to it. Further studies are necessary to better understand all the risks of ICSI.

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Testicular morphology of aged offspring derived from ICSI and IVF.The morphology of ICSI (A) and IVF testes (B) was examined by hematoxylin and eosin (HE) staining. The ICSI testis contained much more abnormal seminiferous tubule (*) than the IVF group. The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes (C) when compared to the IVF mice (D). (E) The percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (* P<0.01). (F) The percentage of apoptotic spermatogenic cells in ICSI testes was comparable to the IVF group (P>0.05). Bar: 50 µm.
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pone-0022172-g004: Testicular morphology of aged offspring derived from ICSI and IVF.The morphology of ICSI (A) and IVF testes (B) was examined by hematoxylin and eosin (HE) staining. The ICSI testis contained much more abnormal seminiferous tubule (*) than the IVF group. The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes (C) when compared to the IVF mice (D). (E) The percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (* P<0.01). (F) The percentage of apoptotic spermatogenic cells in ICSI testes was comparable to the IVF group (P>0.05). Bar: 50 µm.

Mentions: Furthermore, histological examination of testes from aged mice was performed. We found that the morphology of seminiferous tubules from IVF mice (n = 3) appeared normal (Fig. 4B). However, a large number of abnormal seminiferous tubules, including apparent premature release of germ cells into the seminiferous tubule lumen, vacuolization in the seminiferous tubules, and germ cell-deficient (e.g., Sertoli cell only) seminiferous tubules, etc. were observed in aged ICSI testes (n = 6, Fig. 4A). And, the percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (P<0.01,Fig. 4E). The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes when compared to the IVF mice (Fig. 4C and D). However, statistical analysis showed the difference was not significant (Fig. 4F, P>0.05), probably because numerous spermatogenic cells had died in the aged ICSI mice.


Microinjection manipulation resulted in the increased apoptosis of spermatocytes in testes from intracytoplasmic sperm injection (ICSI) derived mice.

Yu Y, Zhao C, Lv Z, Chen W, Tong M, Guo X, Wang L, Liu J, Zhou Z, Zhu H, Zhou Q, Sha J - PLoS ONE (2011)

Testicular morphology of aged offspring derived from ICSI and IVF.The morphology of ICSI (A) and IVF testes (B) was examined by hematoxylin and eosin (HE) staining. The ICSI testis contained much more abnormal seminiferous tubule (*) than the IVF group. The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes (C) when compared to the IVF mice (D). (E) The percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (* P<0.01). (F) The percentage of apoptotic spermatogenic cells in ICSI testes was comparable to the IVF group (P>0.05). Bar: 50 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3140508&req=5

pone-0022172-g004: Testicular morphology of aged offspring derived from ICSI and IVF.The morphology of ICSI (A) and IVF testes (B) was examined by hematoxylin and eosin (HE) staining. The ICSI testis contained much more abnormal seminiferous tubule (*) than the IVF group. The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes (C) when compared to the IVF mice (D). (E) The percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (* P<0.01). (F) The percentage of apoptotic spermatogenic cells in ICSI testes was comparable to the IVF group (P>0.05). Bar: 50 µm.
Mentions: Furthermore, histological examination of testes from aged mice was performed. We found that the morphology of seminiferous tubules from IVF mice (n = 3) appeared normal (Fig. 4B). However, a large number of abnormal seminiferous tubules, including apparent premature release of germ cells into the seminiferous tubule lumen, vacuolization in the seminiferous tubules, and germ cell-deficient (e.g., Sertoli cell only) seminiferous tubules, etc. were observed in aged ICSI testes (n = 6, Fig. 4A). And, the percentage of seminiferous tubules with abnormal morphololgy in ICSI testes was significantly higher than that in IVF testes (P<0.01,Fig. 4E). The TUNEL assay showed more spermatogenic cells undergoing apoptosis in aged ICSI testes when compared to the IVF mice (Fig. 4C and D). However, statistical analysis showed the difference was not significant (Fig. 4F, P>0.05), probably because numerous spermatogenic cells had died in the aged ICSI mice.

Bottom Line: However, concerns remain regarding the safety of this method since it is a more invasive procedure than in vitro fertilization (IVF), since a spermatozoon is injected into the oocyte cytoplasm.And, decreased testis weight and marked damage of spermatogenic epithelia were found in aged ICSI mice.The changed expression of a series of proteins relating to apoptosis or proliferation might contribute to it.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, the Chinese Academy of Science, Beijing, China.

ABSTRACT
The invention of intracytoplasmic sperm injection (ICSI) has possibly been the most important development in reproductive medicine, one that has given hope to thousands of infertile couples worldwide. However, concerns remain regarding the safety of this method since it is a more invasive procedure than in vitro fertilization (IVF), since a spermatozoon is injected into the oocyte cytoplasm. Using mice derived from IVF technology as a control, we assessed the influence of invasive microinjection in the process of transferring sperm into oocyte cytoplasm in ICSI procedure on the development and physiologic function of resultant offspring. Our results demonstrated that mice produced from ICSI and IVF had no significant difference in phenotypic indices including body weight, forelimb physiology, and learning and memory ability. However, increased spermatocyte apoptosis was observed in the testis of adult ICSI mice, when compared with IVF mice. And, decreased testis weight and marked damage of spermatogenic epithelia were found in aged ICSI mice. Furthermore, proteomic analysis verified that most of the differentiated proteins in testes between adult ICSI and IVF mice were those involved in regulation of apoptosis pathways. Our results demonstrated that the microinjection manipulation used in the ICSI procedure might pose potential risks to the fertility of male offspring. The changed expression of a series of proteins relating to apoptosis or proliferation might contribute to it. Further studies are necessary to better understand all the risks of ICSI.

Show MeSH
Related in: MedlinePlus