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Hum. Reprod. Advance Access published online on May 21, 2009
Human Reproduction, doi:10.1093/humrep/dep187
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© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Differential expression of calreticulin, a reticuloplasmin in primate endometrium

T. Parmar1, S. Nimbkar-Joshi1, R.R. Katkam1, S. Gadkar-Sable1, U. Chaudhari1, D.D. Manjramkar2, L. Savardekar3, S. Jacob1, C.P. Puri1 and G. Sachdeva1,4

1 Primate Biology Division, National Institute for Research in Reproductive Health, Indian Council of Medical Research, Parel, Mumbai 400012, India 2 Experimental Animal Facilities, National Institute for Research in Reproductive Health, Indian Council of Medical Research, Parel, Mumbai 400012, India 3 Clinical Research Division, National Institute for Research in Reproductive Health, Indian Council of Medical Research, Parel, Mumbai 400012, India

4 Correspondence address. Tel: +91-22-24192111; Fax: +91-22-24139412; E-mail: sachdevag{at}nirrh.res.in

BACKGROUND: To our knowledge, there are no data on hormonal regulation of reticuloplasmins in primate endometrium. We report the presence and modulation of expression of three reticuloplasmins in endometrium of bonnet monkeys (Macaca radiata).

METHODS: Receptive and non-receptive endometria obtained from vehicle-treated control and onapristone (antiprogestin)-treated animals, respectively, were compared for differentially expressed proteins by two-dimensional proteomics. Mass spectrometric analysis annotated two such proteins as calreticulin and protein disulfide-isomerase (PDI), known to be molecular chaperones in endoplasmic reticulum. We then investigated if endoplasmin, another reticuloplasmin is also differentially expressed. Expression of these reticuloplasmins was also investigated in the endometriuma during pregnancy in bonnet monkeys. Samples were analysed by immunohistochemistry and western blot (calreticulin in human endometrium), and calreticulin transcript levels in Ishikawa cell line were assessed by real time PCR.

RESULTS: Immunohistochemical analysis of the functionalis region of non-receptive endometria in monkeys revealed higher expression of (i) calreticulin (P < 0.01) in glandular epithelium and (ii) PDI in stroma (P < 0.0001), but no change in endoplasmin in stroma or glands, compared with receptive endometria. Protein level of all three reticuloplasmins in the stromal region of endometrial functionalis was higher in pregnant than non-pregnant animals (P < 0.05). Human endometrial calreticulin protein was higher in the estrogen-dominant (proliferative) phase than progesterone-dominant (mid-secretory) phase of the cycle. Calreticulin mRNA in Ishikawa cells is up-regulated by estrogen (P < 0.05 versus control), with a trend towards down-regulation by progesterone.

CONCLUSION: Our data suggest that endometrial reticuloplasmins are regulated by hormones and embryonic stimuli in a cell-type specific manner. These novel data open up new lines of investigation for elucidating the mechanisms by which hormones or embryonic stimuli influence the sub-cellular physiology of endometrium.

Key words: endometrium/receptivity/reticuloplasmins/calreticulin/protein disulfide-isomerase

Submitted on January 14, 2009; resubmitted on April 16, 2009; accepted on April 21, 2009.


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