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Bhilawadikar R.,Jaslok Hospital | Bhilawadikar R.,Inkus Center | Zaveri K.,Inkus Center | Mukadam L.,Inkus Center | And 5 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2013

Purpose: To compare the expression profiles of Tektin 2 and CatSper 2 motility proteins in the spermatozoa of normozoospermic and oligoasthenozoospermic men and determine its correlation with sperm motility, fertilization rate, embryo quality and pregnancy rate. Methods: Tektin 2 and CatSper 2 protein expression was studied using Western Blotting and immunofluorescence. Tektin 2 and CatSper 2 protein levels were quantified by ELISA. Results: Oligoasthenozoospermic men were found to have lower fertilization rates, poor embryo quality and lower pregnancy rates as compared to normozoospermic men. The levels of Tektin 2 and CatSper 2 are significantly lower in spermatozoa of oligoasthenozoospermic men as compared to normozoospermic controls; the levels were also lower in immotile fraction as compared to motile fraction of spermatozoa obtained from normozoospermic individuals. The levels of Tektin 2 and CatSper 2 were higher in individuals demonstrating sperm motility >60 % as compared to sperm motility <30 %. Tektin 2 but not CatSper 2 levels were positively associated with fertilization rate, embryo quality and pregnancy rate. Conclusion: Levels of Tektin 2 and CatSper 2 proteins are positively associated with sperm motility parameters. Measurements of Tektin 2 levels can be correlated with the clinical outcome of ICSI. © 2013 Springer Science+Business Media New York.


Nimbkar-Joshi S.,National Health Research Institute | Katkam R.R.,National Health Research Institute | Chaudhari U.K.,National Health Research Institute | Jacob S.,National Health Research Institute | And 8 more authors.
Histochemistry and Cell Biology | Year: 2012

The present investigation reports embryoinduced modifications in the epithelial cells of the endometrium in a primate species. In vivo, epithelial cell response to the embryonic signals was assessed at the embryo attachment stage in the gestational uterus of bonnet monkeys (Macaca radiata) and in vitro response was investigated by treating human endometrial epithelial cell line (Ishikawa) with human embryo conditioned media (CM). Endometrial epithelial (EE) cells at the embryo attachment stage in bonnet monkeys revealed higher proliferation accompanied by significant up regulation (p < 0.05) in the expression of estrogen receptor (ER)a and down regulation (p < 0.05) in ERβ expression. Further gestational EE cells showed higher (p< 0.001) expression of mucin-1, except in the embryo attachment site. Also, observed were significantly higher expression (p < 0.05) and altered cytoplasmic distribution of αv and b3 integrins, when compared to non-pregnant animals. In pregnant animals, the embryo attachment zone showed differential expression of immunoreactive integrins as compared to the non-attachment zone. This suggested the role of embryo secreted factors in modulation of the epithelial cell profile. In vitro studies partially supported this assumption. Significantly higher proliferation (p < 0.05), as well as increased expression of ERα, integrin β3 and mucin-1 (p < 0.05) were observed in Ishikawa cells, on stimulation with CM. Taken together, these results indicated the proliferation and modulation in the expression of estrogen receptors and cell adhesion molecules in the EE cells; at the embryo attachment stage in bonnet monkeys. Further it is likely that embryo secreted factors contribute to some of these modifications in EE cells. This report is the first account of discrete cellular events, which occur in the uterine epithelium, at the embryo attachment stage in a primate species. © Springer-Verlag 2012.


Nagvenkar P.,National Health Research Institute | Pethe P.,National Health Research Institute | Pawani H.,National Health Research Institute | Telang J.,National Health Research Institute | And 4 more authors.
In Vitro Cellular and Developmental Biology - Animal | Year: 2011

Human embryonic stem (hES) cells possess the ability to self-renew indefinitely and provide a potential source of differentiated progeny representing all three embryonic germ layers. Although hES cell lines share the expression of typical pluripotency markers, limited data is available regarding their differentiation capabilities. We have earlier reported the in-house derivation of two hES cell lines, KIND-1 and KIND-2 on human feeders. Here, we describe a comparative study carried out on both these cell lines to better understand the differentiation potential of KIND-1 and KIND-2 by gene expression analysis of representative gene transcripts reflecting pluripotency and the three germ layers viz. ectoderm, mesoderm, and endoderm. Gene expression analysis and immunolocalization studies were undertaken on (a) 7- and 14-d old embryoid bodies (EBs) (b) spontaneously differentiated cells from EBs, (c) cells derived from EBs under the influence of various growth factor treatments and (d) KIND-1 and KIND-2 cells co-cultured on mouse embryonic visceral endoderm-like feeder (END-2). Despite both the cell lines being XX, derived, passaged, and cultured similarly, KIND-1 exhibits preferential differentiation towards endodermal lineage whereas KIND-2 spontaneously forms beating cardiomyocytes. Perhaps the occurrence of discrete epigenetic profile in both the cell lines predisposes them to encompass different developmental potential in vitro. Our data provide evidence for existence of distinct differentiation propensity among hES cell lines and emphasizes the need to derive more hES cell lines for future regenerative medicine. © 2011 The Society for In Vitro Biology.


Hinduja I.,Jaslok Hospital and Research Center | Hinduja I.,Inkus Center | Baliga N.B.,Jaslok Hospital and Research Center | Zaveri K.,Inkus Center
Journal of Human Reproductive Sciences | Year: 2010

Objective: The centrosome is the microtubule organizing center (MTOC) paternally inherited by the zygote during fertilization. As the centrosome is located in the midpiece of the sperm tail, we presume that oligoasthenozoospermic sperm samples should also have abnormal concentrations of centrosomal proteins. This study therefore aims to determine if there is any correlation between sperm centrosomal proteins, centrin, α and γ-tubulin, in sperm samples from normozoospermic and oligoasthenozoospermic men. Materials and Methods: Proteins were extracted from the normozoospermic and oligoasthenozoospermic sperm samples and analyzed by Western Blot and ELISA for centrin, α and γ -tubulin. Results: The levels of centrin, α and γ-tubulin are markedly lower in oligoasthenozoospermic sperm samples as compared to the normozoospermic sperm samples. Conclusions: Lower centrosomal protein expression in sperm samples of oligoasthenozoospermic infertile males may be a possible cause for their reduced fertility status. Further studies on these proteins are warranted to design rational approaches for the diagnosis and treatment of male infertility.

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