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. 2009 Apr;1(1):41-6.

Assessment of thyroglobulin expression in reproductive organs at different stages of mouse estrous cycle

Ali Moravvej  1 , Mahmood Jeddi-Tehrani, Ali Reza Salek Moghaddam, Pouneh Dokouhaki, Mahdi Shekarabi, Roya Ghods, Mahdi Shahbazi, Jamileh Ghasemi, Parivash Danesh, Ahmad Reza Mahmoudi, Amir Hassan Zarnani
Affiliations

Assessment of thyroglobulin expression in reproductive organs at different stages of mouse estrous cycle

Ali Moravvej et al. Avicenna J Med Biotechnol. 2009 Apr.

Abstract

Prevalence of abortion is higher in women with autoimmune thyroid disease. In the majority of cases, however, no abnormality of thyroid function is detected despite the high levels of antithyroid antibodies. The direct influence of such harmful autoantibodies in female reproductive organs may serve a role in pregnancy loss. In this study, expression of thyroglobulin in the reproductive tissues of cycling mice has been evaluated. Stages of estrous cycle were determined by cellular morphology and ratio of epithelial cells to leukocytes in vaginal smear of Balb/C mice. At each phase, the mice were sacrificed and their uterus, ovary and fallopian tubes were removed. Expression of thyroglobulin-specific transcript in endometrium was investigated by two sets of primers using reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, expression of thyroglobulin in reproductive tissues was assessed by immunohistochemistry and dot blot analysis. The results showed that thyroglobulin mRNA is not expressed in endometrial tissue of Balb/C mice at any stage of estrous cycle. Immunohistochemical analysis also confirmed that thyroglobulin or its cross reactive-antigens are not expressed at the protein level in the female reproductive organs. The results showed that thyroglobulin was not expressed in the reproductive organs of female mice. It is plausible that antithyroglobulin antibodies could interact with newly-generated antigens during placentation and pregnancy.

Keywords: Antithyroglobulin antibody; Endometrium; Estrous cycle; Placentation; Recurrent abortion; Uterus.

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Figures

Figure 1
Figure 1
Amplification of TG gene in endometrium of cycling mice by RT-PCR Endometrium of cycling mice was pealed away during each stage of estrous cycle and expression of TG gene was assessed by two sets of primers (TG1 & TG2). GAPDH amplification was used as internal control for RNA extraction and cDNA synthesis. 309bp band of amplified GAPDH mRNA was found in all endometrial and thyroid samples. On the contrary, 207bp band corresponding to the amplified segments of TG1 (a) or 510bp band corresponding to the amplified segments of TG2 (b) were not detected in any endometrial samples but it was strongly detected in thyroid tissue.
Figure 2
Figure 2
Immunostaining of mouse thyroid with polyclonal rabbit antiTG antibody To confirm reactivity of homemade antiTG antibody, immunohistochemistry staining of frozen (a) and paraffin embedded (c) sections of mouse thyroid was performed. In both methods the antibody showed strong immonoreactivity. b and d: Negative control slides of frozen and paraffin embedded sections, respectively. (Magnification: 200 ×).
Figure 3
Figure 3
Immunohistochemical analysis of reproductive organs of cycling mice with antiTG antibody Uterus, ovary and fallopian tubes of cycling Balb/C mice were tested by immunohistochemistry for expression of TG. TG was not expressed in any of the tissues at all stages of estrous cycle. a-d: endometrium in proestrous, estrus, metestrous and diestrous, respectively. e and f: representative of ovary and fallopian tube. g-i: negative controls of endometrium, ovary and fallopian tubes. j: thyroid as positive control.
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